质粒简介
pEF-GFP是一个哺乳动物细胞中高水平表达质粒，EF1α启动子驱动目的基因和绿色荧光蛋白EGFP融合表达，MCS区域中有EcoRI、KpnI、XmaI和SmaI酶切位点，可以很方便的克隆进目的基因。
质粒序列
LOCUS Exported 5051 bp ds-DNA circular SYN 19-九月-2016
DEFINITION synthetic circular DNA
KEYWORDS Untitled 2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5051)
TITLE Direct Submission
JOURNAL Exported 2016年9月19日 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..5051
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 36..1217
/note="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 266..1208
/note="EF-1-alpha intron A"
/note="intron upstream of the start codon of human
EF-1-alpha"
CDS 1269..1988
/codon_start=1
/product="enhanced GFP"
/note="EGFP"
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
polyA_site 2140..2195
/note="beta-globin poly(A) signal"
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(2554..2570)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2578..2594
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(2602..2632)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 2647..2668
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2727..2922
/note="SV40 promoter"
/note="SV40 early promoter"
rep_origin 2773..2908
/note="SV40 ori"
/note="SV40 origin of replication"
polyA_signal 2928..3062
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(3301..3889)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4060..4920)
/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(4921..5025)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 gtcgacattg attattgact agatcatcgc gtgaggctcc ggtgcccgtc agtgggcaga
61 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc
121 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
181 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
241 caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct
301 ctttacgggt tatggccctt gcgtgccttg aattacttcc acgcccctgg ctgcagtacg
361 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt
421 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg
481 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt
541 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg
601 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg
661 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac
721 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg
781 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc
841 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg
901 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg
961 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta
1021 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg
1081 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt
1141 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc
1201 catttcaggt gtcgtgagga attctgcagt cgacggtacc gcgggcccgg gatccaccgg
1261 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
1321 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
1381 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
1441 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
1501 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
1561 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
1621 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
1681 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
1741 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
1801 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
1861 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
1921 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
1981 acaagtaaag cggccgcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg
2041 tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg
2101 gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca
2161 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
2221 atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat gccatatgct
2281 ggctgccatg aacaaaggtg gctataaaga ggtcatcagt atatgaaaca gccccctgct
2341 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
2401 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
2461 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atgaagatcc
2521 ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
2581 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
2641 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
2701 gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc
2761 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
2821 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
2881 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg
2941 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
3001 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
3061 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
3121 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
3181 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
3241 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
3301 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
3361 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
3421 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
3481 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
3541 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
3601 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
3661 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
3721 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
3781 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
3841 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
3901 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
3961 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
4021 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
4081 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
4141 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
4201 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
4261 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
4321 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
4381 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
4441 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
4501 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
4561 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
4621 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
4681 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
4741 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
4801 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
4861 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
4921 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
4981 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
5041 agtgccacct g
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