LOCUS Exported 6792 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA-Wnt10A-V5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6792) 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 6792) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6792 /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(2232..2256) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2282..2323 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(2363..2382) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 2407..2455 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 2657..3085 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2744..2763) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2954..2975 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3094..3114) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3099..3428 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3279..3414 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3341..3360 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3495..4289 /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(3549..3568) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4159..4178 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4465..4586 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4502..4521) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4556..4575 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4635..4651) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4635..4651) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4648..4670) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4659..4675 /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(4683..4713) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4728..4749 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4866..4883) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5037..5625) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5117..5136) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5796..6656) /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 6419..6438 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6657..6761) /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 tcaccatggg 961 cagcgcccac cctcgcccct ggctgcggct ccgaccccag ccccagccgc ggccagcgct 1021 ctgggtgctc ctgttcttcc tactgctgct ggctgctgcc atgcccaggt cagcacccaa 1081 tgacattctg gacctccgcc tccccccgga gcccgtgctc aatgccaaca cagtgtgcct 1141 aacattgcca ggcctgagcc ggcggcagat ggaggtgtgt gtgcgtcacc ctgatgtggc 1201 tgcctcagcc atacagggca tccagatcgc catccacgaa tgccaacacc aattcaggga 1261 ccagcgctgg aactgctcaa gcctggagac tcgcaacaag atcccctatg agagtcccat 1321 cttcagcaga ggtttccgag agagcgcttt tgcctacgcc atcgcagcag ctggcgtggt 1381 gcacgccgtg tccaatgcgt gtgccctggg caaactgaag gcctgtggct gtgatgcgtc 1441 ccggcgaggg gacgaggagg ccttccgtag gaagctgcac cgcttacaac tggatgcact 1501 gcagcgtggt aagggcctga gccatggggt cccggaacac ccagccctgc ccacagccag 1561 cccaggcctg caggactcct gggagtgggg cggctgcagc cccgacatgg gcttcgggga 1621 gcgcttttct aaggactttc tggactcccg ggagcctcac agagacatcc acgcgagaat 1681 gaggcttcac aacaaccgag ttgggaggca ggcagtgatg gagaacatgc ggcggaagtg 1741 caagtgccac ggcacgtcag gcagctgcca gctcaagacg tgctggcagg tgacgcccga 1801 gttccgcacc gtgggggcgc tgctgcgcag ccgcttccac cgcgccacgc tcatccggcc 1861 gcacaaccgc aacggcggcc agctggagcc gggcccagcg ggggcaccct cgccggctcc 1921 gggcgctccc gggccgcgcc gacgggccag ccccgccgac ctggtctact tcgaaaagtc 1981 tcccgacttc tgcgagcgcg agccgcgcct ggactcggcg ggcaccgtgg gccgcctgtg 2041 caacaagagc agcgccggct cggatggctg cggcagcatg tgctgcggcc gcggccacaa 2101 catcctgcgc cagacgcgca gcgagcgctg ccactgccgc ttccactggt gctgtttcgt 2161 ggtctgcgaa gagtgccgca tcaccgagtg ggtcagcgtc tgcaagtgca ttctgaaggg 2221 tgggcgcgcc gacccagctt tcttgtacaa agtggttgat ctagagggcc cgcggttcga 2281 aggtaagcct atccctaacc ctctcctcgg tctcgattct acgcgtaccg gttagtaatg 2341 agtttaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg 2401 ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa 2461 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc 2521 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca 2581 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc 2641 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 2701 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 2761 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct 2821 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 2881 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 2941 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc 3001 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg 3061 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa 3121 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 3181 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3241 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 3301 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 3361 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 3421 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat 3481 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 3541 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg 3601 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg 3661 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc 3721 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg 3781 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca 3841 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc 3901 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg 3961 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg 4021 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata 4081 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg 4141 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat 4201 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct 4261 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac 4321 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg 4381 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc 4441 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa 4501 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 4561 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc 4621 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 4681 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 4741 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 4801 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 4861 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 4921 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 4981 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 5041 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 5101 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 5161 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 5221 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 5281 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 5341 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 5401 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 5461 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc 5521 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 5581 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 5641 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 5701 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 5761 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 5821 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 5881 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 5941 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 6001 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 6061 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 6121 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 6181 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 6241 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 6301 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 6361 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 6421 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 6481 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 6541 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 6601 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 6661 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 6721 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 6781 ccacctgacg tc //