LOCUS V022676 8581 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022676 VERSION V022676 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 649..1365 /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 1387..3324 /label="Cep72(NM_028959)" /note="Cep72(NM_028959)" /gene="Cep72" misc_feature 3357..3945 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 4011..4059 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 4261..4689 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4703..5032 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5099..5893 /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 6069..6190 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6239..6255) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6263..6279 /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(6287..6317) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6332..6353 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6641..7229) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7400..8260) /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(8261..8365) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgccaccat ggtgagcaag 661 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac 721 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc 781 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc 841 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc 901 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac 961 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc 1021 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac 1081 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg 1141 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag 1201 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc 1261 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc 1321 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggagg agggggaagc 1381 ggtaccatgg cgccaggcca gcggttggtt ttgtgtgaag agactgtacg agagcgcagc 1441 ggcctgggtc cccaccgcga cctggctgaa cttcgatctt tatctattcc tggaacatac 1501 caagagaaga ttactcacct gggaaactct ctgatgcact tgacagctct aaaatcctta 1561 gacctctcac gtaactcctt ggttagcctt gagggtatac agtatctggt ctccttggag 1621 agtctcaatc tctactacaa ctgtatttct tcattagcag aagtatttag gctccacacc 1681 ctactggagc tccaggacgt ggacttccgg cttaaccccg ttgtgaagaa tgagtctgac 1741 taccgcctgt ttgtcgtgca catgctcccg aagctccgac agctagatga ccgtcctgtg 1801 agagaaagtg agcggaaagc ttcccagctt cattttgccc cagaagattc actgaactca 1861 aaggagaact tctcaaccac tttaaccgtt ggaagaccac accacctcag aaacagatgt 1921 actgagacct cggccaaaaa atgcttggtc atggatgctg atgatgaggc tgtcctgaac 1981 cttattgccg agtgtgaatg ggacctaagc aatcctcctg ggaacatgag ctccagccaa 2041 aaggaacatg aagctgacct ccattatgcc caagagtcca gacatctgtt gagtcctttg 2101 tccatccagc accagtgtgg ggattcagcg aggcgaggcc atgagaagaa gaaagtcacc 2161 tccagagggt gtccaggtca ttcaccgcag gaccagctct gtggggaact ccccctacag 2221 catggcttgc cagaggcctg ccatatgcac gtacagcatg cccgcatcac ttcacagcca 2281 gactctgtgg atgttgagga ctgtgcttca tctgctcaga aatcaagttt gtcatcacaa 2341 aaaacagtaa atcctttgcc tgtccctgaa aagtacagga agcgaaggat gcctggtggg 2401 agattccagg tgccttcaga ccgggagtgt ttgagcttcc tagagagggc tgacgggccc 2461 tccagccttg aggacagtct aagcagacag gatggcttgg agggccagag tcaggtggcc 2521 ttgtctcact ctgaggcttt ggaggctgaa gagcggacat ctcatggcac aagtgacccc 2581 agagtgctgt ctcctaaact ctgctcggca gcggtgcctg agcggaggtc cactctggaa 2641 gtggcccttc tggaggcgct tctcgacctg atagacaggt gctcaagtgg cagtggatcc 2701 ctgcatggca atgaggcctt ccttgctcag gccaagcatg ttttgtcatc tcttcaagag 2761 ttcacagcaa ctcgggacag ttcagcactt gaaaaggaag gaattggtta cctatctctt 2821 gaaaataaga ctctgcaaag ccgccttgct gagcaacagc aacagtatac tgctacagtg 2881 accaagatga cagcagagct caataataca aagagggagc tggacacatt gagacaacat 2941 ttggacaaat cattggaaga gaatagccac ttaaagtctc ttctgtacaa tgtgaaaaaa 3001 gaagtaaaga ctgcagacac ttcaactgca ctcactttgc agatcactgg acttcaggca 3061 agtatgaagc agctctctgg ggaagttgtg gagctgaagc aacacgtgga gcactatgac 3121 aagatccagg agctcacaca gatgcttcaa gagagccaca gctccctggt aagcaccaat 3181 gagcatctcc tgcaggagct gagccgcaca cgggcacagc atagagctga ggtagagcag 3241 atgcgctgga gcttccagga atttaagaag actacggccc tgatccctca ccgtagctcc 3301 cgacgtgggg gccggcagtc ctgctagctc gagtctagag ggcccgatat ctcgacaatc 3361 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt 3421 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg 3481 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc 3541 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt 3601 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg 3661 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg 3721 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct 3781 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc 3841 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc 3901 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag 3961 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa 4021 aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc 4081 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt 4141 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg 4201 tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc 4261 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 4321 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 4381 cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta 4441 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc 4501 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 4561 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat 4621 aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta 4681 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc 4741 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc 4801 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat 4861 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc 4921 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga 4981 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc 5041 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat 5101 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg 5161 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc 5221 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca 5281 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct 5341 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga 5401 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg 5461 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat 5521 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga 5581 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg 5641 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg 5701 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat 5761 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct 5821 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga 5881 cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc 5941 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg 6001 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg 6061 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 6121 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 6181 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt 6241 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 6301 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 6361 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 6421 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 6481 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 6541 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 6601 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 6661 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 6721 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 6781 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct 6841 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 6901 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 6961 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 7021 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 7081 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 7141 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 7201 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 7261 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 7321 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 7381 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 7441 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 7501 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 7561 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 7621 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 7681 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 7741 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 7801 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 7861 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 7921 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 7981 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 8041 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 8101 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 8161 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 8221 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 8281 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 8341 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg 8401 gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc 8461 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag 8521 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag 8581 g //