LOCUS V022284 6398 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022284 VERSION V022284 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..1930 /label="FOXS1(NM_004118)" /note="FOXS1(NM_004118)" /gene="FOXS1" CDS 1937..1960 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" polyA_signal 1998..2222 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2268..2696 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2710..3039 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3106..3900 /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 4074..4195 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4244..4260) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4268..4284 /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(4292..4322) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4337..4358 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4646..5231) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5402..6262) /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(6263..6367) /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 atgcagcagc agcctctgcc 961 cgggcctggc gcccccacaa ctgagccaac caagcctccc tacagctaca tcgcccttat 1021 tgctatggcc atccagagct caccggggca gcgggccacc ctcagtggca tctaccgcta 1081 catcatgggc cgattcgcct tctaccgcca caaccggccc ggctggcaga acagcatccg 1141 ccacaacctg tcactcaacg agtgctttgt caaggtgccc cgcgatgacc gcaagccagg 1201 caagggcagc tactggacgc tggaccctga ctgccacgac atgtttgagc acggcagctt 1261 cctacgccgc cgccgccgct tcacccggca gacaggtgct gagggcaccc ggggccccgc 1321 caaggcacgc cgtggacccc tcagggcgac cagccaggac ccaggagtcc ccaacgccac 1381 gaccggcagg cagtgctcat tcccaccaga gctgccagat cccaagggcc taagctttgg 1441 gggtctggtg ggggccatgc cagccagtat gtgcccagca accactgatg gcaggcctcg 1501 gccacccatg gagcccaaag agatttccac gcccaagcct gcatgcccag gggagctccc 1561 cgtggccacc tcatcttcct catgcccagc gtttggcttt cctgccggct tctcagaggc 1621 tgagagtttt aataaggccc ctacgcccgt cttgtccccg gaatcaggca tcgggagcag 1681 ctaccagtgt cggctgcagg cactgaattt ttgcatgggg gctgacccag gccttgagca 1741 cctcttggcc tcagcagccc cctcccctgc accacccacc cctccaggct cactccgggc 1801 cccactgccc ctgccaactg accacaagga accctgggtt gcaggtggct tccctgtcca 1861 gggaggctcc ggctacccat tggggctgac cccctgccta taccggacgc caggaatgtt 1921 cttctttgag ctcgaggatt acaaggatga cgacgataag tagtgagggc ccgtttaaac 1981 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc 2041 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga 2101 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga 2161 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat 2221 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag 2281 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag 2341 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt 2401 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca 2461 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata 2521 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca 2581 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc 2641 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt 2701 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt 2761 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca 2821 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta 2881 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga 2941 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag 3001 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata 3061 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat 3121 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca 3181 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg 3241 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg 3301 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact 3361 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct 3421 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg 3481 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt 3541 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc 3601 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc 3661 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga 3721 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc 3781 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt 3841 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga 3901 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt 3961 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg 4021 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt 4081 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag 4141 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg 4201 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 4261 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 4321 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 4381 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 4441 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 4501 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 4561 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 4621 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 4681 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 4741 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 4801 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 4861 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 4921 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 4981 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 5041 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 5101 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 5161 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 5221 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 5281 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 5341 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 5401 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 5461 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 5521 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 5581 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 5641 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 5701 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 5761 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 5821 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 5881 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 5941 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 6001 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 6061 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 6121 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 6181 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 6241 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 6301 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 6361 ttccgcgcac atttccccga aaagtgccac ctgacgtc //