LOCUS Exported 6708 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA-Wnt10B-V5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6708) AUTHORS Najdi R, Proffitt K, Sprowl S, Kaur S, Yu J, Covey TM, Virshup DM, Waterman ML TITLE A uniform human Wnt expression library reveals a shared secretory pathway and unique signaling activities. JOURNAL Differentiation. 2012 Sep;84(2):203-13. doi: 10.1016/j.diff.2012.06.004. Epub 2012 Jul 9. PUBMED 22784633 REFERENCE 2 (bases 1 to 6708) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6708 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 911..935 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(2148..2172) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2198..2239 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(2279..2298) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 2323..2371 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 2573..3001 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2660..2679) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2870..2891 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3010..3030) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3015..3344 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3195..3330 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3257..3276 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3411..4205 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3465..3484) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4075..4094 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4381..4502 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4418..4437) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4472..4491 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4551..4567) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4551..4567) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4564..4586) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4575..4591 /label=lac operator /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-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4599..4629) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4644..4665 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4782..4799) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4953..5541) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5033..5052) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5712..6572) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6335..6354 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6573..6677) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gctggctagt 901 taagctatca acaagtttgt acaaaaaagc aggctccgcg gccgccccct tcaccatgct 961 ggaggagccc cggccgcggc ctccgccctc gggcctcgcg ggtctcctgt tcctggcgtt 1021 gtgcagtcgg gctctaagca atgagattct gggcctgaag ttgcctggcg agccgccgct 1081 gacggccaac accgtgtgct tgacgctgtc cggcctgagc aagcggcagc taggcctgtg 1141 cctgcgcaac cccgacgtga cggcgtccgc gcttcagggt ctgcacatcg cggtccacga 1201 gtgtcagcac cagctgcgcg accagcgctg gaactgctcc gcgcttgagg gcggcggccg 1261 cctgccgcac cacagcgcca tcctcaagcg cggtttccga gaaagtgctt tttccttctc 1321 catgctggct gctggggtca tgcacgcagt agccacggcc tgcagcctgg gcaagctggt 1381 gagctgtggc tgtggctgga agggcagtgg tgagcaggat cggctgaggg ccaaactgct 1441 gcagctgcag gcactgtccc gaggcaagag tttcccccac tctctgccca gccctggccc 1501 tggctcaagc cccagccctg gcccccagga cacatgggaa tggggtggct gtaaccatga 1561 catggacttt ggagagaagt tctctcggga tttcttggat tccagggaag ctccccggga 1621 catccaggca cgaatgcgaa tccacaacaa cagggtgggg cgccaggtgg taactgaaaa 1681 cctgaagcgg aaatgcaagt gtcatggcac atcaggcagc tgccagttca agacatgctg 1741 gagggcggcc ccagagttcc gggcagtggg ggcggcgttg agggagcggc tgggccgggc 1801 catcttcatt gatacccaca accgcaattc tggagccttc cagccccgtc tgcgtccccg 1861 tcgcctctca ggagagctgg tctactttga gaagtctcct gacttctgtg agcgagaccc 1921 cactatgggc tccccaggga caaggggccg ggcctgcaac aagaccagcc gcctgttgga 1981 tggctgtggc agcctgtgct gtggccgtgg gcacaacgtg ctccggcaga cacgagttga 2041 gcgctgccat tgccgcttcc actggtgctg ctatgtgctg tgtgatgagt gcaaggttac 2101 agagtgggtg aatgtgtgta agtgcattct gaagggtggg cgcgccgacc cagctttctt 2161 gtacaaagtg gttgatctag agggcccgcg gttcgaaggt aagcctatcc ctaaccctct 2221 cctcggtctc gattctacgc gtaccggtta gtaatgagtt taaacggggg aggctaactg 2281 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga 2341 ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg 2401 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt 2461 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg 2521 gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc 2581 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt 2641 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc 2701 ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta 2761 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc 2821 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg 2881 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt 2941 ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat 3001 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca 3061 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct 3121 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc 3181 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg 3241 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc 3301 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt 3361 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac 3421 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact 3481 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc 3541 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg 3601 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg 3661 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt 3721 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc 3781 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag 3841 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg 3901 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc 3961 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt 4021 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg 4081 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt 4141 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct 4201 tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca 4261 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg 4321 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc 4381 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 4441 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 4501 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg 4561 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 4621 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 4681 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 4741 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 4801 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 4861 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 4921 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 4981 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 5041 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 5101 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt 5161 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 5221 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 5281 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 5341 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta 5401 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 5461 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 5521 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 5581 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 5641 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 5701 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 5761 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 5821 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 5881 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 5941 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 6001 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 6061 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 6121 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 6181 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 6241 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 6301 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 6361 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 6421 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 6481 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 6541 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 6601 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 6661 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc //