LOCUS V037776 6289 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V037776 VERSION V037776 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..2040 /label="CDK9(NM_001261)" /note="CDK9(NM_001261)" /gene="CDK9" CDS 2053..2118 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2153..2377 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2423..2851 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2865..3194 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3206..3805 /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 3965..4086 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4135..4151) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4159..4175 /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(4183..4213) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4228..4249 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4537..5122) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5293..6153) /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(6154..6258) /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 caccatggca aagcagtacg actcggtgga gtgccctttt 961 tgtgatgaag tttccaaata cgagaagctc gccaagatcg gccaaggcac cttcggggag 1021 gtgttcaagg ccaggcaccg caagaccggc cagaaggtgg ctctgaagaa ggtgctgatg 1081 gaaaacgaga aggaggggtt ccccattaca gccttgcggg agatcaagat ccttcagctt 1141 ctaaaacacg agaatgtggt caacttgatt gagatttgtc gaaccaaagc ttccccctat 1201 aaccgctgca agggtagtat atacctggtg ttcgacttct gcgagcatga ccttgctggg 1261 ctgttgagca atgttttggt caagttcacg ctgtctgaga tcaagagggt gatgcagatg 1321 ctgcttaacg gcctctacta catccacaga aacaagatcc tgcataggga catgaaggct 1381 gctaatgtgc ttatcactcg tgatggggtc ctgaagctgg cagactttgg gctggcccgg 1441 gccttcagcc tggccaagaa cagccagccc aaccgctaca ccaaccgtgt ggtgacactc 1501 tggtaccggc ccccggagct gttgctcggg gagcgggact acggcccccc cattgacctg 1561 tggggtgctg ggtgcatcat ggcagagatg tggacccgca gccccatcat gcagggcaac 1621 acggagcagc accaactcgc cctcatcagt cagctctgcg gctccatcac ccctgaggtg 1681 tggccaaacg tggacaacta tgagctgtac gaaaagctgg agctggtcaa gggccagaag 1741 cggaaggtga aggacaggct gaaggcctat gtgcgtgacc catacgcact ggacctcatc 1801 gacaagctgc tggtgctgga ccctgcccag cgcatcgaca gcgatgacgc cctcaaccac 1861 gacttcttct ggtccgaccc catgccctcc gacctcaagg gcatgctctc cacccacctg 1921 acgtccatgt tcgagtactt ggcaccaccg cgccggaagg gcagccagat cacccagcag 1981 tccaccaacc agagtcgcaa tcccgccacc accaaccaga cggagtttga gcgcgtcttc 2041 ctcgagaccg gtgactacaa agaccatgac ggtgattata aagatcatga catcgattac 2101 aaggatgacg atgacaagtg accggtaagt ttaaaccgct gatcagcctc gactgtgcct 2161 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt 2221 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg 2281 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac 2341 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc 2401 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg 2461 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct 2521 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg 2581 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag 2641 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg 2701 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc 2761 tcggtctatt cttttgattt ataagggatt ttggccattt cggcctattg gttaaaaaat 2821 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt 2881 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 2941 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 3001 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc 3061 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 3121 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 3181 taggcttttg caaaaagctc ccgggatgac cgagtacaag cccacggtgc gcctcgccac 3241 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc 3301 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact 3361 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc 3421 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg 3481 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct 3541 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc 3601 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga 3661 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga 3721 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg 3781 catgacccgc aagcccggtg cctgattcga aatgaccgac caagcgacgc ccaacctgcc 3841 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt 3901 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca 3961 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 4021 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 4081 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata 4141 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 4201 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 4261 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 4321 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 4381 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 4441 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 4501 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 4561 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 4621 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 4681 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 4741 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 4801 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 4861 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 4921 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 4981 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 5041 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac 5101 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 5161 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 5221 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 5281 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 5341 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 5401 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 5461 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 5521 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 5581 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 5641 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 5701 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 5761 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 5821 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 5881 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 5941 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 6001 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 6061 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 6121 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 6181 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 6241 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc //