LOCUS V020239 7022 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020239 VERSION V020239 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 683..1702 /label="SLC51A(NM_152672)" /note="SLC51A(NM_152672)" /gene="SLC51A" CDS 1709..1774 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1798..2386 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2452..2500 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2702..3130 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3144..3473 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3540..4334 /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 4510..4631 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4680..4696) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4704..4720 /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(4728..4758) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4773..4794 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5082..5670) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5841..6701) /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(6702..6806) /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 gcgtttaaac ttaagcttgg 661 taccgagctc ggatccgcca ccatggagcc gggcaggacc cagataaagc ttgaccccag 721 gtacacagca gatcttctgg aggtgctgaa gaccaattac ggcatcccct ccgcctgctt 781 ctctcagcct cccacagcag cccaactcct gagagccctg ggccctgtgg aacttgccct 841 cactagcatc ctgaccttgc tggcgctggg ctccattgcc atcttcctgg aggatgccgt 901 ctacctgtac aagaacaccc tttgccccat caagaggcgg actctgctct ggaagagctc 961 ggcacccacg gtggtgtctg tgctgtgctg ctttggtctc tggatccctc gttccctggt 1021 gctggtggaa atgaccatca cctcgtttta tgccgtgtgc ttttacctgc tgatgctggt 1081 catggtggaa ggctttgggg ggaaggaggc agtgctgagg acgctgaggg acaccccgat 1141 gatggtccac acaggcccct gctgctgctg ctgcccctgc tgtccacggc tgctgctcac 1201 caggaagaag cttcagctgc tgatgttggg ccctttccaa tacgccttct tgaagataac 1261 gctgaccctg gtgggcctgt ttctcgtccc cgacggcatc tatgacccag cagacatttc 1321 tgaggggagc acagctctat ggatcaacac tttccttggc gtgtccacac tgctggctct 1381 ctggaccctg ggcatcattt cccgtcaagc caggctacac ctgggtgagc agaacatggg 1441 agccaaattt gctctgttcc aggttctcct catcctgact gccctacagc cctccatctt 1501 ctcagtcttg gccaacggtg ggcagattgc ttgttcgcct ccctattcct ctaaaaccag 1561 gtctcaagtg atgaattgcc acctcctcat actggagact tttctaatga ctgtgctgac 1621 acgaatgtac taccgaagga aagaccacaa ggttgggtat gaaactttct cttctccaga 1681 cctggacttg aacctcaaag ccctcgagga ctacaaagac catgacggtg attataaaga 1741 tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata tctcgacaat 1801 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct 1861 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg 1921 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg 1981 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt 2041 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt 2101 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg 2161 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc 2221 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat 2281 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc 2341 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga 2401 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa 2461 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc 2521 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt 2581 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac 2641 gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc 2701 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 2761 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 2821 acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt 2881 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg 2941 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 3001 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta 3061 taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt 3121 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc 3181 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt 3241 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 3301 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc 3361 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg 3421 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc 3481 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca 3541 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg 3601 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag 3661 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc 3721 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc 3781 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg 3841 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc 3901 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca 3961 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag 4021 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg 4081 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg 4141 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca 4201 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc 4261 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg 4321 acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac 4381 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc 4441 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc 4501 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 4561 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 4621 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg 4681 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 4741 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 4801 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 4861 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 4921 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 4981 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 5041 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 5101 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 5161 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 5221 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc 5281 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 5341 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 5401 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 5461 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 5521 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 5581 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 5641 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 5701 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 5761 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 5821 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 5881 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 5941 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 6001 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 6061 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 6121 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 6181 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 6241 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 6301 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 6361 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 6421 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 6481 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 6541 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 6601 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 6661 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 6721 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 6781 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac 6841 ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg 6901 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga 6961 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta 7021 gg //