LOCUS V032032 6056 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032032 VERSION V032032 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 941..1546 /label="PTTG1(NM_004219)" /note="PTTG1(NM_004219)" /gene="PTTG1" CDS 1553..1618 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1656..1880 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1926..2354 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2368..2697 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2764..3558 /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 3732..3853 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3902..3918) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3926..3942 /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(3950..3980) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3995..4016 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4304..4889) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5060..5920) /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(5921..6025) /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 atccgccacc atggctactc tgatctatgt 961 tgataaggaa aatggagaac caggcacccg tgtggttgct aaggatgggc tgaagctggg 1021 gtctggacct tcaatcaaag ccttagatgg gagatctcaa gtttcaacac cacgttttgg 1081 caaaacgttc gatgccccac cagccttacc taaagctact agaaaggctt tgggaactgt 1141 caacagagct acagaaaagt ctgtaaagac caagggaccc ctcaaacaaa aacagccaag 1201 cttttctgcc aaaaagatga ctgagaagac tgttaaagca aaaagctctg ttcctgcctc 1261 agatgatgcc tatccagaaa tagaaaaatt ctttcccttc aatcctctag actttgagag 1321 ttttgacctg cctgaagagc accagattgc gcacctcccc ttgagtggag tgcctctcat 1381 gatccttgac gaggagagag agcttgaaaa gctgtttcag ctgggccccc cttcacctgt 1441 gaagatgccc tctccaccat gggaatccaa tctgttgcag tctccttcaa gcattctgtc 1501 gaccctggat gttgaattgc cacctgtttg ctgtgacata gatattctcg aggactacaa 1561 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta 1621 gtgagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc 1681 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 1741 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 1801 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg 1861 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta 1921 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 1981 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 2041 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 2101 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 2161 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 2221 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 2281 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 2341 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 2401 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 2461 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 2521 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 2581 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 2641 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 2701 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt 2761 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 2821 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 2881 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 2941 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 3001 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 3061 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 3121 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 3181 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 3241 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 3301 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 3361 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 3421 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 3481 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 3541 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca 3601 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa 3661 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct 3721 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 3781 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 3841 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat 3901 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 3961 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 4021 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 4081 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 4141 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 4201 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 4261 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 4321 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 4381 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 4441 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 4501 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 4561 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 4621 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 4681 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 4741 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 4801 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc 4861 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 4921 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 4981 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 5041 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 5101 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 5161 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 5221 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 5281 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 5341 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 5401 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 5461 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 5521 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 5581 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 5641 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 5701 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 5761 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 5821 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 5881 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 5941 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 6001 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc //