LOCUS Exported 6095 bp ds-DNA circular SYN 08-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..6095
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 202..581
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 582..785
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 830..848
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 902..1618
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
polyA_signal 1662..1886
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1932..2360
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2374..2703
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2554..2689
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2770..3564
/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 3738..3859
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
protein_bind 3932..3948
/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(3956..3986)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4001..4022
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4310..4895)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5066..5926)
/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(5927..6031)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
901 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
961 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
1021 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
1081 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
1141 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
1201 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
1261 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
1321 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
1381 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
1441 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
1501 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
1561 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagct
1621 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
1681 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
1741 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
1801 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
1861 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
1921 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
1981 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
2041 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
2101 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
2161 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
2221 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
2281 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
2341 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
2401 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
2461 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
2521 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
2581 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
2641 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
2701 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
2761 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
2821 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
2881 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
2941 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
3001 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
3061 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
3121 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
3181 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
3241 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
3301 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
3361 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
3421 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
3481 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
3541 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
3601 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
3661 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
3721 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
3781 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
3841 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
3901 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
3961 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
4021 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
4081 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
4141 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
4201 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
4261 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
4321 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
4381 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
4441 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
4501 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
4561 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
4621 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
4681 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
4741 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
4801 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
4861 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
4921 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
4981 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
5041 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
5101 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
5161 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
5221 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
5281 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
5341 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
5401 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
5461 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
5521 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
5581 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
5641 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
5701 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
5761 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
5821 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
5881 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
5941 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
6001 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
6061 tcgacggatc gggagatctc ccgatcccct atggt
//