pECMV- 3脳 FLAG- AFP/ pECMV- 3脳 FLAG- AFP质粒/ pECMV- 3脳 FLAG- AFP Plasmid

pECMV-3×FLAG-AFP

 

Alternative Name

pECMV-3×FLAG-AFP, pECMV-3×FLAG-AFP plasmid, pECMV-3×FLAG-AFP Vector

 

pECMV-3×FLAG-AFP Information

Prokaryotic resistance: ampicillin Amp

 

Screening markers: neomycin Neo/ hereditary mycin G418

 

Cloned strain: Escherichia coli DH5 alpha

 

Culture conditions: 37 degrees centigrade

 

 

 

pECMV-3×FLAG-AFP Description

pECMV-3×FLAG-AFP is a human mammalian expression plasmid

 

pECMV-3×FLAG-AFP Sequence

LOCUS       Exported                7282 bp ds-DNA     circular SYN 28-SEP-2017

DEFINITION  synthetic circular DNA

  TITLE     Direct Submission

  JOURNAL   Exported Sep 28, 2017  

FEATURES             Location/Qualifiers

     source          1..7282

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     enhancer        235..614

                     /label=CMV enhancer

                     /note="human cytomegalovirus immediate early enhancer"

     promoter        615..818

                     /label=CMV promoter

                     /note="human cytomegalovirus (CMV) immediate early 

                     promoter"

     promoter        863..881

                     /label=T7 promoter

                     /note="promoter for bacteriophage T7 RNA polymerase"

     CDS             922..987

                     /codon_start=1

                     /product="three tandem FLAG(R) epitope tags, followed by an

                     enterokinase cleavage site"

                     /label=3xFLAG

                     /translation="DYKDHDGDYKDHDIDYKDDDDK"

     misc_feature    1009..2838

                     /label=AFP

     polyA_signal    2882..3106

                     /label=bGH poly(A) signal

                     /note="bovine growth hormone polyadenylation signal"

     rep_origin      3152..3580

                     /direction=RIGHT

                     /label=f1 ori

                     /note="f1 bacteriophage origin of replication; arrow 

                     indicates direction of (+) strand synthesis"

     promoter        3594..3923

                     /label=SV40 promoter

                     /note="SV40 enhancer and early promoter"

     rep_origin      3774..3909

                     /label=SV40 ori

                     /note="SV40 origin of replication"

     CDS             3990..4784

                     /codon_start=1

                     /gene="aph(3')-II (or nptII)"

                     /product="aminoglycoside phosphotransferase from Tn5"

                     /label=Neo

                     /note="confers resistance to neomycin, kanamycin, and G418 

                     (Geneticin(R))"

                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP

                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS

                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ

                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA

                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"

     polyA_signal    4958..5079

                     /label=SV40 poly(A) signal

                     /note="SV40 polyadenylation signal"

     rep_origin      complement(5530..6115)

                     /direction=LEFT

                     /label=ori

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 

                     replication"

     CDS             complement(6286..7146)

                     /codon_start=1

                     /gene="bla"

                     /product="beta-lactamase"

                     /label=AmpR

                     /note="confers resistance to ampicillin, carbenicillin, and

                     related antibiotics"

                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI

                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS

                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW

                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA

                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS

                     LIKHW"

     promoter        complement(7147..7251)

                     /gene="bla"

