LOCUS V026767 6023 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026767 VERSION V026767 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..1513 /label="Cib1(NM_031145)" /note="Cib1(NM_031145)" /gene="Cib1" CDS 1520..1585 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1623..1847 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1893..2321 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2335..2664 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2731..3525 /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 3699..3820 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3869..3885) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3893..3909 /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(3917..3947) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3962..3983 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4271..4856) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5027..5887) /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(5888..5992) /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 atgggaggtt cgggcagtcg 961 cctgtctaag gagctgctgg ccgagtacca ggacctgacg ttcctgacca agcaggagat 1021 cctccttgcc cacagacgct tttgcgagtt gcttccccca gagcatcgga ccgtggaaga 1081 gtcactgcat acccgagtat cttttgagca gatcctcagc cttccagagc tcaaggccaa 1141 ccccttcaag gagcgaatct gcatggtctt ctccacgtcg cctaccagag acagcctgag 1201 ctttgaggac ttcctggacc tcctgagtgt cttcagtgac acagcaaccc cagacatcaa 1261 gtcacactat gccttccgca tctttgactt tgatgatgat ggaaccctgg acagagaaga 1321 cctgagccgg cttgtgaact gcctcacggg agagggcgag gacactcggc tcagtgcttc 1381 tgagatgaag cagctcatcg acaatatcct ggaagagtcg gacattgaca gggatgggac 1441 catcaatctc tccgagttcc agcacgtcat ctctcgctca ccagactttg ccagctcctt 1501 taagattgtc ctgctcgagg actacaaaga ccatgacggt gattataaag atcatgacat 1561 cgactacaag gatgacgatg acaagtagtg agggcccgtt taaacccgct gatcagcctc 1621 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac 1681 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg 1741 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga 1801 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga 1861 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc 1921 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc 1981 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct 2041 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa 2101 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc 2161 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact 2221 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg 2281 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt 2341 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 2401 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg 2461 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg 2521 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt 2581 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt 2641 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct 2701 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt 2761 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc 2821 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag 2881 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg 2941 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac 3001 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc 3061 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc 3121 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc 3181 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg 3241 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat 3301 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc 3361 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa 3421 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat 3481 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt 3541 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg 3601 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc 3661 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata 3721 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc 3781 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga 3841 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc 3901 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct 3961 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa 4021 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 4081 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 4141 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 4201 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 4261 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 4321 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 4381 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 4441 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 4501 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 4561 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 4621 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 4681 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga 4741 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 4801 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 4861 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 4921 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 4981 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 5041 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 5101 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 5161 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 5221 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 5281 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 5341 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 5401 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 5461 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 5521 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 5581 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 5641 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 5701 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 5761 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 5821 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 5881 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 5941 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 6001 cccgaaaagt gccacctgac gtc //