LOCUS V047481 7550 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V047481 VERSION V047481 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 890..2299 /label="MMP12(NM_002426)" /note="MMP12(NM_002426)" /gene="MMP12" CDS 2336..3052 /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 3059..3076 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" polyA_signal 3117..3341 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3387..3815 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3829..4158 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4225..5019 /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 5193..5314 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5363..5379) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5387..5403 /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(5411..5441) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5456..5477 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5765..6350) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6521..7381) /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(7382..7486) /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 cttgccacca tgaagtttct 901 tctaatactg ctcctgcagg ccactgcttc tggagctctt cccctgaaca gctctacaag 961 cctggaaaaa aataatgtgc tatttggtga aagatactta gaaaaatttt atggccttga 1021 gataaacaaa cttccagtga caaaaatgaa atatagtgga aacttaatga aggaaaaaat 1081 ccaagaaatg cagcacttct tgggtctgaa agtgaccggg caactggaca catctaccct 1141 ggagatgatg cacgcacctc gatgtggagt ccccgatgtc catcatttca gggaaatgcc 1201 aggggggccc gtatggagga aacattatat cacctacaga atcaataatt acacacctga 1261 catgaaccgt gaggatgttg actacgcaat ccggaaagct ttccaagtat ggagtaatgt 1321 tacccccttg aaattcagca agattaacac aggcatggct gacattttgg tggtttttgc 1381 ccgtggagct catggagact tccatgcttt tgatggcaaa ggtggaatcc tagcccatgc 1441 ttttggacct ggatctggca ttggagggga tgcacatttc gatgaggacg aattctggac 1501 tacacattca ggaggcacaa acttgttcct cactgctgtt cacgagattg gccattcctt 1561 aggtcttggc cattctagtg atccaaaggc cgtaatgttc cccacctaca aatatgttga 1621 catcaacaca tttcgcctct ctgctgatga catacgtggc attcagtccc tgtatggaga 1681 cccaaaagag aaccaacgct tgccaaatcc tgacaattca gaaccagctc tctgtgaccc 1741 caatttgagt tttgatgctg tcactaccgt gggaaataag atctttttct tcaaagacag 1801 gttcttctgg ctgaaggttt ctgagagacc aaagaccagt gttaatttaa tttcttcctt 1861 atggccaacc ttgccatctg gcattgaagc tgcttatgaa attgaagcca gaaatcaagt 1921 ttttcttttt aaagatgaca aatactggtt aattagcaat ttaagaccag agccaaatta 1981 tcccaagagc atacattctt ttggttttcc taactttgtg aaaaaaattg atgcagctgt 2041 ttttaaccca cgtttttata ggacctactt ctttgtagat aaccagtatt ggaggtatga 2101 tgaaaggaga cagatgatgg accctggtta tcccaaactg attaccaaga acttccaagg 2161 aatcgggcct aaaattgatg cagtcttcta ctctaaaaac aaatactact atttcttcca 2221 aggatctaac caatttgaat atgacttcct actccaacgt atcaccaaaa cactgaaaag 2281 caatagctgg tttggttgtg gtggaggcgg atcaggaggc ggtggatctg gatccatggt 2341 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga 2401 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa 2461 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt 2521 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca 2581 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa 2641 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa 2701 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct 2761 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat 2821 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca 2881 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct 2941 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct 3001 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agctcgagca 3061 ccatcaccat caccattagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt 3121 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 3181 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 3241 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 3301 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac 3361 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg 3421 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 3481 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 3541 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 3601 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 3661 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 3721 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa 3781 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta 3841 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 3901 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 3961 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 4021 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 4081 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga 4141 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa 4201 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 4261 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 4321 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 4381 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg 4441 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 4501 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 4561 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 4621 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 4681 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 4741 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 4801 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 4861 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 4921 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 4981 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa 5041 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 5101 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 5161 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt 5221 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 5281 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta 5341 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 5401 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 5461 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 5521 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 5581 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 5641 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 5701 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 5761 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 5821 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 5881 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 5941 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 6001 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 6061 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 6121 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 6181 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 6241 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 6301 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 6361 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 6421 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 6481 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 6541 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 6601 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 6661 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 6721 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 6781 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 6841 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 6901 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 6961 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 7021 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 7081 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 7141 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 7201 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 7261 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 7321 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 7381 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 7441 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 7501 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt //