LOCUS V027676 5706 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027676 VERSION V027676 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" protein_bind 820..838 /bound_moiety="tetracycline repressor TetR" /gene="tetO" /note="bacterial operator O2 for the tetR and tetA genes" /label="tet operator" protein_bind 841..859 /bound_moiety="tetracycline repressor TetR" /gene="tetO" /note="bacterial operator O2 for the tetR and tetA genes" /label="tet operator" regulatory 984..993 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 993..1016 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 1023..1532 /label="ISG20(NM_001303236)" /note="ISG20(NM_001303236)" /gene="ISG20" polyA_signal 1664..1888 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" protein_bind 2172..2219 /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2μ plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." /label="FRT" CDS 2227..3249 /codon_start=1 /gene="aph(4)-Ia" /note="confers resistance to hygromycin" /product="aminoglycoside phosphotransferase from E. coli" /transl_table=1 /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE *" /label="HygR" polyA_signal 3379..3500 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3549..3565) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3573..3589 /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(3597..3627) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3642..3663 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(3951..4539) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4710..5570) /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(5571..5675) /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 ccctatcagt gatagagatc 841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga 901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga 961 ctctagcgtt taaacttaag cttgccacca tggactacaa ggacgacgat gacaaggata 1021 tcatgaaact atcaggtgtg aggagggtcc tagtgcaatc tcttcttttc aaagatgata 1081 catttgaagc ccagggagga ctgataattt gcccaaggtc acacagccag tcaaggaggg 1141 attgctccct tgccagcccc agccccaccc agaagatgct gggtcatgta ggggtgggca 1201 catgtcttcc tggccagatc ctgcagctcc tgaaaggcaa gctggtggtg ggtcatgacc 1261 tgaagcacga cttccaggca ctgaaagagg acatgagcgg ctacacaatc tacgacacgt 1321 ccactgacag gctgttgtgg cgtgaggcca agctggacca ctgcaggcgt gtctccctgc 1381 gggtgctgag tgagcgcctc ctacacaaga gcatccagaa cagcctgctt ggacacagct 1441 cggtggaaga tgcgagggca acgatggagc tctatcaaat ctcccagaga atccgagccc 1501 gccgagggct gccccgcctg gctgtgtcag actgagggga gggcatcctg tatggggagg 1561 agggggatcc actagtccag tgtggtggaa ttctgcagat atccagcaca gtggcggccg 1621 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc 1681 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc 1741 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct 1801 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg 1861 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct 1921 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 1981 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 2041 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 2101 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 2161 tcacgtacct agaagttcct attccgaagt tcctattctc tagaaagtat aggaacttcc 2221 ttggccaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc 2281 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc 2341 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa 2401 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac 2461 attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg 2521 ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg 2581 gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa 2641 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg 2701 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat 2761 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc 2821 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag 2881 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg 2941 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg 3001 ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt 3061 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc 3121 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat 3181 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca 3241 aaggaatagc acgtactacg agatttcgat tccaccgccg ccttctatga aaggttgggc 3301 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg 3361 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat 3421 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 3481 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg 3541 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac 3601 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca 3661 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat 3721 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 3781 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 3841 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 3901 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 3961 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 4021 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 4081 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 4141 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 4201 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 4261 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 4321 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 4381 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 4441 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 4501 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 4561 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 4621 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 4681 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 4741 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 4801 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 4861 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 4921 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 4981 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 5041 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 5101 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 5161 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 5221 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 5281 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 5341 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 5401 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 5461 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 5521 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 5581 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 5641 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 5701 gacgtc //