LOCUS 62056_8025 4157 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4157) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..4157 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 52..1036 /label=CMV IE94 promoter /note="enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94" promoter 1071..1089 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" CDS 1136..1201 /codon_start=1 /label=3xFLAG /note="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /translation="DYKDHDGDYKDHDIDYKDDDDK" polyA_signal complement(1242..1376) /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter complement(1489..1507) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(1528..1544) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(1552..1568) /label=lac operator /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-beta-D-thiogalactopyranoside (IPTG)." promoter complement(1576..1606) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(1621..1642) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(1930..2518) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2692..3549) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(3550..3654) /label=AmpR promoter rep_origin complement(3680..4135) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" ORIGIN 1 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt cgaccatagc 61 caattcaata tggcgtatat ggactcatgc caattcaata tggtggatct ggacctgtgc 121 caattcaata tggcgtatat ggactcgtgc caattcaata tggtggatct ggaccccagc 181 caattcaata tggcggactt ggcaccatgc caattcaata tggcggactt ggcactgtgc 241 caactgggga ggggtctact tggcacggtg ccaagtttga ggaggggtct tggccctgtg 301 ccaagtccgc catattgaat tggcatggtg ccaataatgg cggccatatt ggctatatgc 361 caggatcaat atataggcaa tatccaatat ggccctatgc caatatggct attggccagg 421 ttcaatacta tgtattggcc ctatgccata tagtattcca tatatgggtt ttcctattga 481 cgtagatagc ccctcccaat gggcggtccc atataccata tatggggctt cctaataccg 541 cccatagcca ctcccccatt gacgtcaatg gtctctatat atggtctttc ctattgacgt 601 catatgggcg gtcctattga cgtatatggc gcctccccca ttgacgtcaa ttacggtaaa 661 tggcccgcct ggctcaatgc ccattgacgt caataggacc acccaccatt gacgtcaatg 721 ggatggctca ttgcccattc atatccgttc tcacgccccc tattgacgtc aatgacggta 781 aatggcccac ttggcagtac atcaatatct attaatagta acttggcaag tacattacta 841 ttggaaggac gccagggtac attggcagta ctcccattga cgtcaatggc ggtaaatggc 901 ccgcgatggc tgccaagtac atccccattg acgtcaatgg ggaggggcaa tgacgcaaat 961 gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa tgggaggtct atataagcaa 1021 tgctcgttta gggaaccgcc attctgcctg gggacgtcgg agcaagcttg atttaggtga 1081 cactatagaa tacaagctac ttgttctttt tgcaggatcc catcgattcg aattcgacta 1141 caaagaccat gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa 1201 gtgactcgag cctctagaac tatagtgagt cgtattacgt agatccagac atgataagat 1261 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg 1321 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca 1381 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaat 1441 tcgcggccgc ggcgccaatg cattgggccc ggtacccagc ttttgttccc tttagtgagg 1501 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 1561 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 1621 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 1681 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 1741 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 1801 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 1861 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 1921 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 1981 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 2041 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 2101 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 2161 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 2221 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 2281 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 2341 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 2401 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 2461 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 2521 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 2581 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 2641 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 2701 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 2761 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 2821 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 2881 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 2941 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 3001 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 3061 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 3121 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 3181 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 3241 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 3301 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 3361 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 3421 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 3481 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 3541 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 3601 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 3661 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat 3721 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa 3781 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta 3841 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca 3901 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat 3961 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg 4021 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc 4081 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat 4141 tcgccattca ggctgcg //