LOCUS Exported 6594 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA-Wnt8B SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6594) 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 6594) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6594 /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(2034..2058) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2084..2125 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(2165..2184) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 2209..2257 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 2459..2887 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2546..2565) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2756..2777 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2896..2916) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2901..3230 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3081..3216 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3143..3162 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3297..4091 /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(3351..3370) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3961..3980 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4267..4388 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4304..4323) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4358..4377 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4437..4453) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4437..4453) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4450..4472) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4461..4477 /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(4485..4515) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4530..4551 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4668..4685) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4839..5427) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4919..4938) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5598..6458) /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 6221..6240 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6459..6563) /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 tcaccatgtt 961 tctttcaaag ccttctgtgt acatctgtct tttcacctgt gtcctccaac tcagccacag 1021 ctggtcggtg aacaatttcc tgatgactgg tccaaaggct tacctgattt actccagcag 1081 tgtggcagct ggtgcccaga gtggtattga agaatgcaag tatcagtttg cctgggaccg 1141 ctggaactgc cctgagagag ccctgcagct gtccagccat ggtgggcttc gcagtgccaa 1201 tcgggagaca gcatttgtgc atgccatcag ttctgctgga gtcatgtaca ccctgactag 1261 aaactgcagc cttggagatt ttgataactg tggctgtgat gactcccgca acgggcaact 1321 ggggggacaa ggctggctgt ggggaggctg cagtgacaat gtgggcttcg gagaggcgat 1381 ttccaagcag tttgtcgatg ccctggaaac aggacaggat gcacgggcag ccatgaacct 1441 gcacaacaac gaggctggcc gcaaggcggt gaagggcacc atgaaacgca cgtgcaagtg 1501 ccacggcgtg tctggcagct gcaccacgca gacctgttgg ctgcagctgc ccgagttccg 1561 cgaggtgggc gcgcacctga aggagaagta ccacgcagca ctcaaggtgg acctgctgca 1621 gggtgctggc aacagcgcgg ccggccgcgg cgccatcgcc gacacctttc gctccatctc 1681 tacccgggag ctggtgcacc tggaggactc cccggactac tgcctggaga acaaaacgct 1741 agggctgctg ggcaccgaag gccgagagtg cctaaggcgc gggcgggccc tgggtcgctg 1801 ggaacgccgc agctgccgcc ggctctgcgg ggactgcggg ctggcggtgg aggagcgccg 1861 ggccgagacc gtgtccagct gcaactgcaa gttccactgg tgctgcgcag tccgctgcga 1921 gcagtgccgc cggagggtca ccaagtactt ctgtagccgc gcagagcggc cgcggggggg 1981 cgctgcgcac aaacccggga gaaaaccctg aattctgaag ggtgggcgcg ccgacccagc 2041 tttcttgtac aaagtggttg atctagaggg cccgcggttc gaaggtaagc ctatccctaa 2101 ccctctcctc ggtctcgatt ctacgcgtac cggttagtaa tgagtttaaa cgggggaggc 2161 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa 2221 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag 2281 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct 2341 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc 2401 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac 2461 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct 2521 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg 2581 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt 2641 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca 2701 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga 2761 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa 2821 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac 2881 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag 2941 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 3001 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 3061 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 3121 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc 3181 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg 3241 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga 3301 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct 3361 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc 3421 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg 3481 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg 3541 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc 3601 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc 3661 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg 3721 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc 3781 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg 3841 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc 3901 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag 3961 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg 4021 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg 4081 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg 4141 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt 4201 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc 4261 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 4321 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 4381 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca 4441 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga 4501 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg 4561 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 4621 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 4681 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 4741 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 4801 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 4861 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 4921 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 4981 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca 5041 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 5101 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 5161 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 5221 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg 5281 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 5341 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 5401 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 5461 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 5521 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 5581 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 5641 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 5701 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 5761 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 5821 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 5881 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 5941 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 6001 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 6061 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 6121 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 6181 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 6241 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 6301 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 6361 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 6421 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 6481 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 6541 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc //