LOCUS V044890 8346 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044890 VERSION V044890 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..3026 /label="Dact2(NM_172826)" /note="Dact2(NM_172826)" /gene="Dact2" CDS 3033..3098 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 3122..3710 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3776..3824 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 4026..4454 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4468..4797 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4864..5658 /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 5834..5955 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6004..6020) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6028..6044 /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(6052..6082) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6097..6118 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6406..6994) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7165..8025) /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(8026..8130) /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 ccaccatgtg ggcaccgagc ggccaggggc 781 ccgcgggttg ggaccgccgc agggtgggcg cgcgactacg agccgcactg gccgggctgc 841 aggagcttca ggggctgcga gccacgcagc aagcacgcgt gcggggagct ctgggcctgc 901 accctgcgcc agggccccgc ggccaggagc tgcgtctgga ggcggcgctg accgcgctac 961 gggagcagct gtcacggcta aggagacagg atgctggcct gaagacacac ttggaccagc 1021 tggatcagca gataagtgaa ctacagctgg atgtgagcag gtcttcttgc gaggccttgg 1081 atagtgacag cagacccagc tcaggtttct atgagctgag cgatgctggt tcctgttctc 1141 tgtccacctc ctgcgcatct gtttgcagcg accgtctatc cccctccctg ggtagctggc 1201 tgcctgtgtt ccagccctcc aagtccaggt ctggcatcgg ggactggcga ccacgctctg 1261 ctgatgagac cactgtccct gcatggagcc cacagctcac agaagatagc aggctcctgc 1321 atggtgcaga gggcacaggc cggctgacgg gcatgttccg gcccagacca gtatctacag 1381 gtgatctcga aagagtcctt ccagctgacg tggggctcca gagagctggt actgatgctg 1441 cacatctcct gggccagggg atagagatcc cagcccacgc cctggacccc acgtaccagc 1501 gtgacctggt ggccagggga ggccaggagg tgtacccata tcccagcccc ctccatgcgg 1561 tggccctgca gagtcccctc ttcgctctgc ccaaagaagc cccgtgtttt gacatctgct 1621 cacctcccca ggagcctcct ctggtccctg ttgatgagaa caggactcaa cctgagccga 1681 tccgtgagct gggctcagcc gaagcctaca tccacaggct gttgcatctg cggggccaag 1741 agctccccct gagagatgtg gggcaggagc agggaggtga cacagctgct tttccaccga 1801 agccctgtgg ccagaggtcg gagagcacat gtcagctgga gaagcaggcc tgtggagctg 1861 acaggggagg actgaaacta ggtaggggtg ctgccaagga cagcctcaag caacatgggc 1921 ctgtgtccct tgtgggtgct gagcccctca gcagccccct aaaggaagaa accattcctt 1981 ggaatccctg tgtccatgga gacaatactg ttggttcctc accctgctcc caggcccaac 2041 agcctcttaa tgactgtggc caaggaccag tcctgtcacc atccagggtg ttgggcactg 2101 agagcccacc tctggccccg gagccctttg cctatacatc ctgcaccact ggtgaaacct 2161 ctcccgtgaa gctgaggatg ggcttttccc aaaacaaggc cgtgaaggtc agaagaagag 2221 ttagtgagaa agtcccgagg ctagggaagc agctccctcc acaaccagag aggcagcggg 2281 tcacagagcg ggacccctcc aggccccatc agggaggtct cagcaggagg cccacactgg 2341 cccgggagcc tcctggacgc tcctgctctg agtccaccct ctaccccgtg cccttccttg 2401 tccccgtagt ggtggcccag agggaaagtt acccaacgtc accccaagcg ttcttcccaa 2461 tggaggcagc tctcctcagc tcagcagcca ggcggaagca gcgcaggtgg cagtccacca 2521 tggagatctc agccaaagcc ggttcagtca gccaacctgg gcccagcatg gggctcccta 2581 ggtccccagc caagagagga agtggtccca gggcccagag taggcccaca cttgcccgtc 2641 aggatgcctg tgcgaggtgt gagtcggacc cctcggagca ctctgcagac tgcacctcac 2701 tctaccactc caccattgcc gagaccagcg aggacgagga ggcgagtgac cacactgcca 2761 accgctttgg ggacgagtcc agcagcaacg attcagaagg gtgtttccgg ggcagccgcc 2821 gtcgcctggc aataggcagt gcagaggctg ggcaaggggg gtgggcctgg cctcgggtgc 2881 ccccccagca gccctcacgg gccccaggaa acaccaggcc acccttgccc cctgtgccca 2941 aactgtgccg catcaaggcc tccaaggccc tgaagaagaa gatccgaagg tttcagccag 3001 cagccctgaa ggtcatgacc atggtgggat ccgactacaa agaccatgac ggtgattata 3061 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga 3121 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 3181 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 3241 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 3301 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 3361 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 3421 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 3481 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct 3541 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 3601 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 3661 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg 3721 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca 3781 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg 3841 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg 3901 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc 3961 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta 4021 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 4081 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 4141 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg 4201 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 4261 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 4321 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 4381 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 4441 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 4501 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 4561 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 4621 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 4681 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 4741 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 4801 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt 4861 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 4921 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 4981 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 5041 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 5101 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 5161 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 5221 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 5281 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 5341 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 5401 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 5461 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 5521 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 5581 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 5641 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc 5701 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg 5761 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt 5821 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat 5881 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 5941 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc 6001 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 6061 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 6121 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 6181 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 6241 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 6301 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 6361 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 6421 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 6481 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 6541 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 6601 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 6661 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 6721 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 6781 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 6841 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 6901 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 6961 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 7021 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 7081 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 7141 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 7201 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 7261 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 7321 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 7381 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 7441 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 7501 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 7561 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 7621 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 7681 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 7741 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 7801 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 7861 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 7921 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 7981 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 8041 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 8101 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 8161 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat 8221 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc 8281 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg 8341 cttagg //