LOCUS V019707 7424 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019707 VERSION V019707 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..2104 /label="SOX4(NM_003107)" /note="SOX4(NM_003107)" /gene="SOX4" CDS 2111..2176 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2200..2788 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2854..2902 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3104..3532 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3546..3875 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3942..4736 /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 4912..5033 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5082..5098) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5106..5122 /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(5130..5160) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5175..5196 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5484..6072) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6243..7103) /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(7104..7208) /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 ccatggtgca gcaaaccaac aatgccgaga acacggaagc 721 gctgctggcc ggcgagagct cggactcggg cgccggcctc gagctgggaa tcgcctcctc 781 ccccacgccc ggctccaccg cctccacggg cggcaaggcc gacgacccga gctggtgcaa 841 gaccccgagt gggcacatca agcgacccat gaacgccttc atggtgtggt cgcagatcga 901 gcggcgcaag atcatggagc agtcgcccga catgcacaac gccgagatct ccaagcggct 961 gggcaaacgc tggaagctgc tcaaagacag cgacaagatc cctttcattc gagaggcgga 1021 gcggctgcgc ctcaagcaca tggctgacta ccccgactac aagtaccggc ccaggaagaa 1081 ggtgaagtcc ggcaacgcca actccagctc ctcggccgcc gcctcctcca agccggggga 1141 gaagggagac aaggtcggtg gcagtggcgg gggcggccat gggggcggcg gcggcggcgg 1201 gagcagcaac gcggggggag gaggcggcgg tgcgagtggc ggcggcgcca actccaaacc 1261 ggcgcagaaa aagagctgcg gctccaaagt ggcgggcggc gcgggcggtg gggttagcaa 1321 accgcacgcc aagctcatcc tggcaggcgg cggcggcggc gggaaagcag cggctgccgc 1381 cgccgcctcc ttcgccgccg aacaggcggg ggccgccgcc ctgctgcccc tgggcgccgc 1441 cgccgaccac cactcgctgt acaaggcgcg gactcccagc gcctcggcct ccgcctcctc 1501 ggcagcctcg gcctccgcag cgctcgcggc cccgggcaag cacctggcgg agaagaaggt 1561 gaagcgcgtc tacctgttcg gcggcctggg cacgtcgtcg tcgcccgtgg gcggcgtggg 1621 cgcgggagcc gaccccagcg accccctggg cctgtacgag gaggagggcg cgggctgctc 1681 gcccgacgcg cccagcctga gcggccgcag cagcgccgcc tcgtcccccg ccgccggccg 1741 ctcgcccgcc gaccaccgcg gctacgccag cctgcgcgcc gcctcgcccg ccccgtccag 1801 cgcgccctcg cacgcgtcct cctcggcctc gtcccactcc tcctcttcct cctcctcggg 1861 ctcctcgtcc tccgacgacg agttcgaaga cgacctgctc gacctgaacc ccagctcaaa 1921 ctttgagagc atgtccctgg gcagcttcag ttcgtcgtcg gcgctcgacc gggacctgga 1981 ttttaacttc gagcccggct ccggctcgca cttcgagttc ccggactact gcacgcccga 2041 ggtgagcgag atgatctcgg gagactggct cgagtccagc atctccaacc tggttttcac 2101 ctacctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa 2161 ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct ggattacaaa 2221 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 2281 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 2341 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 2401 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 2461 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 2521 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 2581 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 2641 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 2701 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt 2761 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga 2821 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc 2881 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc 2941 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc 3001 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct 3061 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc 3121 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 3181 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 3241 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc 3301 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 3361 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 3421 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt 3481 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt 3541 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 3601 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 3661 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 3721 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 3781 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt 3841 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca 3901 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat 3961 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 4021 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 4081 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc 4141 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 4201 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 4261 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 4321 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 4381 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc 4441 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga 4501 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 4561 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 4621 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 4681 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 4741 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc 4801 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc 4861 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt 4921 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 4981 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc 5041 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 5101 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 5161 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 5221 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 5281 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 5341 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 5401 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 5461 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 5521 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 5581 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 5641 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc 5701 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 5761 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 5821 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 5881 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc 5941 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 6001 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 6061 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 6121 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 6181 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 6241 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 6301 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 6361 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 6421 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 6481 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 6541 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 6601 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 6661 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 6721 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 6781 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 6841 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 6901 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 6961 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 7021 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 7081 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 7141 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 7201 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg 7261 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc 7321 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa 7381 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg //