LOCUS V023193 6467 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023193 VERSION V023193 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..1957 /label="Ciao1(NM_001008766)" /note="Ciao1(NM_001008766)" /gene="Ciao1" CDS 1964..2029 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2067..2291 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2337..2765 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2779..3108 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3175..3969 /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 4143..4264 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4313..4329) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4337..4353 /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(4361..4391) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4406..4427 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4715..5300) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5471..6331) /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(6332..6436) /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 atgaaggatg cgctggtact 961 gcagagccgc gtcccggcgc acccggattc ccgctgctgg ttcctggcct ggaaccccac 1021 gggaaccttg ttggcctcat gcggaggcga ccgtaaaatt cgcatctggg gcaccgaggg 1081 tgacagctgg atttgcaagt ctgtcctttc tgaaggccat cagcgcaccg ttaggaaagt 1141 ggcctggtcc ccctgtggaa attacctggc atctgccagc tttgatgcta ctacttgcat 1201 ttggaagaag aaccaggatg actttgagtg tgtgaccact cttgagggtc atgagaatga 1261 ggtcaagtca gtggcttggg ccccatctgg caacctcttg gctacctgca gcagagataa 1321 gagcgtgtgg gtctgggaag ttgatgaaga agatgagtat gaatgcgtca gtgtcctcaa 1381 ttctcacaca caggatgtta agcatgtggt ttggcaccca agccaggagc ttttagcttc 1441 tgctagctat gatgacacag tgaagctata ccaggaagaa ggggatgact gggtctgctg 1501 tgctaccctt gaaggtcacg agtccactgt atggagcata gcctttgatc ccagcggcca 1561 gcgtctggca tcttgcagtg atgaccgaac tgtgcgcatc tggcgccagt atctacctgg 1621 caatgaacaa ggggtggcat gcagtggctc cgaccccagc tggaaatgtg tttgcacttt 1681 gtcgggcttc cataccagga ccatctacga cgttgcttgg tgtcagctga caggggccct 1741 ggctacagct tgtggagatg atgccatccg agtgtttgag gaagatccag gctcagaccc 1801 acagcagccc accttctctt tgacagccca tcttcgtcag gcccattccc aggatgtcaa 1861 ctgtgtggct tggaacccca aggaggcagg gctcctggcc tcttgtagtg atgatgggga 1921 ggtagccttc tgggagtatc atcagcctgc aggtctcctc gaggactaca aagaccatga 1981 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc 2041 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg 2101 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 2161 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 2221 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt 2281 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc 2341 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 2401 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 2461 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 2521 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc 2581 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 2641 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg 2701 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 2761 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca 2821 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca 2881 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc 2941 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 3001 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta 3061 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga 3121 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt 3181 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat 3241 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag 3301 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac 3361 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac 3421 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc 3481 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg 3541 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag 3601 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat 3661 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag 3721 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc 3781 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg 3841 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg 3901 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag 3961 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat 4021 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 4081 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 4141 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 4201 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 4261 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 4321 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 4381 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 4441 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 4501 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 4561 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 4621 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 4681 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 4741 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 4801 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 4861 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 4921 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 4981 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 5041 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 5101 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 5161 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 5221 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc 5281 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 5341 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 5401 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 5461 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 5521 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 5581 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 5641 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 5701 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 5761 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 5821 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 5881 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 5941 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 6001 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 6061 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 6121 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 6181 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 6241 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 6301 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 6361 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 6421 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc //