LOCUS V031265 5151 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031265 VERSION V031265 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..1298 /label="Gnrh1(NM_008145)" /note="Gnrh1(NM_008145)" /gene="Gnrh1" CDS 1317..1346 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 1365..1388 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" primer_bind complement(1408..1424) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" polyA_signal 1435..2057 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" rep_origin complement(2206..2794) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" polyA_signal 3123..3170 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" CDS complement(3402..4196) /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(4231..4588) /note="SV40 enhancer and early promoter" /label="SV40 promoter" promoter complement(4590..4694) /gene="bla" /label="AmpR promoter" rep_origin 4721..5149 /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 gatcctcaaa ctgatggccg gcattctact gctgactgtg tgtttggaag 1081 gctgctccag ccagcactgg tcctatgggt tgcgccctgg gggaaagaga aacactgaac 1141 acttggttga gtctttccaa gagatgggca aggaggtgga tcaaatggca gaaccccagc 1201 acttcgaatg tactgtccac tggccccgtt cacccctcag ggatctgcga ggagctctgg 1261 aaagtctgat tgaagaggaa gccaggcaga agaagatgac gcgtacgcgg ccgctcgagc 1321 agaaactcat ctcagaagag gatctggcag caaatgatat cctggattac aaggatgacg 1381 acgataaggt ttaaacggcc ggccgcggtc atagctgttt cctgaacaga tcccgggtgg 1441 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc 1501 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat 1561 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt 1621 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact 1681 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga 1741 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc 1801 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct 1861 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgatttta 1921 aaataactat accagcagga ggacgtccag acacagcata ggctacctgg ccatgcccaa 1981 ccggtgggac atttgagttg cttgcttggc actgtcctct catgcgttgg gtccactcag 2041 tagatgcctg ttgaattggg tacgcggcca gcggcgagcg gtatcagctc actcaaaggc 2101 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 2161 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 2221 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 2281 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 2341 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 2401 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 2461 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 2521 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 2581 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 2641 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 2701 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 2761 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 2821 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 2881 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 2941 atatgagtaa cctgaggcta tggcagggcc tgccgccccg acgttggctg cgagccctgg 3001 gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc 3061 ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt 3121 atgaacaaac gacccaacac cgtgcgtttt attctgtctt tttattgccg tcatagcgcg 3181 ggttccttcc ggtattgtct ccttccgtgt ttcagttagc ctccccctag ggtgggcgaa 3241 gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat 3301 tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt 3361 gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga 3421 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag 3481 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc 3541 tgatagcgat ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt 3601 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg 3661 ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg 3721 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga 3781 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt 3841 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc 3901 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca 3961 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt 4021 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac 4081 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat 4141 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga 4201 aacgatcctc atcctgtctc ttgatcgatc tttgcaaaag cctaggcctc caaaaaagcc 4261 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa 4321 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat 4381 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact 4441 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat 4501 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac 4561 attccacagc tggttctttc cgcctcagga ctcttccttt ttcaatatta ttgaagcatt 4621 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 4681 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca 4741 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 4801 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 4861 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 4921 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 4981 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 5041 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 5101 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt t //