LOCUS V031837 6952 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031837 VERSION V031837 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..2703 /label="Cdyl(NM_009881)" /note="Cdyl(NM_009881)" /gene="Cdyl" CDS 2716..2781 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2816..3040 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3086..3514 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3528..3857 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3869..4468 /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 4628..4749 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4798..4814) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4822..4838 /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(4846..4876) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4891..4912 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5200..5785) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5956..6816) /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(6817..6921) /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 caccatgggc ataggcaata gccagcctaa ttcacaggaa 961 gcccagctct gcacacttcc agagaaagct gaacaaccta ctgatgataa cacctgccag 1021 caaaataatg tggttcctgc aacagtctca gaacccgatc aagcgtcccc tgcaattcaa 1081 gacgcggaga ctcaggtgga aagtatcgtt gacaaaagga aaaacaagaa agggaagaca 1141 gaatatctgg tgcggtggaa aggctatgac agtgaggatg acacgtggga gcctgagcag 1201 cacctggtga actgtgagga atacatccat gacttcaacc ggcgccacaa cgagaggcaa 1261 aaggaaggta gcctggctcg tgccagcaga gcctccccca gcaacgcccg gaagcagatt 1321 tccaggtcca cccacagcac tctctccaag accaactcca aagcacttgt ggtaggcaaa 1381 gatcatgagt ccaaaagcag ccagctgttg gctgccagcc agaagttcag gaaaaaccca 1441 gccccatctc ttgcaaaccg caagaacatg gacctcgcca agtcagggat caaaattctc 1501 gtgcctaaga gccccgttaa gggcaggacc tcggttgatg gctttcaggg ggagagcccc 1561 gagaagctgg accctgtgga tcagggtgcc gaggacactg tagccccaga ggtgactgca 1621 gagaagccca ctggggcttt gctgggccct ggtgcggagc gagccaggat ggggagcagg 1681 ccccgaatac atccactagt gcctcaggtt tctggccccg tgactgctgc catggccaca 1741 ggcttagctg ttaatggaaa aggtacatct ccattcatgg atgcgctagc agccaacgga 1801 acagtcacca tacagacatc cgtaacagga gtgacagccg ggaaaaggaa atttattgac 1861 gacagaagag accaaccttt tgacaagcgg ttgcgtttca gtgtgaggca gacagagagt 1921 gcctacagat acagagatat tgtcgtcagg aagcaagatg gcttcaccca catcttgtta 1981 tccacaaaat cgtcagagaa taactcacta aacccagagg tgatgaaaga agtgcagagc 2041 gccctgagca cagctgcagc cgacgacagc aagctggttc tgctcagcgc cgtgggcagc 2101 gtcttctgct gtggtctgga ctttatttat tttattcggc gcctcacaga tgaccgaaag 2161 agagaaagca ctaaaatggc agacgctatc agaaacttcg tgaatacttt cattcagttt 2221 aagaagccta ttattgtagc tgttaatggc ccagccattg gactaggagc atccatattg 2281 cctctttgtg atgtggtttg ggctaacgaa aaggcttggt ttcaaacacc ctataccacc 2341 ttcggacaga gtccagatgg ctgctctacc gttatgtttc ccaagattat gggaggagca 2401 tctgcgaatg aaatgctgtt cagtgggcgg aagttgacgg cacaggaggc ctgtggcaag 2461 ggtctggtct cccaggtgtt ttggccagga accttcacac aggaagtcat ggttcgaatc 2521 aaggagctgg cttcatgtaa cccagttgtc ctggaggaat ccaaagccct ggtgcgctgc 2581 aatatgaaga tggagctaga gcaggccaat gagagagaat gtgaagtgct gaagaagatc 2641 tggggctccg cccagggcat ggactccatg ttaaagtact tacagaggaa aatcgatgag 2701 ttcctcgaga ccggtgacta caaagaccat gacggtgatt ataaagatca tgacatcgat 2761 tacaaggatg acgatgacaa gtgaccggta agtttaaacc gctgatcagc ctcgactgtg 2821 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa 2881 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt 2941 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa 3001 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc 3061 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt 3121 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 3181 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 3241 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 3301 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 3361 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 3421 atctcggtct attcttttga tttataaggg attttggcca tttcggccta ttggttaaaa 3481 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag 3541 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 3601 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 3661 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 3721 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 3781 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag 3841 gcctaggctt ttgcaaaaag ctcccgggat gaccgagtac aagcccacgg tgcgcctcgc 3901 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc 3961 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga 4021 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc 4081 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat 4141 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg 4201 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc 4261 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc 4321 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta 4381 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg 4441 gtgcatgacc cgcaagcccg gtgcctgatt cgaaatgacc gaccaagcga cgcccaacct 4501 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt 4561 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc 4621 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 4681 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 4741 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc 4801 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 4861 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 4921 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 4981 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 5041 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 5101 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 5161 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 5221 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 5281 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 5341 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 5401 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 5461 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 5521 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 5581 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 5641 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 5701 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat 5761 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 5821 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 5881 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 5941 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 6001 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg 6061 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga 6121 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt 6181 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 6241 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 6301 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 6361 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc 6421 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 6481 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 6541 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 6601 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt 6661 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 6721 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 6781 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg 6841 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 6901 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc //