LOCUS V030116 8395 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030116 VERSION V030116 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 202..581 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 582..785 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 830..848 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 874..1590 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK" /label="EGFP" CDS 1642..3885 /label="TAP1(NM_000593)" /note="TAP1(NM_000593)" /gene="TAP1" polyA_signal 3962..4186 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4232..4660 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4674..5003 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5070..5864 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 6038..6159 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6208..6224) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6232..6248 /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-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(6256..6286) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6301..6322 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6610..7195) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7366..8226) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(8227..8331) /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 ggctagcgcc accatggtga gcaagggcga ggagctgttc 901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc 961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc 1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg 1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg 1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc 1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc 1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac 1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc 1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc 1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc 1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg 1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc 1621 ggaggcggag ggagtgaatt catggctagc tctaggtgtc ccgctccccg cgggtgccgc 1681 tgcctccccg gagcttctct cgcatggctg gggacagtac tgctacttct cgccgactgg 1741 gtgctgctcc ggaccgcgct gccccgcata ttctccctgc tggtgcccac cgcgctgcca 1801 ctgctccggg tctgggcggt gggcctgagc cgctgggccg tgctctggct gggggcctgc 1861 ggggtcctca gggcaacggt tggctccaag agcgaaaacg caggtgccca gggctggctg 1921 gctgctttga agccattagc tgcggcactg ggcttggccc tgccgggact tgccttgttc 1981 cgagagctga tctcatgggg agcccccggg tccgcggata gcaccaggct actgcactgg 2041 ggaagtcacc ctaccgcctt cgttgtcagt tatgcagcgg cactgcccgc agcagccctg 2101 tggcacaaac tcgggagcct ctgggtgccc ggcggtcagg gcggctctgg aaaccctgtg 2161 cgtcggcttc taggctgcct gggctcggag acgcgccgcc tctcgctgtt cctggtcctg 2221 gtggtcctct cctctcttgg ggagatggcc attccattct ttacgggccg cctcactgac 2281 tggattctac aagatggctc agccgatacc ttcactcgaa acttaactct catgtccatt 2341 ctcaccatag ccagtgcagt gctggagttc gtgggtgacg ggatctataa caacaccatg 2401 ggccacgtgc acagccactt gcagggagag gtgtttgggg ctgtcctgcg ccaggagacg 2461 gagtttttcc aacagaacca gacaggtaac atcatgtctc gggtaacaga ggacacgtcc 2521 accctgagtg attctctgag tgagaatctg agcttatttc tgtggtacct ggtgcgaggc 2581 ctatgtctct tggggatcat gctctgggga tcagtgtccc tcaccatggt caccctgatc 2641 accctgcctc tgcttttcct tctgcccaag aaggtgggaa aatggtacca gttgctggaa 2701 gtgcaggtgc gggaatctct ggcaaagtcc agccaggtgg ccattgaggc tctgtcggcc 2761 atgcctacag ttcgaagctt tgccaacgag gagggcgaag cccagaagtt tagggaaaag 2821 ctgcaagaaa taaagacact caaccagaag gaggctgtgg cctatgcagt caactcctgg 2881 accactagta tttcaggtat gctgctgaaa gtgggaatcc tctacattgg tgggcagctg 2941 gtgaccagtg gggctgtaag cagtgggaac cttgtcacat ttgttctcta ccagatgcag 3001 ttcacccagg ctgtggaggt actgctctcc atctacccca gagtacagaa ggctgtgggc 3061 tcctcagaga aaatatttga gtacctggac cgcacccctc gctgcccacc cagtggtctg 3121 ttgactccct tacacttgga gggccttgtc cagttccaag atgtctcctt tgcctaccca 3181 aaccgcccag atgtcttagt gctacagggg ctgacattca ccctacgccc tggcgaggtg 3241 acggcgctgg tgggacccaa tgggtctggg aagagcacag tggctgccct gctgcagaat 3301 ctgtaccagc ccaccggggg acagctgctg ttggatggga agccccttcc ccaatatgag 3361 caccgctacc tgcacaggca ggtggctgca gtgggacaag agccacaggt atttggaaga 3421 agtcttcaag aaaatattgc ctatggcctg acccagaagc caactatgga ggaaatcaca 3481 gctgctgcag taaagtctgg ggcccatagt ttcatctctg gactccctca gggctatgac 3541 acagaggtag acgaggctgg gagccagctg tcagggggtc agcgacaggc agtggcgttg 3601 gcccgagcat tgatccggaa accgtgtgta cttatcctgg atgatgccac cagtgccctg 3661 gatgcaaaca gccagttaca ggtggagcag ctcctgtacg aaagccctga gcggtactcc 3721 cgctcagtgc ttctcatcac ccagcacctc agcctggtgg agcaggctga ccacatcctc 3781 tttctggaag gaggcgctat ccgggagggg ggaacccacc agcagctcat ggagaaaaag 3841 gggtgctact gggccatggt gcaggctcct gcagatgctc cagaatgagg atcctgatat 3901 ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg 3961 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc 4021 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt 4081 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat 4141 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa 4201 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg 4261 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 4321 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 4381 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 4441 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 4501 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 4561 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg 4621 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc 4681 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 4741 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 4801 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc 4861 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt 4921 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt 4981 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg 5041 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt 5101 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct 5161 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga 5221 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg 5281 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact 5341 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg 5401 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct 5461 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg 5521 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt 5581 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg 5641 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc 5701 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag 5761 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt 5821 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt 5881 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc 5941 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca 6001 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa 6061 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 6121 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac 6181 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 6241 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 6301 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 6361 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 6421 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 6481 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 6541 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 6601 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 6661 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 6721 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 6781 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 6841 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 6901 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 6961 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 7021 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa 7081 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 7141 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7201 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 7261 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 7321 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 7381 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 7441 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 7501 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 7561 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 7621 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 7681 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 7741 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 7801 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 7861 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 7921 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 7981 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8041 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 8101 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8161 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8221 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 8281 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 8341 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggt //