LOCUS V037046 6779 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V037046 VERSION V037046 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 683..1459 /label="IGFBP1(NM_000596)" /note="IGFBP1(NM_000596)" /gene="IGFBP1" CDS 1466..1531 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1555..2143 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2209..2257 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2459..2887 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2901..3230 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3297..4091 /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 4267..4388 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4437..4453) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4461..4477 /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(4485..4515) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4530..4551 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4839..5427) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5598..6458) /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(6459..6563) /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 ggatccgcca ccatgtcaga ggtccccgtt gctcgcgtct ggctggtact 721 gctcctgctg actgtccagg tcggcgtgac agccggcgct ccgtggcagt gcgcgccctg 781 ctccgccgag aagctcgcgc tctgcccgcc ggtgtccgcc tcgtgctcgg aggtcacccg 841 gtccgccggc tgcggctgtt gcccgatgtg cgccctgcct ctgggcgccg cgtgcggcgt 901 ggcgactgca cgctgcgccc ggggactcag ttgccgcgcg ctgccggggg agcagcaacc 961 tctgcacgcc ctcacccgcg gccaaggcgc ctgcgtgcag gagtctgacg cctccgctcc 1021 ccatgctgca gaggcaggga gccctgaaag cccagagagc acggagataa ctgaggagga 1081 gctcctggat aatttccatc tgatggcccc ttctgaagag gatcattcca tcctttggga 1141 cgccatcagt acctatgatg gctcgaaggc tctccatgtc accaacatca aaaaatggaa 1201 ggagccctgc cgaatagaac tctacagagt cgtagagagt ttagccaagg cacaggagac 1261 atcaggagaa gaaatttcca aattttacct gccaaactgc aacaagaatg gattttatca 1321 cagcagacag tgtgagacat ccatggatgg agaggcggga ctctgctggt gcgtctaccc 1381 ttggaatggg aagaggatcc ctgggtctcc agagatcagg ggagacccca actgccagat 1441 atattttaat gtacaaaacc tcgaggacta caaagaccat gacggtgatt ataaagatca 1501 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct cgacaatcaa 1561 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt 1621 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct 1681 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc 1741 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg 1801 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc 1861 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc 1921 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt 1981 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca 2041 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt 2101 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc 2161 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa 2221 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag 2281 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct 2341 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc 2401 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac 2461 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct 2521 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg 2581 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt 2641 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca 2701 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga 2761 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa 2821 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac 2881 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag 2941 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 3001 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 3061 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 3121 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc 3181 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg 3241 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga 3301 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct 3361 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc 3421 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg 3481 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg 3541 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc 3601 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc 3661 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg 3721 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc 3781 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg 3841 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc 3901 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag 3961 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg 4021 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg 4081 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg 4141 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt 4201 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc 4261 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 4321 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 4381 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca 4441 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga 4501 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg 4561 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 4621 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 4681 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 4741 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 4801 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 4861 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 4921 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 4981 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca 5041 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 5101 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 5161 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 5221 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg 5281 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 5341 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 5401 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 5461 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 5521 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 5581 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 5641 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 5701 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 5761 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 5821 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 5881 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 5941 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 6001 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 6061 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 6121 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 6181 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 6241 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 6301 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 6361 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 6421 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 6481 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 6541 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga 6601 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat 6661 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca 6721 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg //