LOCUS V027307 10080 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027307 VERSION V027307 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 706..4806 /label="IGF1R(NM_000875)" /note="IGF1R(NM_000875)" /gene="IGF1R" CDS 4813..4836 /codon_start=1 /product="8xHis affinity tag" /transl_table=1 /translation="HHHHHHHH" /label="8xHis" misc_feature 4856..5444 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 5510..5558 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 5760..6188 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 6202..6531 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 6598..7392 /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 7568..7689 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(7738..7754) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7762..7778 /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(7786..7816) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7831..7852 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8140..8728) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8899..9759) /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(9760..9864) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg 661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgaa gtctggctcc 721 ggaggagggt ccccgacctc gctgtggggg ctcctgtttc tctccgccgc gctctcgctc 781 tggccgacga gtggagaaat ctgcgggcca ggcatcgaca tccgcaacga ctatcagcag 841 ctgaagcgcc tggagaactg cacggtgatc gagggctacc tccacatcct gctcatctcc 901 aaggccgagg actaccgcag ctaccgcttc cccaagctca cggtcattac cgagtacttg 961 ctgctgttcc gagtggctgg cctcgagagc ctcggagacc tcttccccaa cctcacggtc 1021 atccgcggct ggaaactctt ctacaactac gccctggtca tcttcgagat gaccaatctc 1081 aaggatattg ggctttacaa cctgaggaac attactcggg gggccatcag gattgagaaa 1141 aatgctgacc tctgttacct ctccactgtg gactggtccc tgatcctgga tgcggtgtcc 1201 aataactaca ttgtggggaa taagccccca aaggaatgtg gggacctgtg tccagggacc 1261 atggaggaga agccgatgtg tgagaagacc accatcaaca atgagtacaa ctaccgctgc 1321 tggaccacaa accgctgcca gaaaatgtgc ccaagcacgt gtgggaagcg ggcgtgcacc 1381 gagaacaatg agtgctgcca ccccgagtgc ctgggcagct gcagcgcgcc tgacaacgac 1441 acggcctgtg tagcttgccg ccactactac tatgccggtg tctgtgtgcc tgcctgcccg 1501 cccaacacct acaggtttga gggctggcgc tgtgtggacc gtgacttctg cgccaacatc 1561 ctcagcgccg agagcagcga ctccgagggg tttgtgatcc acgacggcga gtgcatgcag 1621 gagtgcccct cgggcttcat ccgcaacggc agccagagca tgtactgcat cccttgtgaa 1681 ggtccttgcc cgaaggtctg tgaggaagaa aagaaaacaa agaccattga ttctgttact 1741 tctgctcaga tgctccaagg atgcaccatc ttcaagggca atttgctcat taacatccga 1801 cgggggaata acattgcttc agagctggag aacttcatgg ggctcatcga ggtggtgacg 1861 ggctacgtga agatccgcca ttctcatgcc ttggtctcct tgtccttcct aaaaaacctt 1921 cgcctcatcc taggagagga gcagctagaa gggaattact ccttctacgt cctcgacaac 1981 cagaacttgc agcaactgtg ggactgggac caccgcaacc tgaccatcaa agcagggaaa 2041 atgtactttg ctttcaatcc caaattatgt gtttccgaaa tttaccgcat ggaggaagtg 2101 acggggacta aagggcgcca aagcaaaggg gacataaaca ccaggaacaa cggggagaga 2161 gcctcctgtg aaagtgacgt cctgcatttc acctccacca ccacgtcgaa gaatcgcatc 2221 atcataacct ggcaccggta ccggccccct gactacaggg atctcatcag cttcaccgtt 2281 tactacaagg aagcaccctt taagaatgtc acagagtatg atgggcagga tgcctgcggc 2341 tccaacagct ggaacatggt ggacgtggac ctcccgccca acaaggacgt ggagcccggc 2401 atcttactac atgggctgaa gccctggact cagtacgccg tttacgtcaa ggctgtgacc 2461 ctcaccatgg tggagaacga ccatatccgt ggggccaaga gtgagatctt gtacattcgc 2521 accaatgctt cagttccttc cattcccttg gacgttcttt cagcatcgaa ctcctcttct 2581 cagttaatcg tgaagtggaa ccctccctct ctgcccaacg gcaacctgag ttactacatt 2641 gtgcgctggc agcggcagcc tcaggacggc tacctttacc ggcacaatta ctgctccaaa 2701 gacaaaatcc ccatcaggaa gtatgccgac ggcaccatcg acattgagga ggtcacagag 2761 aaccccaaga ctgaggtgtg tggtggggag aaagggcctt gctgcgcctg ccccaaaact 2821 gaagccgaga agcaggccga gaaggaggag gctgaatacc gcaaagtctt tgagaatttc 2881 ctgcacaact ccatcttcgt gcccagacct gaaaggaagc ggagagatgt catgcaagtg 2941 gccaacacca ccatgtccag ccgaagcagg aacaccacgg ccgcagacac ctacaacatc 3001 accgacccgg aagagctgga gacagagtac cctttctttg agagcagagt ggataacaag 3061 gagagaactg tcatttctaa ccttcggcct ttcacattgt accgcatcga tatccacagc 3121 tgcaaccacg aggctgagaa gctgggctgc agcgcctcca acttcgtctt tgcaaggact 3181 atgcccgcag aaggagcaga tgacattcct gggccagtga cctgggagcc aaggcctgaa 3241 aactccatct ttttaaagtg gccggaacct gagaatccca atggattgat tctaatgtat 3301 gaaataaaat acggatcaca agttgaggat cagcgagaat gtgtgtccag acaggaatac 3361 aggaagtatg gaggggccaa gctaaaccgg ctaaacccgg ggaactacac agcccggatt 3421 caggccacat ctctctctgg gaatgggtcg tggacagatc ctgtgttctt ctatgtccag 3481 gccaaaacag gatatgaaaa cttcatccat ctgatcatcg ctctgcccgt cgctgtcctg 3541 ttgatcgtgg gagggttggt gattatgctg tacgtcttcc atagaaagag aaataacagc 3601 aggctgggga atggagtgct gtatgcctct gtgaacccgg agtacttcag cgctgctgat 3661 gtgtacgttc ctgatgagtg ggaggtggct cgggagaaga tcaccatgag ccgggaactt 3721 gggcaggggt cgtttgggat ggtctatgaa ggagttgcca agggtgtggt gaaagatgaa 3781 cctgaaacca gagtggccat taaaacagtg aacgaggccg caagcatgcg tgagaggatt 3841 gagtttctca acgaagcttc tgtgatgaag gagttcaatt gtcaccatgt ggtgcgattg 3901 ctgggtgtgg tgtcccaagg ccagccaaca ctggtcatca tggaactgat gacacggggc 3961 gatctcaaaa gttatctccg gtctctgagg ccagaaatgg agaataatcc agtcctagca 4021 cctccaagcc tgagcaagat gattcagatg gccggagaga ttgcagacgg catggcatac 4081 ctcaacgcca ataagttcgt ccacagagac cttgctgccc ggaattgcat ggtagccgaa 4141 gatttcacag tcaaaatcgg agattttggt atgacgcgag atatctatga gacagactat 4201 taccggaaag gagggaaagg gctgctgccc gtgcgctgga tgtctcctga gtccctcaag 4261 gatggagtct tcaccactta ctcggacgtc tggtccttcg gggtcgtcct ctgggagatc 4321 gccacactgg ccgagcagcc ctaccagggc ttgtccaacg agcaagtcct tcgcttcgtc 4381 atggagggcg gccttctgga caagccagac aactgtcctg acatgctgtt tgaactgatg 4441 cgcatgtgct ggcagtataa ccccaagatg aggccttcct tcctggagat catcagcagc 4501 atcaaagagg agatggagcc tggcttccgg gaggtctcct tctactacag cgaggagaac 4561 aagctgcccg agccggagga gctggacctg gagccagaga acatggagag cgtccccctg 4621 gacccctcgg cctcctcgtc ctccctgcca ctgcccgaca gacactcagg acacaaggcc 4681 gagaacggcc ccggccctgg ggtgctggtc ctccgcgcca gcttcgacga gagacagcct 4741 tacgcccaca tgaacggggg ccgcaagaac gagcgggcct tgccgctgcc ccagtcttcg 4801 acctgcctcg agcaccatca ccatcaccat caccattagg gcccgatatc tcgacaatca 4861 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt 4921 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc 4981 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc 5041 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg 5101 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc 5161 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg 5221 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg 5281 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc 5341 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct 5401 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg 5461 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa 5521 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca 5581 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc 5641 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt 5701 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca 5761 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 5821 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 5881 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag 5941 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc 6001 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg 6061 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata 6121 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa 6181 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca 6241 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc 6301 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata 6361 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 6421 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag 6481 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg 6541 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg 6601 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc 6661 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg 6721 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag 6781 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc 6841 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat 6901 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg 6961 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc 7021 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag 7081 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc 7141 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc 7201 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata 7261 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc 7321 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac 7381 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct 7441 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt 7501 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc 7561 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 7621 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 7681 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc 7741 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 7801 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 7861 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 7921 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 7981 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 8041 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 8101 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 8161 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 8221 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 8281 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc 8341 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 8401 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 8461 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 8521 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 8581 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 8641 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 8701 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 8761 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 8821 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 8881 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 8941 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 9001 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 9061 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 9121 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 9181 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 9241 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 9301 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 9361 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 9421 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 9481 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 9541 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 9601 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 9661 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 9721 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 9781 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 9841 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg 9901 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca 9961 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc 10021 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg //