LOCUS V031027 8271 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031027 VERSION V031027 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers primer_bind 166..182 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" enhancer 343..722 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 723..926 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 952..970 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 1029..4418 /label="Srebf2(NM_033218)" /note="Srebf2(NM_033218)" /gene="Srebf2" CDS 4437..4466 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 4485..4508 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" primer_bind complement(4528..4544) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" polyA_signal 4555..5177 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" rep_origin complement(5326..5914) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" polyA_signal 6243..6290 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" CDS complement(6522..7316) /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" promoter complement(7351..7708) /note="SV40 enhancer and early promoter" /label="SV40 promoter" promoter complement(7710..7814) /gene="bla" /label="AmpR promoter" rep_origin 7841..8269 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" ORIGIN 1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag 61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa 121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca 181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt 241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata 301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta 361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat 781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac 901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac 961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg 1021 cgatcgccat ggatgagagc agcgagctgg gcgttctgga gaccatggag accctcacgg 1081 agctgggcga tgagctgact ctcggggaca tcgacgagat gctacagttt gtcagcaatc 1141 aagtgggaga gttccctgat ttgttttcag agcaactttg tagctccttc cctggaggtg 1201 gcagcaatgg tggcagtggc aacaacagca gtggcagagg caacaatggt ggagccacag 1261 accctgcagt acagcggtca ttcagccagg tcccattatc taccttctct ccctcagctg 1321 cctcccctca ggctccggcc ctgcaggtca aagtctctcc taccccacca agggcaaccc 1381 ctgttctcca gccgcgaccc cagccccagc cccaacctcc agctcagctg cagcagcaga 1441 cggtgatgat caccccgacg ttcagcaccg ctccgcagac gaggatcatc cagcagcctt 1501 tgatatacca gaacgcagcg accagctttc aagtcctgca gcctcaagtg caaagcctcg 1561 tgacgtcccc gcaggtgcag cccgtcacca tccagcagca ggtgcagacg gtacaggccc 1621 agcgggtgtt gacgcagaca gccaatggca cactgcagac ccttgccccc gccacggtac 1681 agacggtcgc tgcgccacag gtgcagcagg tcccggtcct ggtgcaacct cagatcatta 1741 agacagattc ccttgtgttg accacgctga agacagatgg cagccctgtc atggctgcag 1801 tccagaaccc ggccctcacg gccctcactg cccccatcca gacagccgcc cttcaagtgc 1861 caaccctggt gggcagcaac gggaccattc tgaccacaat gcctgtgatg atggggcaag 1921 agaaagttcc tatcaagcaa gtgcctggcg gcgtgaagca gctggatcct cccaaagaag 1981 gagagaggcg gacaacacac aatatcattg aaaagcgcta ccggtcctcc atcaacgaca 2041 aaatcataga gttgaaggac ttagtcatgg ggacagatgc caagatgcac aagtctggcg 2101 ttctgaggaa ggccattgat tacatcaaat atctgcagca ggtcaatcac aagctgcgcc 2161 aggagaacat ggtgctgaag ctggccaatc agaaaaacaa gctcctgaag ggcatcgacc 2221 tgggcagtct ggtggacagt gatgtggact tgaaaattga tgactttaac cagaatgtcc 2281 ttctgatgtc tccgccggcc tccgactccg ggtcccaggc cggcttctct ccctattcca 2341 ttgactctga gccgggcagc cctctgctgg atgacgcaaa ggtcaaggat gaaccggact 2401 ctcctcctgt ggcactgggc atggtggacc gctctcgaat cctcttatgt gtcctcacct 2461 tcctgggcct ctcctttaac cccttgactt ccttgctgca gtggggaggg gcccacaaca 2521 ctgaccagca cccatactca ggctcgggcc gcagtgtact gtcactggag tcaggtgctg 2581 ggggctggtt tgactggatg gttccaactc tcctcctgtg gctggtaaat ggtgtgattg 2641 tcttgagcgt ctttgtgaag ctgttggtcc acggggagcc ggtgattcgc ccacactcac 2701 gcccctcagt caccttctgg agacaccgga agcaggcaga cctagacctc gccaaaggtg 2761 atttcgcagc tgctgctgcc aacctacaaa cctgcttatc cgtgctgggc cgggcgctgc 2821 ccacctcccg cctggacttg gcctgcagcc tctcctggaa tgtgatccgc tacagcctgc 2881 agaagctgcg cctggtacgc tggttactca agaaggtctt ccagcgctgg cgggctactc 2941 ccgccactgc agctggcttt gaggatgagg ctaagagcag cgcaagggat gcggcactgg 3001 cctaccacag gctgcaccag ctgcacatca cagggaagct tccggcggga tctgcctgtt 3061 cagatgtgca catggctctg tgtgctgtga acctggccga gtgtgcggag gagaaaatcc 3121 taccaagcac actgattgag atccacctga cagcggccat ggggctcaag acccgctgtg 3181 gaggcaagct gggcttcttg gctagctact tcttaaatcg tgcccagagt ctgtgtggcc 3241 cagagcatag cactgtcccc gactccctgc gctggctatg ccatcccttg ggccagaagt 3301 ttttcatgga gcgaagctgg tcgatcaagt cagcagccaa ggagagcctg tactgcgccc 3361 agaggagccc agctgacccc attgcccagg tacaccaggc gttctgcaaa aacctgctag 3421 aaagagcggt ggagtccttg gtgaaacctc aggccaagaa gaaggcaggc gaccaggaag 3481 aagagagctg tgaattttcc agtgctctag agtatttgaa attacttcat tcttttgtgg 3541 actctgtggg atttgtgacc tcaccattct ccagcagttc cgtgctcagg tcggcccttg 3601 gccccgatgt