LOCUS V033892 7446 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033892 VERSION V033892 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" promoter 678..696 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 756..2126 /label="Foxn3(NM_183186)" /note="Foxn3(NM_183186)" /gene="Foxn3" CDS 2133..2198 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2222..2810 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2876..2924 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3126..3554 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3568..3897 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3964..4758 /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 4934..5055 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5104..5120) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5128..5144 /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(5152..5182) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5197..5218 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5506..6094) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6265..7125) /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(7126..7230) /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 gctctctggc taactagaga acccactgct 661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta 721 aacttaagct tggtaccgag ctcgaattcg ccaccatggg tccagtcatg cctgccagta 781 agaaggccga gagctcagga atcagtgtgt ccagtggact gagtcagcgt taccggggca 841 gcggtttctc caaggccctc caggaagacg atgatcttga cttccctctg cccgacatcc 901 gattagaaga gggggccatg gaagatgaag agctgaccaa cctgaactgg ctgcatgaga 961 gcaagaactt gctgaagagc tttggggagt cggtccttag gagtgtcagt cctgtgcagg 1021 acctagacga tgacacccca ccatcccctg cccactcgga catgccctat gatgccaggc 1081 agaaccccaa ctgcaagccc ccctactcct ttagctgcct catatttatg gccatcgagg 1141 actctccgac caagcgcctt ccagtgaagg acatctacaa ctggatctta gaacatttcc 1201 cgtattttgc aaatgcacct actgggtgga agaactcagt gagacacaac ttgtcattga 1261 ataagtgttt caagaaagtg gacaaagaga ggagtcagag tattgggaaa gggtcattgt 1321 ggtgcatcga cccagagtat agacaaaatc taattcaggc tctgaaaaag acaccttacc 1381 acccacctcc tactcctcag gcttatcaaa gcacatcagg tccacccatc tggccgggca 1441 gtaccttctt caagagaaac ggagccctcc tgcaagtttc tccaggagtg atacaaaatg 1501 gagcgagggt tctgagtcga gggctgtttc ctggtgtccg gccgttacca atcactccca 1561 ttgggatgac ggcagccata aggaatagca tcaccagttg caggatgcgg actgagagcg 1621 agcccccatg tggctcccca gtggtcagtg gcgatccgaa ggaggaccac aactacagta 1681 gtgccaagtc ctccactgcc cggagcacct ctcccaccag cgactccatc tcctcttcct 1741 cctcctcagc cgacgaccac tatgagtttg ctacgaaggg gagccaggag ggcagtgagg 1801 gaagcttcca gagccatgag agccacagtg aaccagagga ggaggaccgg aaacccagcc 1861 cgaaggaagg caaggatgcc ctgggggaca gcggatacgc atcccagcac aagaagcgcc 1921 agcacttcgc caaggccagg aaggtcccca gcgacacact gcccctcaaa aagagacgca 1981 cggagaagcc gcccgagagc gatgacgagg agatgaagga ggcggccggt tcactcctgc 2041 acttagcagg gattcggtcc tgtttgaaca acatcaccaa tcggacggcc aaggggcaga 2101 aagaacaaaa ggaaaccgca aaaaatggat ccgactacaa agaccatgac ggtgattata 2161 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga 2221 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 2281 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 2341 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 2401 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 2461 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 2521 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 2581 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct 2641 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 2701 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 2761 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg 2821 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca 2881 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg 2941 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg 3001 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc 3061 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta 3121 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 3181 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 3241 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg 3301 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 3361 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 3421 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 3481 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 3541 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 3601 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 3661 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 3721 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 3781 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 3841 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 3901 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt 3961 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 4021 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 4081 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 4141 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 4201 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 4261 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 4321 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 4381 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 4441 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 4501 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 4561 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 4621 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 4681 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 4741 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc 4801 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg 4861 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt 4921 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat 4981 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 5041 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc 5101 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 5161 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 5221 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 5281 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 5341 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 5401 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 5461 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 5521 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 5581 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 5641 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 5701 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 5761 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 5821 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 5881 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 5941 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 6001 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 6061 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 6121 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 6181 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 6241 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 6301 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 6361 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 6421 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 6481 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 6541 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 6601 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 6661 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 6721 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 6781 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 6841 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 6901 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 6961 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 7021 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 7081 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 7141 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 7201 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 7261 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat 7321 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc 7381 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg 7441 cttagg //