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pcDNA3.1-T2A-EGFP载体质粒基本信息
pcDNA3.1-T2A-EGFP质粒图谱载体图谱和pcDNA3.1-T2A-EGFP载体序列质粒序列多克隆位点信息
pcDNA3.1-T2A-EGFP质粒载体简介
pcDNA3.1-T2A-EGFP质粒序列载体序列
LOCUS pcDNA3.1-T2A-EGFP 6202 bp DNA circular SYN 13-JUL-2023
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6202)
AUTHORS Biofeng
TITLE Direct Submission
JOURNAL Exported Jul 13, 2023 from SnapGene 6.0.2
https://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6202
/mol_type="other DNA"
/organism="synthetic DNA construct"
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 991..1044
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 1045..1764
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
polyA_signal 1802..2026
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2072..2500
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2514..2843
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2694..2829
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2910..3704
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 3878..3999
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4048..4064)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 4072..4088
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(4096..4126)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4141..4162
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4450..5035)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5206..6066)
/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(6067..6171)
/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 aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc
961 agatatccag cacagtggcg gccgctcgag gagggcagag gaagtcttct aacatgcggt
1021 gacgtggagg agaatcccgg ccctatggtg agcaagggcg aggagctgtt caccggggtg
1081 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
1141 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
1201 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
1261 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
1321 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
1381 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
1441 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
1501 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
1561 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
1621 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
1681 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
1741 ggcatggacg agctgtacaa gtaatctaga gggcccgttt aaacccgctg atcagcctcg
1801 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
1861 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
1921 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
1981 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
2041 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
2101 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
2161 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
2221 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
2281 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
2341 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
2401 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
2461 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
2521 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
2581 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
2641 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
2701 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
2761 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
2821 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
2881 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
2941 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
3001 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
3061 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
3121 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
3181 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
3241 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
3301 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
3361 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
3421 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
3481 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
3541 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
3601 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
3661 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
3721 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
3781 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
3841 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
3901 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
3961 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
4021 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
4081 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
4141 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
4201 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
4261 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
4321 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
4381 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
4441 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
4501 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
4561 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
4621 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
4681 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
4741 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
4801 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
4861 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
4921 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
4981 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
5041 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
5101 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
5161 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
5221 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
5281 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
5341 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
5401 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
5461 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
5521 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
5581 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
5641 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
5701 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
5761 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
5821 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
5881 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
5941 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
6001 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
6061 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
6121 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
6181 ccgaaaagtg ccacctgacg tc
//
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