LOCUS V035034 5536 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035034 VERSION V035034 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 925..1287 /label="NMB(NM_021077)" /note="NMB(NM_021077)" /gene="NMB" CDS 1300..1365 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1400..1624 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1670..2098 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2112..2441 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2453..3052 /codon_start=1 /gene="pac from Streptomyces alboniger" /note="confers resistance to puromycin" /product="puromycin N-acetyltransferase" /transl_table=1 /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*" /label="PuroR" polyA_signal 3212..3333 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3382..3398) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3406..3422 /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(3430..3460) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3475..3496 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(3784..4369) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4540..5400) /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(5401..5505) /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 aagctttcta gagaattcgc caccatggcc cggcgggcgg ggggcgctcg gatgttcggc 961 agcctcctgc tcttcgccct gctcgctgcc ggcgtcgccc cgctcagctg ggatctcccg 1021 gagccccgca gccgagccag caagatccga gtgcactcgc gaggcaacct ctgggccacc 1081 ggtcacttca tgggcaagaa gagtctggag ccttccagcc catccccatt ggggacagct 1141 ccccacacct ccctgaggga ccagcgactg cagctgagtc atgatctgct cggaatcctc 1201 ctgctaaaga aggctctggg cgtgagcctc agccgccccg caccccaaat ccagtacagg 1261 aggctgctgg tacaaatact gcagaaactc gagaccggtg actacaaaga ccatgacggt 1321 gattataaag atcatgacat cgattacaag gatgacgatg acaagtgacc ggtaagttta 1381 aaccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc 1441 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag 1501 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag 1561 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct 1621 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt 1681 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc 1741 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc 1801 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg 1861 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga 1921 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc 1981 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg 2041 gccatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa 2101 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa 2161 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc 2221 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc 2281 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct 2341 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga 2401 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggatgaccga 2461 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct 2521 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat 2581 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 2641 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 2701 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 2761 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 2821 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 2881 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 2941 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 3001 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gattcgaaat 3061 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta 3121 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg 3181 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta 3241 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 3301 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag 3361 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 3421 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 3481 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 3541 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 3601 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 3661 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 3721 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 3781 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 3841 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 3901 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 3961 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 4021 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 4081 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 4141 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 4201 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 4261 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 4321 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 4381 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 4441 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 4501 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 4561 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 4621 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 4681 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 4741 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 4801 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 4861 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 4921 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 4981 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 5041 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 5101 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 5161 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 5221 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 5281 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 5341 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 5401 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 5461 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 5521 agtgccacct gacgtc //