LOCUS Exported 6172 bp ds-DNA circular SYN 28-SEP-2017
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pECMV-3×FLAG-VENUS
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6172)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 28, 2017 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6172
/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"
CDS 1009..1728
/codon_start=1
/label=VENUS
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
polyA_signal 1772..1996
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2042..2470
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2484..2813
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2664..2799
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2880..3674
/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 3848..3969
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(4420..5005)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5176..6036)
/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(6037..6141)
/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 ggtgtccaag
1021 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
1081 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
1141 ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
1201 ctgggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
1261 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
1321 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
1381 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
1441 aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc
1501 aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag
1561 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
1621 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
1681 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtgact cgagtctaga
1741 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
1801 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
1861 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
1921 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
1981 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
2041 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
2101 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
2161 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
2221 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
2281 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
2341 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
2401 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
2461 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
2521 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
2581 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
2641 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
2701 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
2761 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
2821 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
2881 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
2941 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
3001 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
3061 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
3121 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
3181 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
3241 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
3301 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
3361 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
3421 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
3481 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
3541 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
3601 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
3661 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
3721 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
3781 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
3841 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
3901 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
3961 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
4021 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
4081 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
4141 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
4201 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
4261 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
4321 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
4381 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
4441 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
4501 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
4561 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
4621 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
4681 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
4741 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
4801 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
4861 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
4921 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
4981 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
5041 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
5101 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
5161 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
5221 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
5281 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
5341 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
5401 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
5461 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
5521 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
5581 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
5641 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
5701 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
5761 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
5821 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
5881 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
5941 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
6001 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
6061 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
6121 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//