LOCUS V038240 6866 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038240 VERSION V038240 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 923..1639 /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 1676..2356 /label="NAT8(NM_003960)" /note="NAT8(NM_003960)" /gene="NAT8" CDS 2363..2428 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2466..2690 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2736..3164 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3178..3507 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3574..4368 /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 4542..4663 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4712..4728) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4736..4752 /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(4760..4790) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4805..4826 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5114..5699) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5870..6730) /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(6731..6835) /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 aagcttgcca ccatggtgag caagggcgag gagctgttca ccggggtggt 961 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga 1021 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa 1081 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag 1141 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 1201 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt 1261 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 1321 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat 1381 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga 1441 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 1501 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa 1561 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg 1621 catggacgag ctgtacaagg gcggtggagg cagtggtgga ggcggtagcg aattcatggc 1681 tccttgtcac atccgcaaat accaggagag cgaccgccag tgggttgtgg gcttgctctc 1741 ccgggggatg gccgagcatg ccccagccac cttccggcaa ttgctgaagc tgcctcgaac 1801 cctcatactc ttacttgggg ggcccctcgc cctactcctg gtctctggat cctggcttct 1861 agccctcgtg ttcagcatca gcctcttccc tgccctgtgg ttccttgcca aaaaaccctg 1921 gacggagtat gtggacatga cattgtgcac agacatgtct gacattacca aatcctacct 1981 gagtgagcgt ggctcctgct tctgggtggc tgagtctgaa gagaaggtgg tgggcatggt 2041 aggagctctg cctgttgatg atcccacctt gagggagaag cggttgcagc tgtttcatct 2101 ctttgtggac agtgagcacc gtcgtcaggg gatagcaaaa gccctggtca ggactgtcct 2161 ccagtttgcc cgggaccagg gctacagtga agttatcctg gacaccggca ccatccagct 2221 ctctgctatg gccctctacc agagcatggg cttcaagaag acgggccagt ccttcttctg 2281 tgtgtgggcc aggctagtgg ctcttcatac agttcatttc atctaccacc tcccttcttc 2341 taaggtaggg agtctgctcg aggactacaa agaccatgac ggtgattata aagatcatga 2401 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc 2461 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 2521 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 2581 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 2641 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 2701 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag 2761 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 2821 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 2881 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 2941 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 3001 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 3061 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta 3121 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg 3181 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 3241 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 3301 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 3361 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 3421 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc 3481 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga 3541 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca 3601 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 3661 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 3721 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg 3781 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 3841 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 3901 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 3961 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 4021 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 4081 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc 4141 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 4201 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 4261 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 4321 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg 4381 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 4441 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 4501 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt 4561 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 4621 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 4681 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 4741 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 4801 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 4861 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 4921 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 4981 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 5041 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 5101 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 5161 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 5221 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 5281 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 5341 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 5401 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 5461 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 5521 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 5581 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 5641 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 5701 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 5761 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 5821 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 5881 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 5941 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 6001 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 6061 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 6121 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 6181 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 6241 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 6301 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 6361 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 6421 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 6481 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 6541 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 6601 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 6661 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 6721 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 6781 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 6841 ttccccgaaa agtgccacct gacgtc //