LOCUS V019889 6952 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019889 VERSION V019889 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..855 /label="COX8A(NM_004074)" /note="COX8A(NM_004074)" /gene="COX8A" CDS 883..1599 /codon_start=1 /note="mammalian codon-optimized" /product="enhanced YFP" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK" /label="EYFP" CDS 1639..1704 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1728..2316 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2382..2430 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2632..3060 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3074..3403 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3470..4264 /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 4440..4561 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4610..4626) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4634..4650 /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(4658..4688) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4703..4724 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5012..5600) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5771..6631) /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(6632..6736) /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 gtccgtcctg 661 acgccgctgc tgctgcgggg cttgacaggc tcggcccggc ggctcccagt gccgcgcgcc 721 aagatccatt cgttgccgcc ggaggggaag cttgggatca tggaattggc cgttgggctt 781 acctcctgct tcgtgacctt cctcctgcca gcgggctgga tcctgtcaca cctggagacc 841 tacaggaggc cagagggagg agggggaagc ggatccgcca ccatggtgag caagggcgag 901 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac 961 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag 1021 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc 1081 tacggcctga agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag 1141 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac 1201 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg 1261 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac 1321 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc 1381 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac 1441 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc 1501 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc 1561 gccgccggga tcactctcgg catggacgag ctgtacaagg aattctgcag atatccagca 1621 cagtggcggc cgctcgagga ctacaaagac catgacggtg attataaaga tcatgacatc 1681 gactacaagg atgacgatga caagtagtga gggcccgata tctcgacaat caacctctgg 1741 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat 1801 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt 1861 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca 1921 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg 1981 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg 2041 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca 2101 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca 2161 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc 2221 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc 2281 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga 2341 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa 2401 taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc 2461 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt 2521 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg 2581 caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct 2641 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg 2701 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg 2761 gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac 2821 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct 2881 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt 2941 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt 3001 tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt 3061 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag 3121 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc 3181 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc 3241 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 3301 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca 3361 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg 3421 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca 3481 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg 3541 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg 3601 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc 3661 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt 3721 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc 3781 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca 3841 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc 3901 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg 3961 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct 4021 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc 4081 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc 4141 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta 4201 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt 4261 ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac 4321 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 4381 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 4441 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 4501 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 4561 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 4621 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 4681 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 4741 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 4801 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 4861 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 4921 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 4981 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 5041 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 5101 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 5161 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta 5221 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 5281 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 5341 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 5401 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat 5461 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 5521 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 5581 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 5641 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 5701 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 5761 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 5821 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 5881 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 5941 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 6001 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 6061 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 6121 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 6181 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 6241 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 6301 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 6361 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 6421 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 6481 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 6541 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 6601 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 6661 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 6721 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag 6781 atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag 6841 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta 6901 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg //