LOCUS V047655 7165 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V047655 VERSION V047655 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 964..2655 /label="Cdc7(NM_001271566)" /note="Cdc7(NM_001271566)" /gene="Cdc7" CDS 2662..2727 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2765..2989 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3035..3463 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3477..3806 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3873..4667 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 4841..4962 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5011..5027) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5035..5051 /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(5059..5089) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5104..5125 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5413..5998) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6169..7029) /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(7030..7134) /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 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc 961 accatggagg agccaatggc gttttcttcc cttcgtggca gtgacaggtg tccggctgat 1021 gactctttaa aaaaatatga gcagagtgtt aaactttcag gtattaaaag agatattgag 1081 gagctttgtg aagctgtacc acaactagtc aatgtgttca aaattaagga caaaattgga 1141 gaaggcacct tcagctctgt ttatttggcc acagcacagt tgcaagaagg acatgaagag 1201 aaaattgcgc tgaaacactt aattcccaca agtcatccta tgaggattgc agcagagctt 1261 cagtgtctga cagttgcagg ggggcaagac aatgtcatgg gacttaagta ctgcttcaga 1321 aaaaatgatc atgtggttat tgctatgccg tatctggaac atgagtcctt tttggacatt 1381 ttgaattctc tttccttcca agaagttcgg gaatatatgt ataatctttt tgtagctttg 1441 aaacggattc atcagtttgg tattgttcac cgtgatgtga agcccagcaa ttttttatac 1501 aatagacgtc tgaaaaagta tgccttggtg gacttcggtt tggcccaggg aacccgtgac 1561 acaaaaatag agctgctcaa gtttgtccag tctgaggctc agcaggaaga ttgttcacga 1621 aacaagtatc atggagtcgt tggacacaag ggcctgctga gtcgcccggc acctaaaact 1681 gtggatcagc agtgtacccc aaaaacttct gtcaaaagat cctacacaca agttcacatt 1741 aagcaaggaa aagacggaaa ggagagatct gtaggccttt ctgtccagcg ctctgttttt 1801 ggagagagaa atttcaatat acacagctcc atttcacatg agagccctgc agagaaactc 1861 atcaagcaat caaagactgt ggacataata tcacgaaagc tagcgacaaa aaagacggcc 1921 atttctacaa aagctatgaa cagcgtgatg agggaaactg ccaggtcctg ccctgctgtc 1981 ctcacctgcg actgctatgg atcagataga gtctgcagcg tttgcctgtc gaggcggcag 2041 caggttgccc ctagggcagg cacaccagga ttcagagcgc cagaggtcct gacaaagtgt 2101 cctgaccaga ccacagcgat tgacatgtgg tctgcaggtg tcatattcct gtccttgctc 2161 agtgggcggt acccatttta caaggccagt gatgacttaa ctgctttggc tcagatcatg 2221 acaattcgag gatccaggga aactatccag gctgctaaag cttttggcaa atcagttctg 2281 tgtagcaaag aagtcccagc acaagacttg agagctctct gtgagagact gcggggtcta 2341 gactctacca ctcccaggtc agccagtggt cctccaggga atgcttccta tgaccctgct 2401 gcttccaaga acactgacca caaagcatcc cgtgtacagg ctgctcaggc acagcactca 2461 gaggactcct tgtataaaag ggacaacgat ggctattgga gtcatcccaa agactgcact 2521 tccaactcag aaggctggga ctcagtacct gatgaagcct atgacctgct cgacaagctt 2581 ctggacctaa acccagcttc aaggataaca gcagaagcag ccttattaca tgcgttcttt 2641 aaagatatgt gctccctcga ggactacaaa gaccatgacg gtgattataa agatcatgac 2701 atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg ctgatcagcc 2761 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg 2821 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat 2881 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag 2941 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg 3001 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc 3061 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 3121 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 3181 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 3241 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 3301 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 3361 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat 3421 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt 3481 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 3541 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 3601 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 3661 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 3721 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct 3781 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat 3841 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag 3901 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg 3961 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca 4021 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc 4081 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg 4141 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg 4201 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta 4261 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag 4321 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac 4381 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg 4441 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg 4501 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg 4561 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg 4621 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg 4681 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc 4741 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct 4801 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta 4861 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 4921 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc 4981 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 5041 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 5101 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 5161 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 5221 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 5281 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 5341 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 5401 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 5461 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 5521 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 5581 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 5641 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 5701 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 5761 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 5821 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 5881 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 5941 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 6001 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 6061 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 6121 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 6181 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 6241 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 6301 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 6361 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 6421 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 6481 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 6541 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 6601 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 6661 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 6721 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 6781 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 6841 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 6901 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 6961 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 7021 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 7081 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 7141 tccccgaaaa gtgccacctg acgtc //