catctgccgg tggtggacgt ccgcagtcac catggccatc agctggctcc 3661 agggagacga tgcagctgtg cgctctcgtt ttactgaagt agagcgggtc cccaaggccc 3721 tggaagtgac cgagagtccc ttggtgaagg ccgtcttcta tacctgtaga gccatgcatg 3781 cctccctctc tgggaaagct gacgggcagc agaactcctt ctgtcactgt gagagggcca 3841 gtggtcacct gtggagcagt ctcaacgtca gcggcactac ttctgatcct tcacttaacc 3901 atgtgatcca gctgttcacc tgtgacctgc tactgtcgct gaggacagca ctctggcaaa 3961 agcaggccag tgccagccag ctcctgggtg agacctacca tgcatccggc actgagctgg 4021 ccggcttcca gcgggacctg ggcagccttc gaaggctggc ccatagcttc cgcccagcct 4081 accgcaaggt gttcctgcat gaagccactg tgcgcctgat ggcaggagcc agccctaccc 4141 gtacacacca gctgctggag catagcctac ggcggcgccc cactcagaac accaagcatg 4201 gagaggtaga cacctggcct ggccaacgag agcgagctac tgccatcctg ctggcctgcc 4261 gccatcttcc cctctctttc ctgtcttccc cgggccagcg ggcagtgctg ctggctgaag 4321 ctgcccgcac cctggagaag gtgggtgacc gccgttcctg cagtgactgc cagcagatga 4381 ttgtcaagct gggcggtggc actgccatcg ctgcctccac gcgtacgcgg ccgctcgagc 4441 agaaactcat ctcagaagag gatctggcag caaatgatat cctggattac aaggatgacg 4501 acgataaggt ttaaacggcc ggccgcggtc atagctgttt cctgaacaga tcccgggtgg 4561 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc 4621 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat 4681 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt 4741 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact 4801 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga 4861 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc 4921 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct 4981 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgatttta 5041 aaataactat accagcagga ggacgtccag acacagcata ggctacctgg ccatgcccaa 5101 ccggtgggac atttgagttg cttgcttggc actgtcctct catgcgttgg gtccactcag 5161 tagatgcctg ttgaattggg tacgcggcca gcggcgagcg gtatcagctc actcaaaggc 5221 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 5281 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 5341 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 5401 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 5461 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 5521 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 5581 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 5641 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 5701 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 5761 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 5821 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 5881 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 5941 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 6001 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 6061 atatgagtaa cctgaggcta tggcagggcc tgccgccccg acgttggctg cgagccctgg 6121 gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc 6181 ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt 6241 atgaacaaac gacccaacac cgtgcgtttt attctgtctt tttattgccg tcatagcgcg 6301 ggttccttcc ggtattgtct ccttccgtgt ttcagttagc ctccccctag ggtgggcgaa 6361 gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat 6421 tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt 6481 gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga 6541 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag 6601 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc 6661 tgatagcgat ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt 6721 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg 6781 ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg 6841 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga 6901 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt 6961 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc 7021 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca 7081 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt 7141 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac 7201 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat 7261 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga 7321 aacgatcctc atcctgtctc ttgatcgatc tttgcaaaag cctaggcctc caaaaaagcc 7381 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa 7441 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat 7501 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact 7561 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat 7621 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac 7681 attccacagc tggttctttc cgcctcagga ctcttccttt ttcaatatta ttgaagcatt 7741 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 7801 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca 7861 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 7921 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 7981 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 8041 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 8101 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 8161 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 8221 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt t //