LOCUS V023842 7736 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023842 VERSION V023842 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..3226 /label="CEP85(NM_022778)" /note="CEP85(NM_022778)" /gene="CEP85" CDS 3233..3298 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3336..3560 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3606..4034 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4048..4377 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4444..5238 /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 5412..5533 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5582..5598) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5606..5622 /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(5630..5660) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5675..5696 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5984..6569) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6740..7600) /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(7601..7705) /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 atggccatgc aggagaaata 961 tccaactgag gggatctctc acgtcacttc accgagttcc gatgtgattc agaagggcag 1021 ttccctgggg actgaatggc agaccccagt tatctcggag ccctttcgga gccgcttcag 1081 ccgctgttca agtgtagccg acagtgggga cacagccatt ggtacatcat gctcagatat 1141 tgcggaggat ttttgcagct caagtggcag tcctcctttc cagcccatca aaagccacgt 1201 aaccattcca acagcccatg tgatgccttc tactttaggg acctctcctg ccaagccaaa 1261 ttctacacct gttggaccct cttcctctaa actccctttg tcagggttgg ctgaaagtgt 1321 gggaatgaca agaaatggag acctcggtgc aatgaaacat tctccaggcc tatctagaga 1381 tctcatgtat ttctctggtg ctactggaga aaatggaatt gagcagtcct ggtttccagc 1441 agtgggccat gaaagacaag aagaggcgag gaagtttgat attcctagca tggaatctac 1501 cctcaatcag tcggcaatga tggagacact ttattcagat cctcaccacc gagtccgctt 1561 ccacaaccca agaaccagca caagtaagga gttgtacaga gtgttgcctg aggccaagaa 1621 ggcaccgggc agcggggcag tgtttgagcg gaatggacca cattctaata gcagtggggt 1681 cctccctttg ggactccagc ctgctcccgg gctctccaag cctctgccct ctcaggtgtg 1741 gcagccgagt cctgacactt ggcatccccg agagcaatct tgtgaactca gcacttgtcg 1801 gcagcagctg gaattgattc gtttacagat ggagcaaatg cagcttcaga atggagccat 1861 ctgccaccat cctgctgctt ttggtccttc actgcccatc ttagagccag cacagtggat 1921 cagcatcttg aacagtaatg aacaccttct gaaggaaaaa gagcttctca ttgacaagca 1981 gaggaaacac atctctcagc tggagcagaa agtgcgagag agcgaactgc aagtccacag 2041 tgccctcttg ggccgccctg ccccctttgg tgatgtctgc ttgctgaggc tacaggaatt 2101 gcagcgagaa aacactttct tacgtgcaca gtttgcacag aagacagaag ccttgagcag 2161 agaaaagatt gaccttgaaa agaaactctc tgcttctgaa gttgaagtcc agctcatcag 2221 agagtcgctc aaagtggcgt tgcagaagca ttctgaggaa gtgaagaaac aggaagaaag 2281 ggtcaaaggt cgtgataaac atatcaataa tttgaaaaag aaatgccaga aggaatcaga 2341 gcagaaccgg gagaagcagc agcgtattga gaccttggag cgctacctgg ctgacctgcc 2401 cacactggaa gaccatcaga agcagagcca gcagcttaag gattctgagt tgaagagcac 2461 agagctgcag gagaaagtga ctgagctgga gagtttgctg gaggagaccc aggcaatctg 2521 cagagagaag gagattcaac tggaaagcct gaggcagaga gaagcagaat tctcctccgc 2581 tggacatagc ctgcaagata aacagtctgt ggaggagacc agtggagaag gtccagaagt 2641 ggaaatggag tcctggcaga agcgatacga ttcgctccaa aagattgtgg agaagcagca 2701 gcagaagatg gatcagttgc gctcacaagt acagagccta gagcaggaag tggctcaaga 2761 agaaggaaca agccaggccc tgagagagga ggcccagcga agggattcag ccctgcagca 2821 gctgcgcaca gccgtgaagg agctttcagt gcaaaaccag gacttgattg agaagaatct 2881 gacactccag gaacacctgc gccaggccca accagggtct ccaccttcac cagacacggc 2941 ccagctggca cttgagctgc accaggagtt ggccagttgc cttcaagatc tgcaggctgt 3001 ctgtagcatt gtgacccaga gggcccaggg ccatgacccc aatctctccc tgctcctggg 3061 cattcactca gcacagcacc cagagactca gctagatttg cagaagccag atgtgatcaa 3121 gaggaaacta gaagaggttc aacagctgcg tcgtgacatt gaggacttaa ggaccaccat 3181 gtcagacaga tatgcccagg acatgggaga aaactgtgtc acacagctcg aggactacaa 3241 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta 3301 gtgagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc 3361 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 3421 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 3481 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg 3541 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta 3601 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 3661 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 3721 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 3781 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 3841 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 3901 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 3961 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 4021 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 4081 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 4141 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 4201 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 4261 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 4321 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 4381 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt 4441 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 4501 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 4561 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 4621 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 4681 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 4741 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 4801 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 4861 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 4921 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 4981 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 5041 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 5101 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 5161 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 5221 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca 5281 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa 5341 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct 5401 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 5461 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 5521 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat 5581 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 5641 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 5701 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 5761 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 5821 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 5881 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 5941 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 6001 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 6061 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 6121 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 6181 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 6241 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 6301 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 6361 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 6421 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 6481 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc 6541 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 6601 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 6661 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 6721 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 6781 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 6841 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 6901 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 6961 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 7021 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 7081 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 7141 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 7201 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 7261 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 7321 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 7381 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 7441 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 7501 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 7561 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 7621 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 7681 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc //