LOCUS V020337 9551 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020337 VERSION V020337 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 941..5041 /label="IGF1R(NM_000875)" /note="IGF1R(NM_000875)" /gene="IGF1R" CDS 5048..5113 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 5151..5375 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 5421..5849 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 5863..6192 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 6259..7053 /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 7227..7348 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(7397..7413) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7421..7437 /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(7445..7475) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7490..7511 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7799..8384) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8555..9415) /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(9416..9520) /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 atccgccacc atgaagtctg gctccggagg 961 agggtccccg acctcgctgt gggggctcct gtttctctcc gccgcgctct cgctctggcc 1021 gacgagtgga gaaatctgcg ggccaggcat cgacatccgc aacgactatc agcagctgaa 1081 gcgcctggag aactgcacgg tgatcgaggg ctacctccac atcctgctca tctccaaggc 1141 cgaggactac cgcagctacc gcttccccaa gctcacggtc attaccgagt acttgctgct 1201 gttccgagtg gctggcctcg agagcctcgg agacctcttc cccaacctca cggtcatccg 1261 cggctggaaa ctcttctaca actacgccct ggtcatcttc gagatgacca atctcaagga 1321 tattgggctt tacaacctga ggaacattac tcggggggcc atcaggattg agaaaaatgc 1381 tgacctctgt tacctctcca ctgtggactg gtccctgatc ctggatgcgg tgtccaataa 1441 ctacattgtg gggaataagc ccccaaagga atgtggggac ctgtgtccag ggaccatgga 1501 ggagaagccg atgtgtgaga agaccaccat caacaatgag tacaactacc gctgctggac 1561 cacaaaccgc tgccagaaaa tgtgcccaag cacgtgtggg aagcgggcgt gcaccgagaa 1621 caatgagtgc tgccaccccg agtgcctggg cagctgcagc gcgcctgaca acgacacggc 1681 ctgtgtagct tgccgccact actactatgc cggtgtctgt gtgcctgcct gcccgcccaa 1741 cacctacagg tttgagggct ggcgctgtgt ggaccgtgac ttctgcgcca acatcctcag 1801 cgccgagagc agcgactccg aggggtttgt gatccacgac ggcgagtgca tgcaggagtg 1861 cccctcgggc ttcatccgca acggcagcca gagcatgtac tgcatccctt gtgaaggtcc 1921 ttgcccgaag gtctgtgagg aagaaaagaa aacaaagacc attgattctg ttacttctgc 1981 tcagatgctc caaggatgca ccatcttcaa gggcaatttg ctcattaaca tccgacgggg 2041 gaataacatt gcttcagagc tggagaactt catggggctc atcgaggtgg tgacgggcta 2101 cgtgaagatc cgccattctc atgccttggt ctccttgtcc ttcctaaaaa accttcgcct 2161 catcctagga gaggagcagc tagaagggaa ttactccttc tacgtcctcg acaaccagaa 2221 cttgcagcaa ctgtgggact gggaccaccg caacctgacc atcaaagcag ggaaaatgta 2281 ctttgctttc aatcccaaat tatgtgtttc cgaaatttac cgcatggagg aagtgacggg 2341 gactaaaggg cgccaaagca aaggggacat aaacaccagg aacaacgggg agagagcctc 2401 ctgtgaaagt gacgtcctgc atttcacctc caccaccacg tcgaagaatc gcatcatcat 2461 aacctggcac cggtaccggc cccctgacta cagggatctc atcagcttca ccgtttacta 2521 caaggaagca ccctttaaga atgtcacaga gtatgatggg caggatgcct gcggctccaa 2581 cagctggaac atggtggacg tggacctccc gcccaacaag gacgtggagc ccggcatctt 2641 actacatggg ctgaagccct ggactcagta cgccgtttac gtcaaggctg tgaccctcac 2701 catggtggag aacgaccata tccgtggggc caagagtgag atcttgtaca ttcgcaccaa 2761 tgcttcagtt ccttccattc ccttggacgt tctttcagca tcgaactcct cttctcagtt 2821 aatcgtgaag tggaaccctc cctctctgcc caacggcaac ctgagttact acattgtgcg 2881 ctggcagcgg cagcctcagg acggctacct ttaccggcac aattactgct ccaaagacaa 2941 aatccccatc aggaagtatg ccgacggcac catcgacatt gaggaggtca cagagaaccc 3001 caagactgag gtgtgtggtg gggagaaagg gccttgctgc gcctgcccca aaactgaagc 3061 cgagaagcag gccgagaagg aggaggctga ataccgcaaa gtctttgaga atttcctgca 3121 caactccatc ttcgtgccca gacctgaaag gaagcggaga gatgtcatgc aagtggccaa 3181 caccaccatg tccagccgaa gcaggaacac cacggccgca gacacctaca acatcaccga 3241 cccggaagag ctggagacag agtacccttt ctttgagagc agagtggata acaaggagag 3301 aactgtcatt tctaaccttc ggcctttcac attgtaccgc atcgatatcc acagctgcaa 3361 ccacgaggct gagaagctgg gctgcagcgc ctccaacttc gtctttgcaa ggactatgcc 3421 cgcagaagga gcagatgaca ttcctgggcc agtgacctgg gagccaaggc ctgaaaactc 3481 catcttttta aagtggccgg aacctgagaa tcccaatgga ttgattctaa tgtatgaaat 3541 aaaatacgga tcacaagttg aggatcagcg agaatgtgtg tccagacagg aatacaggaa 3601 gtatggaggg gccaagctaa accggctaaa cccggggaac tacacagccc ggattcaggc 3661 cacatctctc tctgggaatg ggtcgtggac agatcctgtg ttcttctatg tccaggccaa 3721 aacaggatat gaaaacttca tccatctgat catcgctctg cccgtcgctg tcctgttgat 3781 cgtgggaggg ttggtgatta tgctgtacgt cttccataga aagagaaata acagcaggct 3841 ggggaatgga gtgctgtatg cctctgtgaa cccggagtac ttcagcgctg ctgatgtgta 3901 cgttcctgat gagtgggagg tggctcggga gaagatcacc atgagccggg aacttgggca 3961 ggggtcgttt gggatggtct atgaaggagt tgccaagggt gtggtgaaag atgaacctga 4021 aaccagagtg gccattaaaa cagtgaacga ggccgcaagc atgcgtgaga ggattgagtt 4081 tctcaacgaa gcttctgtga tgaaggagtt caattgtcac catgtggtgc gattgctggg 4141 tgtggtgtcc caaggccagc caacactggt catcatggaa ctgatgacac ggggcgatct 4201 caaaagttat ctccggtctc tgaggccaga aatggagaat aatccagtcc tagcacctcc 4261 aagcctgagc aagatgattc agatggccgg agagattgca gacggcatgg catacctcaa 4321 cgccaataag ttcgtccaca gagaccttgc tgcccggaat tgcatggtag ccgaagattt 4381 cacagtcaaa atcggagatt ttggtatgac gcgagatatc tatgagacag actattaccg 4441 gaaaggaggg aaagggctgc tgcccgtgcg ctggatgtct cctgagtccc tcaaggatgg 4501 agtcttcacc acttactcgg acgtctggtc cttcggggtc gtcctctggg agatcgccac 4561 actggccgag cagccctacc agggcttgtc caacgagcaa gtccttcgct tcgtcatgga 4621 gggcggcctt ctggacaagc cagacaactg tcctgacatg ctgtttgaac tgatgcgcat 4681 gtgctggcag tataacccca agatgaggcc ttccttcctg gagatcatca gcagcatcaa 4741 agaggagatg gagcctggct tccgggaggt ctccttctac tacagcgagg agaacaagct 4801 gcccgagccg gaggagctgg acctggagcc agagaacatg gagagcgtcc ccctggaccc 4861 ctcggcctcc tcgtcctccc tgccactgcc cgacagacac tcaggacaca aggccgagaa 4921 cggccccggc cctggggtgc tggtcctccg cgccagcttc gacgagagac agccttacgc 4981 ccacatgaac gggggccgca agaacgagcg ggccttgccg ctgccccagt cttcgacctg 5041 cctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga 5101 tgacgatgac aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc 5161 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc 5221 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg 5281 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa 5341 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg 5401 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt 5461 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt 5521 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct 5581 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg 5641 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga 5701 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc 5761 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga 5821 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt 5881 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 5941 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 6001 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 6061 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 6121 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 6181 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca 6241 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct 6301 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc 6361 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc 6421 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc 6481 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg 6541 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc 6601 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac 6661 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat 6721 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc 6781 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg 6841 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg 6901 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc 6961 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc 7021 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg 7081 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa 7141 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc 7201 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat 7261 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 7321 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga 7381 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 7441 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 7501 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 7561 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 7621 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 7681 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 7741 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 7801 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 7861 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 7921 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 7981 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 8041 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 8101 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 8161 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 8221 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct 8281 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt 8341 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 8401 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 8461 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 8521 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 8581 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 8641 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 8701 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 8761 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 8821 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 8881 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 8941 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 9001 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 9061 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 9121 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 9181 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 9241 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 9301 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 9361 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 9421 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 9481 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 9541 cacctgacgt c //