LOCUS V047425 6625 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V047425 VERSION V047425 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..2376 /label="Krt14(NM_016958)" /note="Krt14(NM_016958)" /gene="Krt14" CDS 2389..2454 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2489..2713 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2759..3187 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3201..3530 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3542..4141 /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 4301..4422 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4471..4487) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4495..4511 /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(4519..4549) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4564..4585 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4873..5458) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5629..6489) /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(6490..6594) /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 acctgcagcc gccagttcac ctcctccagc 961 tccatgaagg gctcttgtgg tatcggtggt ggctctagcc gcatgtcctc catcctggct 1021 ggaggatcct gccgggctcc cagcacctac gggggcatgt cggttacctc ctctcgcttc 1081 tcctctgggg gagcctgtgg gattggaggt ggctatggtg ggagcttcag cagcagcagt 1141 tttggtggag gacttggtag tggatttggt ggtcgatttg atggatttgg tggtggtttt 1201 ggtggtggtc ttggtggtgg ttttggtggt ggtcttggtg gtggtcttgg cggtggtatt 1261 ggtgatgggc tcctggtggg cagtgagaaa gtgaccatgc agaacctcaa tgaccgcctg 1321 gccacctacc tggacaaggt gcgtgccctg gaagaggcca acactgaact ggaggtgaag 1381 attcgggact ggtaccagag gcagcggccc actgagatca aagactacag cccctacttc 1441 aagaccatcg aggacctgaa gagcaagatc ctggcagcca ccgtggacaa tgccaatgtc 1501 ctcctgcaga tcgacaatgc ccgcctggct gccgatgact tccggaccaa gtttgagacg 1561 gagcagagcc tgcgcatgag cgtggaggcc gacatcaatg gcctgcgcag ggtgctggat 1621 gagctgaccc tggccagagc cgacctggag atgcagattg agagcctcaa ggaggagctg 1681 gcctacctga agaagaacca cgaggaggaa atggcctcca tgagaggcca ggtgggtgga 1741 gacgtcaatg tggagatgga cgccgcccct ggtgtggacc tgagccgcat cctgaacgag 1801 atgcgggatc agtacgagaa gatggcggag aagaaccgca aggatgctga ggaatggttc 1861 ttcagcaaga cagaggagct gaaccgcgag gtggccacca acagcgagct ggtgcagagc 1921 ggcaagagtg agatttctga gctccggcgc accatgcaga acctggagat cgagctgcag 1981 tcccagctca gcatgaaagc atccctggag aataacctgg aggagaccaa aggccgttac 2041 tgcatgcagc tggcccagat ccaggagatg atcggcagtg tggaggagca gctggctcag 2101 ctgcgctgcg agatggagca gcagaaccag gagtacaaaa tcctgctgga tgtgaagaca 2161 aggctggagc aggagatcgc cacctaccgc cgtctgctgg agggagagga cgcccacctt 2221 tcatcttccc aattctcctc atcctctcaa ttctcctctg gctctcagtc atccagagat 2281 gtgacctcca ccaaccgcca gatccgcacc aaggtcatgg atgtgcacga tggcaaggtg 2341 gtctccaccc acgagcaggt cctgcgcacc aagaacctcg agaccggtga ctacaaagac 2401 catgacggtg attataaaga tcatgacatc gattacaagg atgacgatga caagtgaccg 2461 gtaagtttaa accgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt 2521 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa 2581 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg 2641 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg 2701 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac 2761 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct 2821 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg 2881 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt 2941 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca 3001 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga 3061 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa 3121 gggattttgg ccatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac 3181 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag 3241 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 3301 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 3361 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 3421 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc 3481 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg 3541 gatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt 3601 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga 3661 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga 3721 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga 3781 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg 3841 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga 3901 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg 3961 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct 4021 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc 4081 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg 4141 attcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc 4201 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct 4261 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta 4321 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 4381 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc 4441 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 4501 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 4561 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 4621 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 4681 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 4741 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 4801 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 4861 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 4921 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 4981 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 5041 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 5101 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 5161 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 5221 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 5281 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 5341 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 5401 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 5461 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 5521 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 5581 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 5641 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 5701 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 5761 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 5821 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 5881 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 5941 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 6001 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 6061 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 6121 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 6181 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 6241 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 6301 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 6361 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 6421 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 6481 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 6541 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 6601 tccccgaaaa gtgccacctg acgtc //