LOCUS V032432 6061 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032432 VERSION V032432 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 964..1551 /label="GINS1(NM_021067)" /note="GINS1(NM_021067)" /gene="GINS1" CDS 1558..1623 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1661..1885 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1931..2359 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2373..2702 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2769..3563 /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 3737..3858 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3907..3923) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3931..3947 /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(3955..3985) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4000..4021 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4309..4894) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5065..5925) /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(5926..6030) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc 961 accatgttct gcgaaaaagc catggaactg atccgcgagc tgcatcgcgc gcccgaaggg 1021 caactgcctg ccttcaacga ggatggactc agacaagttc tggaggagat gaaagctttg 1081 tatgaacaaa accagtctga tgtgaatgaa gcaaagtcag gtggacgaag tgatttgata 1141 ccaactatca aatttcgaca ctgttctctg ttaagaaatc gacgctgcac tgtagcatac 1201 ctgtatgacc gcttgcttcg gatcagagca ctcagatggg aatatggtag cgtcttgcca 1261 aatgcattac gatttcacat ggctgctgaa gaaatggagt ggtttaataa ttataaaaga 1321 tctcttgcta cttatatgag gtcactggga ggagatgaag gtttggacat tacacaggat 1381 atgaaaccac caaaaagcct atatattgaa gtccggtgtc taaaagacta tggagaattt 1441 gaagttgatg atggcacttc agtcctatta aaaaaaaata gccagcactt tttacctcga 1501 tggaaatgtg agcagctgat cagacaagga gtcctggagc acatcctgtc actcgaggac 1561 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac 1621 aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag 1681 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact 1741 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt 1801 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat 1861 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg 1921 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 1981 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 2041 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg 2101 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 2161 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 2221 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct 2281 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa 2341 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc 2401 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt 2461 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 2521 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 2581 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 2641 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 2701 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat 2761 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga 2821 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc 2881 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga 2941 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg 3001 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc 3061 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg 3121 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga 3181 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc 3241 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca 3301 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg 3361 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct 3421 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg 3481 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc 3541 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac 3601 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt 3661 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga 3721 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag 3781 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 3841 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta 3901 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 3961 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 4021 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 4081 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 4141 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 4201 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 4261 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 4321 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 4381 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 4441 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 4501 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 4561 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 4621 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 4681 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 4741 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 4801 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa 4861 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 4921 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 4981 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 5041 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 5101 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 5161 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 5221 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 5281 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 5341 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 5401 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 5461 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 5521 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 5581 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 5641 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 5701 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 5761 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 5821 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 5881 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 5941 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 6001 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 6061 c //