LOCUS V031567 8397 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031567 VERSION V031567 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 927..3203 /label="Git2(NM_001347400)" /note="Git2(NM_001347400)" /gene="Git2" CDS 3240..3959 /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" polyA_signal 3997..4221 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4267..4695 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4709..5038 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5105..5899 /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 6073..6194 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6243..6259) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6267..6283 /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(6291..6321) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6336..6357 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6645..7230) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7401..8261) /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(8262..8366) /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 gtttaaagcc actgggatcc gccaccatgt cgaagcggct ccggagcagc gacgtgtgcg 961 ccgactgcaa cgggcctgat ccttcctggg cgtcagtaaa taggggaact ttcatatgtg 1021 acgaatgttg cagtgtccat cggagcctgg ggcgccatat ctcccaagtg aggcatctta 1081 aacacacagc atggcctcca accctgctcc agatggttga gaccttgtac aacaatggtg 1141 ctaactctat atgggaacac tctctgctgg accctgcctc tatcatgagt gggaggcgga 1201 aagctaaccc acaggacaaa gttcacccca ataaagcaga attcatcaga gccaagtatc 1261 agatgttagc gttcgtccac cgattgccct gccgcgagga cgacagcgtg acggccaaag 1321 atctcagtaa gcaactccat tcaagtgtga gaacagggaa tcttgagacc tgtctgagac 1381 tgttgtcttt aggagcccaa gccaacttct ttcatcctga aaaaggaagc actccactcc 1441 acgtcgcctc caaagcagga cagatcttac aggctgaatt gctggccgtg tatggagctg 1501 acccaggcac acaagattct agtgggaaga ctcctgttga ctatgcaagg caaggagggc 1561 accatgagtt ggcagaacgc ctcatagaaa tacagtacga gctgacggat agactggcct 1621 tctacctctg cggccggaaa ccagatcata aaagcggaca gcactttcta attcctcaga 1681 gggcagacag cagcctggat ttgtctgagt tggcaaaagc tgcgaagaag aaacttcaat 1741 ctctgagtaa tcatttgttt gaagaacttg ccatggatgt gtacgacgaa gttgacaggc 1801 gagagactga cgcagtctgg cttgccacgc agaaccacag taccttggta accgagacga 1861 ccgtcgtccc tttccttccg gtcaaccctg agtactcctc cacacgaaac cagggcagac 1921 agaaattagc tcggtttaac gcccacgagt ttgctacgct ggtcatcgac atcctcagtg 1981 atgccaagag gagacaacag ggcagtcctc tgtccaggtc aaaagacaat gtggagctca 2041 tattgagaac agtcagtacc cagcacagca ctgagagcca agacaacgac cagccagact 2101 atgacagtgt ggcctccgat gaagacacag acgtggagac gagggcaagc aaggcaaacc 2161 ggcagaagag cctagattca gatttgtcag atggaccagt cactgtgcag gaatttatgg 2221 aggtcaaaag cgccctagtg gcttctgagg ccaagagaca gcagctaatg aaggtgaata 2281 acaacttgag tgacgagctg agaattatgc agaaaaagct tcaaactctc cagagtgaga 2341 actcaagcct caggagacag gccacagcca gtgcctgtca ggtgcagact ggctctgacc 2401 acaaggacac tgcaagccac tcgtccctaa agcgacggcc atctgcccgg ggcagtagac 2461 ccatgtccat gtacgagact ggatcaggcc agaagccata cctcccaatg ggagaagcca 2521 gccaccctga ggagagccgg acgagacttc aacccttccc cacgcatatt gggaggagtg 2581 cacttgtgac ctcctcttcg tctctgccct ccttcccctc cacactttcc tggtccaggg 2641 acgaaagcgc ccgaagggcc tccagactgg agaagcagaa cagcaccccg gagagtgatt 2701 atgacaacac tgcctgtgac cctgagccag acgacacggg gtcaacccgg aagggacggc 2761 agcggagcat gctctggcaa ggtgatggct tattgccaga tacagcagaa ccccactcgg 2821 tccccagccc caccctcccc agcacagaag acgtcatccg gaagaccgag cagatcacca 2881 aaaacatcca ggagctcctg cgagcagccc aggagaacaa gcatgacagc tatattccct 2941 gctcagagag gatacacgtt gctgtaaccg agatggcagc cctgttcccc aaaaaaccca 3001 agtctgacac ggtgaggact tccctccgcc tactcacgtc cagtgcctac cggctccagt 3061 cggagtgcag aaaggctctc ccaggagact ccagcttacc cacagatgtg cagctggtca 3121 cacagcaggt catccagtgt gcctatgaca tcgccaaggc cgccaaacag ctggtcacaa 3181 tcaccaccaa agagaacagc agctcgagtg gcggtggagg aagcggcgga ggtggatcta 3241 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg 3301 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg 3361 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc 3421 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc 3481 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct 3541 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg 3601 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca 3661 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg 3721 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg 3781 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact 3841 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc 3901 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtagt 3961 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat 4021 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc 4081 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg 4141 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg 4201 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt 4261 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg 4321 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc 4381 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc 4441 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta 4501 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta 4561 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg 4621 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa 4681 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg 4741 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg 4801 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 4861 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca 4921 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc 4981 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa 5041 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt 5101 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct 5161 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct 5221 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga 5281 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc 5341 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg 5401 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc 5461 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca 5521 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga 5581 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc 5641 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga 5701 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca 5761 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg 5821 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct 5881 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc 5941 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga 6001 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc 6061 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 6121 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 6181 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca 6241 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 6301 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 6361 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 6421 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 6481 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 6541 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 6601 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 6661 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 6721 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 6781 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 6841 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 6901 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 6961 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 7021 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 7081 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 7141 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag 7201 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 7261 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 7321 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 7381 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 7441 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 7501 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 7561 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 7621 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 7681 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 7741 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 7801 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 7861 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 7921 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 7981 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 8041 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 8101 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 8161 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 8221 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 8281 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 8341 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc //