LOCUS V024121 6347 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024121 VERSION V024121 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..1837 /label="WNT2B(NM_001291880)" /note="WNT2B(NM_001291880)" /gene="WNT2B" CDS 1844..1909 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1947..2171 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2217..2645 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2659..2988 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3055..3849 /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 4023..4144 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4193..4209) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4217..4233 /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(4241..4271) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4286..4307 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4595..5180) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5351..6211) /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(6212..6316) /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 atgcgttcag tgggcgaggg 961 tgcccgagaa tggatccgag agtgtcagca ccaattccgc caccaccgct ggaactgtac 1021 caccctggac cgggaccaca ccgtctttgg ccgtgtcatg ctcagaagta gccgagaggc 1081 agcttttgta tatgccatct catcagcagg ggtagtccac gctattactc gcgcctgtag 1141 ccagggtgaa ctgagtgtgt gcagctgtga cccctacacc cgtggccgac accatgacca 1201 gcgtggggac tttgactggg gtggctgcag tgacaacatc cactacggtg tccgttttgc 1261 caaggccttc gtggatgcca aggagaagag gcttaaggat gcccgggccc tcatgaactt 1321 acataataac cgctgtggtc gcacggctgt gcggcggttt ctgaagctgg agtgtaagtg 1381 ccatggcgtg agtggttcct gtactctgcg cacctgctgg cgtgcactct cagatttccg 1441 ccgcacaggt gattacctgc ggcgacgcta tgatggggct gtgcaggtga tggccaccca 1501 agatggtgcc aacttcaccg cagcccgcca aggctatcgc cgtgccaccc ggactgatct 1561 tgtctacttt gacaactctc cagattactg tgtcttggac aaggctgcag gttccctagg 1621 cactgcaggc cgtgtctgca gcaagacatc aaaaggaaca gacggttgtg aaatcatgtg 1681 ctgtggccga gggtacgaca caactcgagt cacccgtgtt acccagtgtg agtgcaaatt 1741 ccactggtgc tgtgctgtac ggtgcaagga atgcagaaat actgtggacg tccatacttg 1801 caaagccccc aagaaggcag agtggctgga ccaaaccctc gaggactaca aagaccatga 1861 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc 1921 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg 1981 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 2041 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 2101 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt 2161 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc 2221 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 2281 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 2341 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 2401 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc 2461 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 2521 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg 2581 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 2641 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca 2701 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca 2761 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc 2821 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 2881 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta 2941 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga 3001 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt 3061 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat 3121 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag 3181 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac 3241 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac 3301 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc 3361 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg 3421 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag 3481 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat 3541 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag 3601 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc 3661 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg 3721 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg 3781 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag 3841 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat 3901 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 3961 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 4021 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 4081 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 4141 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 4201 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 4261 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 4321 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 4381 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 4441 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 4501 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 4561 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 4621 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 4681 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 4741 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 4801 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 4861 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 4921 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 4981 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 5041 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 5101 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc 5161 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 5221 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 5281 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 5341 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 5401 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 5461 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 5521 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 5581 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 5641 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 5701 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 5761 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 5821 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 5881 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 5941 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 6001 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 6061 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 6121 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 6181 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 6241 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 6301 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc //