FU-tet-o-hOct4人源基因质粒

基本信息

启动子:	minimal CMV promoter
复制子:	pUC ori,F1 ori,SV40 ori
终止子:	WPRE
质粒分类:	基因文库质粒；人源基因质粒；人源真核质粒
质粒大小:	9471bp
原核抗性:	Amp（100μg/ml）
筛选标记:	Zeo
克隆菌株:	Stbl3
培养条件:	37℃，LB，有氧
表达宿主:	293T等哺乳细胞
培养条件:	37℃，5%CO2
诱导方式:	四环素调控
5'测序引物:	miniCMV-F（GCGTGTACGGTGGGAGGCC）
3'测序引物:	WPRE-R（CATAGCGTAAAAGGAGCAACA）

质粒简介
             pFU-tet-o-hOct4质粒是一个人源Oct4基因的哺乳细胞慢病毒过表达质粒，minimal CMV promoter启动Oct4目的基因的表达。该质粒可作为过表达质粒直接转染哺乳细胞，也可包装成慢病毒侵染细胞。质粒本身带有Zeo筛选标记，可用博来霉素筛选转染后的细胞得到稳转细胞系。基因NCBI信息：Homo sapiens POU class 5 homeobox 1 (POU5F1), transcript variant 1, mRNA；NCBI Reference Sequence: NM_002701.5.质粒图谱

质粒序列
LOCUS       Exported                9471 bp ds-DNA    circular SYN 11-七月-2017
KEYWORDS    pFU-tet-o-hOct4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9471)
 TITLE     Direct Submission
  JOURNAL   Exported 2017年7月11日 from Plasmid
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9471
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        238..617
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             835..1015
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /note="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1680..1913
                     /note="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     misc_feature    2440..2557
                     /note="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     protein_bind    2610..2880
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tetracycline response element"
                     /note="contains seven copies of the tetracycline operator
                     tetO"
     promoter        2913..2950
                     /note="minimal CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             3056..4138
                     /codon_start=1
                     /product="Homo sapiens POU class 5 homeobox 1 (POU5F1),
                     transcript variant 1, mRNA
                     NCBI Reference Sequence: NM_002701.5"
                     /note="Oct4"
                     /translation="MAGHLASDFAFSPPPGGGGDGPGGPEPGWVDPRTWLSFQGPPGGP
                     GIGPGVGPGSEVWGIPPCPPPYEFCGGMAYCGPQVGVGLVPQGGLETSQPEGEAGVGVE
                     SNSDGASPEPCTVTPGAVKLEKEKLEQNPEESQDIKALQKELEQFAKLLKQKRITLGYT
                     QADVGLTLGVLFGKVFSQTTICRFEALQLSFKNMCKLRPLLQKWVEEADNNENLQEICK
                     AETLVQARKRKRTSIENRVRGNLENLFLQCPKPTLQQISHIAQQLGLEKDVVRVWFCNR
                     RQKGKRSSSDYAQREDFEAAGSPFSGGPVSFPLAPGPHFGTPGYGSPHFTALYSSVPFP
                     EGEAFPPVSVTTLGSPMHSN"
     misc_feature    4163..4751
                     /note="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5276..5456
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    5488..5712
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5758..6186
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6200..6529
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      6380..6515
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        6577..6624
                     /note="EM7 promoter"
                     /note="synthetic bacterial promoter "
     CDS             6643..7017
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /note="Zeo"
                     /note="confers resistance to bleomycin, phleomycin, and
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    7147..7268
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(7719..8307)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8478..9338)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9339..9443)
                     /gene="bla"
                     /note="AmpR promoter"
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttatcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc
     2641 gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat
     2701 cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg
     2761 aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact
     2821 ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga
     2881 aaagtgaaag tcgagctcgg tacccgggtc gagtaggcgt gtacggtggg aggcctatat
     2941 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
     3001 cctccataga agacaccggg accgatccag cctccgcggc cccgaattcg ccaccatggc
     3061 gggacacctg gcttcggatt tcgccttctc gccccctcca ggtggtggag gtgatgggcc
     3121 aggggggccg gagccgggct gggttgatcc tcggacctgg ctaagcttcc aaggccctcc
     3181 tggagggcca ggaatcgggc cgggggttgg gccaggctct gaggtgtggg ggattccccc
     3241 atgccccccg ccgtatgagt tctgtggggg gatggcgtac tgtgggcccc aggttggagt
     3301 ggggctagtg ccccaaggcg gcttggagac ctctcagcct gagggcgaag caggagtcgg
     3361 ggtggagagc aactccgatg gggcctcccc ggagccctgc accgtcaccc ctggtgccgt
     3421 gaagctggag aaggagaagc tggagcaaaa cccggaggag tcccaggaca tcaaagctct
     3481 gcagaaagaa ctcgagcaat ttgccaagct cctgaagcag aagaggatca ccctgggata
     3541 tacacaggcc gatgtggggc tcaccctggg ggttctattt gggaaggtat tcagccaaac
     3601 gaccatctgc cgctttgagg ctctgcagct tagcttcaag aacatgtgta agctgcggcc
     3661 cttgctgcag aagtgggtgg aggaagctga caacaatgaa aatcttcagg agatatgcaa
     3721 agcagaaacc ctcgtgcagg cccgaaagag aaagcgaacc agtatcgaga accgagtgag
     3781 aggcaacctg gagaatttgt tcctgcagtg cccgaaaccc acactgcagc agatcagcca
     3841 catcgcccag cagcttgggc tcgagaagga tgtggtccga gtgtggttct gtaaccggcg
     3901 ccagaagggc aagcgatcaa gcagcgacta tgcacaacga gaggattttg aggctgctgg
     3961 gtctcctttc tcagggggac cagtgtcctt tcctctggcc ccagggcccc attttggtac
     4021 cccaggctat gggagccctc acttcactgc actgtactcc tcggtccctt tccctgaggg
     4081 ggaagccttt ccccctgtct ctgtcaccac tctgggctct cccatgcatt caaactgaga
     4141 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     4201 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     4261 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     4321 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     4381 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     4441 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4501 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     4561 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4621 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4681 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4741 cgcctccccg catcgatacc gtcgacctcg agacctagaa aaacatggag caatcacaag
     4801 tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga
     4861 ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt
     4921 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     4981 aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattggca
     5041 gaactacaca ccagggccag ggatcagata tccactgacc tttggatggt gctacaagct
     5101 agtaccagtt gagcaagaga aggtagaaga agccaatgaa ggagagaaca cccgcttgtt
     5161 acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtattag agtggaggtt
     5221 tgacagccgc ctagcatttc atcacatggc ccgagagctg catccggact gtactgggtc
     5281 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     5341 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     5401 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagggc
     5461 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     5521 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     5581 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     5641 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     5701 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     5761 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5821 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     5881 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5941 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     6001 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     6061 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     6121 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     6181 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     6241 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     6301 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     6361 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     6421 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     6481 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     6541 agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag
     6601 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt
     6661 gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg
     6721 ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg
     6781 accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg
     6841 tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc
     6901 cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc
     6961 gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac
     7021 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     7081 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     7141 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     7201 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     7261 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     7321 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     7381 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     7441 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     7501 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     7561 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     7621 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     7681 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7741 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7801 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7861 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7921 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7981 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     8041 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     8101 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     8161 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     8221 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     8281 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8341 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8401 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8461 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8521 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8581 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8641 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8701 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8761 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8821 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8881 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8941 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     9001 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     9061 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     9121 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     9181 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     9241 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9301 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9361 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9421 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga c