基本信息
| 别名: | MDC1,NFBD1,KIAA0170 |
| 复制子: | pUC ori |
| 质粒大小: | 9539bp |
| 质粒标签: | N-EGFP |
| 原核抗性: | 卡那霉素Kan |
| 克隆菌株: | 大肠杆菌DH5α |
| 培养条件: | 37℃，有氧，LB |
质粒简介
pENTR-GFP-MDC1 (770-7)质粒是一个携带人源MDC1基因的质粒，并在N端融合了EGFP绿色荧光蛋白基因。该质粒可通过GateWay方法将EGFP-MDC1基因片段导入到其它GateWay表达载体中。质粒图谱
质粒序列
LOCUS Exported 9539 bp ds-DNA circular SYN 26-OCT-2017
DEFINITION synthetic circular DNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9539)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..9539
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 103..189
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 281..308
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
protein_bind 358..457
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 498..507
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 504..1220
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
misc_feature 1257..7556
/label=MDC1
/note="Gene/Insert name Mediator of DNA damage checkpoint 1
Alt name MDC1
Alt name NFBD1
Alt name KIAA0170
Species H. sapiens (human)
Insert Size (bp) 6269
Entrez Gene MDC1 (a.k.a. NFBD1)
Tag / Fusion Protein
GFP (N terminal on backbone)"
protein_bind complement(7768..7867)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS 7990..8799
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 8889..9477
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
421 ttttttataa tgccaacttt gtacaaaaaa gcaggctcca ccatgggaac caattcagtc
481 gactagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
541 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
601 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
661 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
721 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
781 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
841 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
901 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
961 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
1021 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
1081 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
1141 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
1201 gcatggacga gctgtacaag tccggccgga ctcagatctc gagctcaagc ttcgaattct
1261 gcagtcgacg gtaccgcggg cccatcatgg aggacaccca ggctattgac tgggatgttg
1321 aagaagagga ggagacagag caatccagtg aatccttgag gtgtaacgtg gagccagtag
1381 ggcggctaca tatctttagt ggtgcccatg gaccagaaaa agatttccca ctacacctcg
1441 ggaagaatgt ggtaggccga atgcctgact gctctgtggc cctgcccttt ccatctatct
1501 ccaaacaaca tgcagagatt gaaatcttag cctgggacaa ggcacctatc ctccgagact
1561 gtgggagcct taatggtact caaatcctga gacctcctaa ggttttgagc cctggggtga
1621 gtcaccgtct gagggaccag gaattgattc tctttgctga cttgctctgc cagtaccatc
1681 gcctggatgt ctctctgccc tttgtctccc ggggccctct gacagtagaa gagacaccca
1741 gagtacaggg agaaactcaa ccccagaggc ttctgttggc tgaggactcg gaggaggaag
1801 tagattttct ttctgaaagg cgtatggtaa aaaaatcaag gaccacatct tcctctgtga
1861 tagttccaga gagtgatgaa gaggggcatt ccccggtcct gggcggcctt gggccgcctt
1921 ttgccttcaa tttgaacagt gacacagatg tggaagaagg tcagcaacca gccacagagg
1981 aggcctcctc agctgccaga agaggtgcca ctgtagaggc aaagcagtct gaagctgaag
2041 ttgtaactga aatccagctt gaaaaggatc agcctttagt gaaggagagg gacaatgata
2101 caaaagtcaa gaggggtgca gggaatgggg tggttccagc tggggtgatt ctggagagga
2161 gccaacctcc tggagaggac agtgacacag atgtggatga tgacagcagg cctcctggaa
2221 ggccagctga ggtccatttg gaaagggctc agccttttgg cttcatcgac agcgacactg
2281 atgcggaaga agagaggatc ccagcaaccc cagttgtcat tcctatgaag aagaggaaga
2341 tcttccatgg agtaggtaca aggggtcctg gagcaccagg cctggcccat ctgcaggaga
2401 gccaggctgg tagtgataca gatgtggaag aaggcaaggc cccacaggct gtccctctgg
2461 agaaaagcca agcttccatg gttatcaaca gcgatacaga tgacgaggaa gaagtctcag
2521 cagcgctgac tttggcacat ctgaaagaga gccagcctgc tatatggaac agagatgcag
2581 aagaggacat gccccaacgt gtggtccttc tgcagcgaag ccaaaccacc actgagagag
2641 acagtgacac agacgtggag gaggaagagc tcccagtgga aaatagagaa gctgtcctca
2701 aggatcacac aaagattaga gcccttgtta gagcacattc agaaaaggac caacctcctt
2761 ttggggacag tgatgacagt gtggaagcag ataagagctc acctgggatc cacctggaga
2821 gaagccaagc ctccaccaca gtggacatca acacacaagt ggagaaggaa gtcccgccag
2881 ggtcagccat tatgcatata aagaagcatc aggtgtctgt ggaggggaca aatcaaacag
2941 atgtgaaagc agttggggga ccagcaaagc tgcttgtggt atctctagag gaagcctggc
3001 ctctgcatgg ggactgtgaa acagatgcag aggagggcac ctccctaaca gcctcagtag
3061 ttgcagatgt aagaaagagc cagcttccag cagaagggga tgctggggca gagtgggctg
3121 cagctgttct taagcaggag agagctcatg aggtgggggc ccagggtggg ccacctgtgg
3181 cacaagtgga gcaggacctc cctatctcaa gagagaacct cacagatctg gtggtggaca
3241 cagacactct aggggaatcc acccagccac agagagaggg agcccaggtc cccacaggaa
3301 gggagagaga acaacatgtg ggtgggacca aggactctga agacaactat ggtgattctg
3361 aagatctgga cctacaagct acccagtgct ttctggagaa tcagggcctg gaagcagtcc
3421 agagcatgga ggatgaacct acccaggcct tcatgttgac tccaccccaa gagcttggcc
3481 cttcccattg cagcttccag acaacaggta ccctagatga accatgggag gtcctggcta
3541 cacagccatt ctgtctgaga gagtctgagg actctgagac ccagcctttt gacacgcacc
3601 ttgaggccta tggaccttgc ctgtctccac ctagggcaat accaggagac caacatccag
3661 agagcccagt tcacacagag ccaatgggga ttcaaggcag agggaggcag actgtggata
3721 aagtcatggg tataccaaaa gaaacagcag agagggtggg ccctgagaga gggccattgg
3781 agagagaaac tgagaaactg ctaccagaaa gacagacaga tgtgacagga gaggaagaat
3841 taaccaaggg gaaacaggac agagaacaaa aacagttgtt agctagagac acccagagac
3901 aagaatctga caaaaatggg gaaagtgcaa gtcctgaaag agatagggag agtttgaagg
3961 tagaaattga gacatctgag gaaatacaag agaaacaagt acagaagcag acccttccaa
4021 gcaaagcatt tgagagagaa gtagagagac cagtagcaaa cagagagtgc gatccagccg
4081 agttagaaga gaaggtgccc aaagtgatcc tggagagaga tacacagaga ggggagccag
4141 agggagggag ccaggaccag aaagggcagg cctccagccc aacaccagag cctggggtgg
4201 gggcggggga ccttccggga cctacctcag cccccgtacc ttctgggagc cagtcaggtg
4261 gaaggggatc cccagtgagc cccaggaggc atcagaaagg cctcctgaat tgcaagatgc
4321 cacctgctga gaaggcttcc aggatcagag ctgctgagaa ggtttccagg ggcgatcagg
4381 aatctccaga tgcttgtctg cctcctgcag tacctgaagc cccagcccca ccccaaaagc
4441 cccttaactc tcagagccag aaacatcttg cacctccgcc ccttctttct ccccttttac
4501 cttctatcaa gccaaccgtt cgtaagacca ggcaagatgg gagtcaggaa gctccagagg
4561 ctcccttgtc ctcagagctg gagcctttcc acccaaagcc taaaattaga actcggaagt
4621 cctccagaat gacacccttt ccagctacct ctgctgcccc tgagccccac ccttccacct
4681 ccacagccca gccagtcact cccaagccca catctcaggc cactaggagc aggacaaata
4741 ggtcctctgt caagacccct gaaccagttg tccccacagc ccctgagctc cagccttcca
4801 cctccacaga ccagcctgtc acctctgagc ccacatctca ggttactagg ggaagaaaaa
4861 gtagatcctc tgtcaagacc cctgaaacag ttgtgcccac agcccttgag ctccagcctt
4921 ccacctccac cgaccgacct gtcacctctg aacccacctc tcaggctact aggggaagaa
4981 aaaatagatc ctctgtcaag acccctgaac cagttgtccc cacagcccct gagctccagc
5041 cttccacctc cacagaccag cctgtcactt ctgagcccac atatcaggct actaggggaa
5101 gaaaaaatag atcctctgtc aagacccctg aaccagttgt gcccacagcc cctgagctcc
5161 ggccttccac ctccacagac cgacctgtca cccccaagcc cacatctcgg accactagga
5221 gcaggacaaa tatgtcctct gtcaagaccc ctgaaacagt tgtccccaca gcccctgagc
5281 tccagatttc cacctccaca gaccaacctg tcacccctaa gcccacatct cggaccacta
5341 ggagcaggac aaatatgtcc tctgtgaaga accctgaatc aactgtccct atagcccctg
5401 agctcccacc ttccacctcc acagagcagc ctgtcacccc tgagcccaca tctcgggcta
5461 ctaggggaag aaaaaataga tcctctggca agacccctga aacacttgtc cccacagccc
5521 ctaagctcga gccttccact tccacagacc aacctgtcac tcctgagccc acatctcagg
5581 ccaccagggg caggacaaat aggtcctctg tgaagacccc tgaaacagtt gtccccacag
5641 cccctgagct ccagccttcc acctccacag accagcctgt tacccctgag cctacgtctc
5701 aggctactag gggaagaaca gatagatcct ctgtcaagac tcctgaaaca gttgtcccca
5761 cagcccctga gctacaggct tccgcctcca cagaccagcc tgtcacctct gagcccacat
5821 ctcggaccac taggggaaga aaaaatcggt cctctgtcaa gacccctgaa acagttgtgc
5881 ccgcagcccc tgagctccag cctcccacct ccacagaccg acctgtcacc cctgagccca
5941 catctcgggc cactaggggc aggacaaata ggtcctctgt caagacccct gaatcaattg
6001 tccctatagc ccctgagctt cagccttcca cctccagaaa ccagcttgtc acccctgagc
6061 ccacatctcg ggccactagg tgcaggacaa ataggtcctc tgtcaagacc cctgagccag
6121 ttgtccccac agcccctgag ccccatccta ccacctccac agaccagcct gtcaccccca
6181 agctcacatc tagggccact aggagaaaga caaataggtc ctctgtcaag actcccaaac
6241 cagttgaacc agcagcctct gatcttgagc cttttacccc cacagaccag tccgtcaccc
6301 ctgaggccat agctcagggt ggtcagagca aaacactgag gtcttccaca gtaagagcta
6361 tgccggttcc taccacccct gaattccaat ctcctgtcac cacagaccag cctatttccc
6421 ctgagcctat tactcaaccc agttgcatca agaggcagag agccgctggg aaccctggct
6481 ccctcgcagc tcccattgac cataagcctt gctctgcacc cttggaacct aaatcccagg
6541 cctcaaggaa ccaaagatgg ggagcagtga gagcagctga atcccttaca gccattcctg
6601 agcctgcctc tccccagctt cttgagacac caattcatgc ctcccagatc caaaaggtgg
6661 aaccagcagg tagatctagg ttcaccccgg agctccagcc taaggcctct caaagccgca
6721 agaggtcttt agctaccatg gattcaccac cacatcaaaa acagccccaa agaggggaag
6781 tctcccagaa gacagtgatt atcaaggaag aggaagaaga tactgcagag aagccaggga
6841 aggaagagga tgtcgtgact ccaaaaccag gcaagagaaa gagagaccag gcagaggagg
6901 agcccaacag aataccaagc cgcagcctcc gacggaccaa acttaaccaa gaatcaacag
6961 cccccaaagt gctcttcaca ggagtggtgg atgctcgggg agagcgggct gtgctggcac
7021 tggggggaag tctggctggt tcagcggcag aggcttccca cctggtcact gatcgcatcc
7081 gccggacagt caagttcctg tgtgccctgg ggcggggaat ccccattctg tccctggact
7141 ggctgcatca gtcccgcaag gctggtttct tcttaccccc ggatgaatat gtggtgaccg
7201 accctgagca agagaagaac tttggcttta gccttcaaga cgcactgagc agggctcggg
7261 agcgaaggct gctagagggc tatgagatct atgtgacccc tggagtccag ccaccaccac
7321 ctcagatggg agagattatt agctgctgtg gaggcacata cctacccagc atgcctcggt
7381 cctataagcc tcagagagtt gtgatcacat gccctcagga cttccctcat tgctccattc
7441 cactacgggt tgggctgccc ctcctctcgc ctgagttcct gctgactgga gtgctgaagc
7501 aggaagccaa gccagaggcc tttgtcctct cccctttgga gatgtcatcc acctgagaac
7561 tccactaccc ttttccctcc cagaccacga attagaagat atgtggaaga aagaactcag
7621 ggcgttagaa aggattgggg tatattgata caacttgtcc tggaacatgg gtgggaccag
7681 aaatctttat gaataaatga aaagataagg gatttggaag ccacaggttg ttttttgttt
7741 gtttgtttgt ttttttaatg gatctagacc cagctttctt gtacaaagtt ggcattataa
7801 gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca
7861 ttatttgcca tccagctgca gctctggccc gtgtctcaaa atctctgatg ttacattgca
7921 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
7981 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
8041 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
8101 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
8161 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
8221 cctcttccga ccatcaagca ttttatccgt actcctggtg atgcatggtt actcaccact
8281 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
8341 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
8401 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
8461 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
8521 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
8581 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
8641 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
8701 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
8761 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
8821 taacattatt cagattgggc cccgttccac tgagcgtcag accccgtaga aaagatcaaa
8881 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
8941 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
9001 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
9061 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
9121 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
9181 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
9241 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
9301 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
9361 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
9421 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
9481 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt
温馨提示：不可用于临床ZL。