                     /label=AmpR promoter

ORIGIN

        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg

       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg

      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc

      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt

      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata

      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc

      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc

      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt

      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt

      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca

      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg

      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc

      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg

      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca

      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc

      901 gtttaaactt aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac

      961 atcgactaca aggatgacga tgacaagctt ggtaccgagc tcggatccat gaagtgggtg

     1021 gaatcaattt ttttaatttt cctactaaat tttactgaat ccagaacact gcatagaaat

     1081 gaatatggaa tagcttccat attggattct taccaatgta ctgcagagat aagtttagct

     1141 gacctggcta ccatattttt tgcccagttt gttcaagaag ccacttacaa ggaagtaagc

     1201 aaaatggtga aagatgcatt gactgcaatt gagaaaccca ctggagatga acagtcttca

     1261 gggtgtttag aaaaccagct acctgccttt ctggaagaac tttgccatga gaaagaaatt

     1321 ttggagaagt acggacattc agactgctgc agccaaagtg aagagggaag acataactgt

     1381 tttcttgcac acaaaaagcc cactccagca tcgatcccac ttttccaagt tccagaacct

     1441 gtcacaagct gtgaagcata tgaagaagac agggagacat tcatgaacaa attcatttat

     1501 gagatagcaa gaaggcatcc cttcctgtat gcacctacaa ttcttctttg ggctgctcgc

     1561 tatgacaaaa taattccatc ttgctgcaaa gctgaaaatg cagttgaatg cttccaaaca

     1621 aaggcagcaa cagttacaaa agaattaaga gaaagcagct tgttaaatca acatgcatgt

     1681 gcagtaatga aaaattttgg gacccgaact ttccaagcca taactgttac taaactgagt

     1741 cagaagttta ccaaagttaa ttttactgaa atccagaaac tagtcctgga tgtggcccat

     1801 gtacatgagc actgttgcag aggagatgtg ctggattgtc tgcaggatgg ggaaaaaatc

     1861 atgtcctaca tatgttctca acaagacact ctgtcaaaca aaataacaga atgctgcaaa

     1921 ctgaccacgc tggaacgtgg tcaatgtata attcatgcag aaaatgatga aaaacctgaa

     1981 ggtctatctc caaatctaaa caggttttta ggagatagag attttaacca attttcttca

     2041 ggggaaaaaa atatcttctt ggcaagtttt gttcatgaat attcaagaag acatcctcag

     2101 cttgctgtct cagtaattct aagagttgct aaaggatacc aggagttatt ggagaagtgt

     2161 ttccagactg aaaaccctct tgaatgccaa gataaaggag aagaagaatt acagaaatac

     2221 atccaggaga gccaagcatt ggcaaagcga agctgcggcc tcttccagaa actaggagaa

     2281 tattacttac aaaatgcgtt tctcgttgct tacacaaaga aagcccccca gctgacctcg

     2341 tcggagctga tggccatcac cagaaaaatg gcagccacag cagccacttg ttgccaactc

     2401 agtgaggaca aactattggc ctgtggcgag ggagcggctg acattattat cggacactta

     2461 tgtatcagac atgaaatgac tccagtaaac cctggtgttg gccagtgctg cacttcttca

     2521 tatgccaaca ggaggccatg cttcagcagc ttggtggtgg atgaaacata tgtccctcct

     2581 gcattctctg atgacaagtt cattttccat aaggatctgt gccaagctca gggtgtagcg

     2641 ctgcaaacga tgaagcaaga gtttctcatt aaccttgtga agcaaaagcc acaaataaca

     2701 gaggaacaac ttgaggctgt cattgcagat ttctcaggcc tgttggagaa atgctgccaa

     2761 ggccaggaac aggaagtctg ctttgctgaa gagggacaaa aactgatttc aaaaactcgt

     2821 gctgctttgg gagtttaact cgagtctaga gggcccgttt aaacccgctg atcagcctcg

     2881 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc

     2941 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt

     3001 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat

     3061 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa

     3121 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg

     3181 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct

     3241 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta

     3301 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa

     3361 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct

     3421 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc

     3481 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg

     3541 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc

     3601 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc

     3661 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc

     3721 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc

     3781 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt

     3841 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt

     3901 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg

     3961 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt

     4021 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct

     4081 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga

     4141 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg

     4201 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact

     4261 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg

     4321 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct

     4381 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg

     4441 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt

     4501 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg

     4561 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc

     4621 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag

     4681 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt

     4741 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt

     4801 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc

     4861 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca

     4921 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa

     4981 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca

     5041 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac

     5101 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc

     5161 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta

     5221 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa

     5281 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat

     5341 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg

     5401 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc

     5461 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt

     5521 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag

     5581 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc

     5641 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc

     5701 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt

     5761 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt

     5821 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc

     5881 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa

     5941 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa

     6001 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg

     6061 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga

     6121 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat

     6181 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag

     6241 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat

     6301 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc

     6361 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat

     6421 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag

     6481 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg

     6541 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc

     6601 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca

     6661 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg

     6721 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc

     6781 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta

     6841 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc

     6901 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg

     6961 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc

     7021 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc

     7081 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat

     7141 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag

     7201 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc

     7261 ccgaaaagtg ccacctgacg tc

//

 

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