LOCUS V020069 7694 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020069 VERSION V020069 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..2374 /label="SLC1A6(NM_005071)" /note="SLC1A6(NM_005071)" /gene="SLC1A6" CDS 2381..2446 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2470..3058 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3124..3172 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3374..3802 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3816..4145 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4212..5006 /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 5182..5303 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5352..5368) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5376..5392 /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(5400..5430) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5445..5466 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5754..6342) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6513..7373) /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(7374..7478) /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 ccatgagcag ccatggcaac agcctgttcc tgcgggagag 721 cggccagcgg ctgggccggg tgggctggct gcagcggctg caggaaagcc tgcagcagag 781 agcactgcgc acgcgcctgc gcctgcagac catgaccctc gagcacgtgc tgcgcttcct 841 gcgccgaaac gccttcattc tgctgacggt cagcgccgtg gtcattgggg tcagcctggc 901 ctttgccctg cgcccatatc agctcaccta ccgccagatc aagtacttct cttttcctgg 961 agagcttctg atgaggatgc tgcagatgct ggtgttacct ctcattgtct ccagcctggt 1021 cacaggtatg gcatccctgg acaacaaggc cacggggcgg atggggatgc gggcagctgt 1081 gtactacatg gtgaccacca tcatcgcggt cttcatcggc atcctcatgg tcaccatcat 1141 ccatcccggg aagggctcca aggaggggct gcaccgggag ggccggatcg agaccatccc 1201 cacagctgat gccttcatgg acctgatcag aaatatgttt ccaccaaacc ttgtggaggc 1261 ctgcttcaaa cagttcaaga cgcagtacag cacgagggtg gtaaccagga ccatggtgag 1321 gacagagaac gggtctgagc cgggtgcctc catgcctcct ccattctcag tggagaacgg 1381 aaccagcttc ctggaaaatg tcactcgggc cttgggtacc ctgcaggaga tgctgagctt 1441 tgaggagact gtacccgtgc ctggctccgc caatggcatc aacgccctgg gcctcgtggt 1501 cttctctgtg gcctttgggc tggtcattgg tggcatgaaa cacaagggca gagtcctcag 1561 ggacttcttc gacagcctca atgaggctat tatgaggctg gtgggcatca ttatctggta 1621 tgcacctgtg ggcatcctgt tcctgattgc tgggaagatt ctggagatgg aagacatggc 1681 cgtcctgggg ggtcagctgg gcatgtacac cctgaccgtc atcgtgggcc tgttcctcca 1741 tgccggcatt gtccttcccc tcatctactt cctcgtcact caccggaacc ccttcccctt 1801 cattgggggc atgctacaag ccctcatcac cgctatgggc acgtcttcca gctcggcaac 1861 gctgcccatc accttccgct gcctggagga gggcctgggt gtggaccgcc gcatcaccag 1921 gttcgtcctg cccgtgggcg ccacggtcaa catggatggc actgccctct acgaggccct 1981 ggctgccatc ttcattgctc aagttaacaa ctacgagctc aacctgggtc agatcacaac 2041 catcagcatc acggccacag cagccagtgt tggggctgct ggcatccccc aggcgggtct 2101 ggtcaccatg gtcattgtgc ttacgtcggt cggcttgccc acggaagaca tcacgctcat 2161 cattgccgtg gactggttcc ttgaccggct tcgcacaatg accaacgtac tgggggactc 2221 aattggagcg gccgtcatcg agcacttgtc tcagcgggag ctggagcttc aggaagctga 2281 gcttaccctc cccagcctgg ggaaacccta caagtccctc atggcacagg agaagggggc 2341 atcccgggga cggggaggca acgagagtgc tatgctcgag gactacaaag accatgacgg 2401 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccga 2461 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 2521 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 2581 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 2641 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 2701 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 2761 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 2821 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca 2881 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 2941 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 3001 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 3061 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta 3121 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc 3181 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa 3241 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg 3301 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct 3361 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 3421 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 3481 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta 3541 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 3601 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 3661 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 3721 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt 3781 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt 3841 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 3901 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 3961 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 4021 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 4081 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 4141 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag 4201 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 4261 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 4321 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 4381 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 4441 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 4501 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 4561 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 4621 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 4681 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc 4741 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 4801 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 4861 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 4921 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 4981 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa 5041 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg 5101 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 5161 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc 5221 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 5281 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg 5341 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 5401 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 5461 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 5521 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 5581 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 5641 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 5701 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 5761 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 5821 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 5881 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 5941 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 6001 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 6061 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 6121 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 6181 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 6241 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 6301 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 6361 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 6421 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 6481 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 6541 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 6601 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 6661 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 6721 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 6781 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 6841 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 6901 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 6961 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 7021 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 7081 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 7141 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 7201 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 7261 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 7321 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 7381 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 7441 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 7501 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct 7561 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg 7621 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga 7681 agaatctgct tagg //