LOCUS V045157 6781 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045157 VERSION V045157 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..2532 /label="HSF2(NM_004506)" /note="HSF2(NM_004506)" /gene="HSF2" CDS 2545..2610 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2645..2869 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2915..3343 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3357..3686 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3698..4297 /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 4457..4578 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4627..4643) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4651..4667 /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(4675..4705) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4720..4741 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5029..5614) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5785..6645) /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(6646..6750) /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 caccatgaag cagagttcga acgtgccggc tttcctcagc 961 aagctgtgga cgcttgtgga ggaaacccac actaacgagt tcatcacctg gagccagaat 1021 ggccaaagtt ttctggtctt ggatgagcaa cgatttgcaa aagaaattct tcccaaatat 1081 ttcaagcaca ataatatggc aagctttgtg aggcaactga atatgtatgg tttccgtaaa 1141 gtagtacata tcgactctgg aattgtaaag caagaaagag atggtcctgt agaatttcag 1201 catccttact tcaaacaagg acaggatgac ttgttggaga acattaaaag gaaggtttca 1261 tcttcaaaac cagaagaaaa taaaattcgt caggaagatt taacaaaaat tataagtagt 1321 gctcagaagg ttcagataaa acaggaaact attgagtcca ggctttctga attaaaaagt 1381 gagaatgagt ccctttggaa ggaggtgtca gaattacgag caaagcatgc acaacagcaa 1441 caagttattc gaaagattgt ccagtttatt gttacattgg ttcaaaataa ccaacttgtg 1501 agtttaaaac gtaaaaggcc tctacttcta aacactaatg gagcccaaaa gaagaacctg 1561 tttcagcaca tagtcaaaga accaactgat aatcatcatc ataaagttcc acacagtagg 1621 actgaaggtt taaagccaag ggagaggatt tcagatgaca tcattattta tgatgttact 1681 gatgataatg cagatgaaga aaatatccca gttattccag aaactaatga ggatgttata 1741 tctgatccct ccaactgtag ccagtaccct gatattgtca tcgttgaaga tgacaatgaa 1801 gatgagtatg cacctgtcat tcagagtgga gagcagaatg aaccagccag agaatcccta 1861 agttcaggca gtgatggcag cagccctctc atgtctagtg ctgtccagct aaatggctca 1921 tccagtctga cctcagaaga tccagtgacc atgatggatt ccattttgaa tgataacatc 1981 aatcttttgg gaaaggttga gctgttggat tatcttgaca gtattgactg cagtttagag 2041 gacttccagg ccatgctatc aggaagacaa tttagcatag acccagatct cctggttgat 2101 cttttcacta gttctgtgca gatgaatccc acagattaca tcaataatac aaaatctgag 2161 aataaaggat tagaaactac caagaacaat gtagttcagc cagtttcgga agagggaaga 2221 aaatctaaat ccaaaccaga taagcagctt atccagtata ccgcctttcc acttcttgca 2281 ttcctcgatg ggaaccctgc ttcttctgtt gaacaggcga gtacaacagc atcatcagaa 2341 gttttgtcct ctgtagataa acccatagaa gttgatgagc ttctggatag cagcctagac 2401 ccagaaccaa cccaaagtaa gcttgttcgc ctggagccat tgactgaagc tgaagctagt 2461 gaagctacac tgttttattt atgtgaactt gctcctgcac ctctggatag tgatatgcca 2521 cttttagata gcctcgagac cggtgactac aaagaccatg acggtgatta taaagatcat 2581 gacatcgatt acaaggatga cgatgacaag tgaccggtaa gtttaaaccg ctgatcagcc 2641 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg 2701 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat 2761 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag 2821 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg 2881 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc 2941 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 3001 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 3061 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 3121 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 3181 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 3241 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggccat ttcggcctat 3301 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt 3361 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 3421 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 3481 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 3541 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 3601 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct 3661 tttttggagg cctaggcttt tgcaaaaagc tcccgggatg accgagtaca agcccacggt 3721 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc 3781 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga 3841 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga 3901 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt 3961 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca 4021 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt 4081 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt 4141 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct 4201 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc 4261 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgattc gaaatgaccg accaagcgac 4321 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt 4381 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga 4441 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag 4501 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 4561 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta 4621 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 4681 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 4741 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 4801 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 4861 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 4921 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 4981 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 5041 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 5101 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 5161 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 5221 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 5281 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 5341 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 5401 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 5461 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 5521 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa 5581 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 5641 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 5701 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 5761 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 5821 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 5881 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 5941 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 6001 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 6061 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 6121 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 6181 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 6241 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 6301 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 6361 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 6421 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 6481 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 6541 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 6601 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 6661 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 6721 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 6781 c //