LOCUS V031531 8045 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031531 VERSION V031531 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers primer_bind 141..157 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" enhancer 318..697 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 698..901 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 945..968 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 987..2774 /label="TCF7L2(NM_030756)" /note="TCF7L2(NM_030756)" /gene="TCF7L2" polyA_signal 2804..3426 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" promoter 3468..3765 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3819..4613 /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 5265..5399 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" promoter complement(5405..5423) /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" primer_bind complement(5437..5453) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5461..5477 /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(5485..5515) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5530..5551 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5839..6427) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6598..7458) /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(7459..7563) /gene="bla" /label="AmpR promoter" rep_origin 7590..8018 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" ORIGIN 1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa 181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta 241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc 301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg 361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga 601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg 661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac 721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt 781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc 841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc 901 tcgtttagtg aaccgtcaga attaattcag atcgatctac catggactac aaagacgatg 961 acgacaagct tgcggccgcg aattctatgc cgcagctgaa cggcggtgga ggggatgacc 1021 taggcgccaa cgacgaactg atttccttca aagacgaggg cgaacaggag gagaagagct 1081 ccgaaaactc ctcggcagag agggatttag ctgatgtcaa atcgtctcta gtcaatgaat 1141 cagaaacgaa tcaaaacagc tcctccgatt ccgaggcgga aagacggcct ccgcctcgct 1201 ccgaaagttt ccgagacaaa tcccgggaaa gtttggaaga agcggccaag aggcaagatg 1261 gagggctctt taaggggcca ccgtatcccg gctacccctt catcatgatc cccgacctga 1321 cgagccccta cctccccaac ggatcgctct cgcccaccgc ccgaacctat ctccagatga 1381 aatggccact gcttgatgtc caggcaggga gcctccagag tagacaagcc ctcaaggatg 1441 cccggtcccc atcaccggca cacattgtct ctaacaaagt gccagtggtg cagcaccctc 1501 accatgtcca ccccctcacg cctcttatca cgtacagcaa tgaacacttc acgccgggaa 1561 acccacctcc acacttacca gccgacgtag accccaaaac aggaatccca cggcctccgc 1621 accctccaga tatatccccg tattacccac tatcgcctgg caccgtagga caaatccccc 1681 atccgctagg atggttagta ccacagcaag gtcaaccagt gtacccaatc acgacaggag 1741 gattcagaca cccctacccc acagctctga ccgtcaatgc ttccatgtcc aggttccctc 1801 cccatatggt cccaccacat catacgctac acacgacggg cattccgcat ccggccatag 1861 tcacaccaac agtcaaacag gaatcgtccc agagtgatgt cggctcactc catagttcaa 1921 agcatcagga ctccaaaaag gaagaagaaa agaagaagcc ccacataaag aaacctctta 1981 atgcattcat gttgtatatg aaggaaatga gagcaaaggt cgtagctgag tgcacgttga 2041 aagaaagcgc ggccatcaac cagatccttg ggcggaggtg gcatgcactg tccagagaag 2101 agcaagcgaa atactacgag ctggcccgga aggagcgaca gcttcatatg caactgtacc 2161 ccggctggtc cgcgcgggat aactatggaa agaagaagaa gaggaaaagg gacaagcagc 2221 cgggagagac caatgaacac agcgaatgtt tcctaaatcc ttgcctttca cttcctccga 2281 ttacagacct gagcgctcct aagaaatgcc gagcgcgctt tggccttgat caacagaata 2341 actggtgcgg cccttgcagg agaaaaaaaa agtgcgttcg ctacatacaa ggtgaaggca 2401 gctgcctcag cccaccctct tcagatggaa gcttactaga ttcgcctccc ccctccccga 2461 acctgctagg ctcccctccc cgagacgcca agtcacagac tgagcagacc cagcctctgt 2521 cgctgtccct gaagcccgac cccctggccc acctgtccat gatgcctccg ccacccgccc 2581 tcctgctcgc tgaggccacc cacaaggcct ccgccctctg tcccaacggg gccctggacc 2641 tgcccccagc cgctttgcag cctgccgccc cctcctcatc aattgcacag ccgtcgactt 2701 cttccttaca ttcccacagc tccctggccg ggacccagcc ccagccgctg tcgctcgtca 2761 ccaagtcttt agaatagggt accagtcgac tctagaggat cccgggtggc atccctgtga 2821 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt 2881 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg 2941 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg 3001 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc 3061 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg 3121 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc 3181 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct 3241 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa aataactata 3301 ccagcaggag gacgtccaga cacagcatag gctacctggc catgcccaac cggtgggaca 3361 tttgagttgc ttgcttggca ctgtcctctc atgcgttggg tccactcagt agatgcctgt 3421 tgaattgggt acgcggccag cttggctgtg gaatgtgtgt cagttagggt gtggaaagtc 3481 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag 3541 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta 3601 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc 3661 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 3721 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggaa ttgatcagct 3781 tgggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc 3841 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga 3901 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt 3961 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat 4021 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg 4081 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg 4141 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc 4201 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga 4261 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag 4321 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc 4381 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg 4441 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata 4501 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg 4561 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac 4621 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc 4681 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat 4741 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccgggc tcgatcccct 4801 cgcgagttgg ttcagctgct gcctgaggct ggacgacctc gcggagttct accggcagtg 4861 caaatccgtc ggcatccagg aaaccagcag cggctatccg cgcatccatg cccccgaact 4921 gcaggagtgg ggaggcacga tggccgcttt ggtcgacccg gacgggacgc tcctgcgcct 4981 gatacagaac gaattgcttg caggcatctc atgagtgtgt cttcccgttt tccgcctgag 5041 gtcactgcgt ggatggagcg ctggcgcctg ctgcgcgacg gcgagctgct caccacccac 5101 tcgccaagct ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgccga 5161 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc 5221 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag 5281 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 5341 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca 5401 attccctata gtgagtcgta ttaaattcgt aatcatgtca tagctgtttc ctgtgtgaaa 5461 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 5521 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 5581 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 5641 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 5701 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 5761 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 5821 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 5881 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 5941 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 6001 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 6061 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 6121 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 6181 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 6241 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 6301 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 6361 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 6421 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 6481 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 6541 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 6601 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 6661 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 6721 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 6781 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 6841 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 6901 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 6961 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 7021 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 7081 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 7141 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 7201 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 7261 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 7321 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 7381 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 7441 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 7501 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 7561 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc 7621 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc 7681 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa 7741 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact 7801 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt 7861 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa 7921 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt 7981 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac 8041 aattt //