LOCUS Exported 6636 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA-Wnt6-V5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6636) 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 6636) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6636 /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(2076..2100) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2126..2167 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(2207..2226) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 2251..2299 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 2501..2929 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2588..2607) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2798..2819 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2938..2958) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2943..3272 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3123..3258 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3185..3204 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3339..4133 /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(3393..3412) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4003..4022 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4309..4430 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4346..4365) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4400..4419 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4479..4495) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4479..4495) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4492..4514) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4503..4519 /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(4527..4557) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4572..4593 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4710..4727) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4881..5469) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4961..4980) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5640..6500) /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 6263..6282 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6501..6605) /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 gccgccctta ccctcccgcc tcgggctgct gctgctgctg ctcctgtgcc cggcgcacgt 1021 cggcggactg tggtgggctg tgggcagccc cttggttatg gaccctacca gcatctgcag 1081 gaaggcacgg cggctggccg ggcggcaggc cgagttgtgc caggctgagc cggaagtggt 1141 ggcagagcta gctcggggcg cccggctcgg ggtgcgagag tgccagttcc agttccgctt 1201 ccgccgctgg aattgctcca gccacagcaa ggcctttgga cgcatcctgc aacaggacat 1261 tcgggagacg gccttcgtgt tcgccatcac tgcggccggc gccagccacg ccgtcacgca 1321 ggcctgttct atgggcgagc tgctgcagtg cggctgccag gcgccccgcg ggcgggcccc 1381 tccccggccc tccggcctgc ccggcacccc cggaccccct ggccccgcgg gctccccgga 1441 aggcagcgcc gcctgggagt ggggaggctg cggcgacgac gtggacttcg gggacgagaa 1501 gtcgaggctc tttatggacg cacggcacaa gcggggacgc ggagacatcc gcgcgttggt 1561 gcaactgcac aacaacgagg cgggcaggct ggccgtgcgg agccacacgc gcaccgagtg 1621 caaatgccac gggctgtcgg gatcatgcgc gctgcgcacc tgctggcaga agctgcctcc 1681 atttcgcgag gtgggcgcgc ggctgctgga gcgcttccac ggcgcctcac gcgtcatggg 1741 caccaacgac ggcaaggccc tgctgcccgc cgtccgcacg ctcaagccgc cgggccgagc 1801 ggacctcctc tacgccgccg attcgcccga cttctgcgcc cccaaccgac gcaccggctc 1861 ccccggcacg cgcggtcgcg cctgcaatag cagcgccccg gacctcagcg gctgcgacct 1921 gctgtgctgc ggccgcgggc accgccagga gagcgtgcag ctcgaagaga actgcctgtg 1981 ccgcttccac tggtgctgcg tagtacagtg ccaccgctgc cgtgtgcgca aggagctcag 2041 cctctgcctg tgcattctga agggtgggcg cgccgaccca gctttcttgt acaaagtggt 2101 tgatctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc tcggtctcga 2161 ttctacgcgt accggttagt aatgagttta aacgggggag gctaactgaa acacggaagg 2221 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg 2281 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat 2341 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac 2401 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc 2461 atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca 2521 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 2581 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 2641 caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac 2701 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 2761 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 2821 acaacactca accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg 2881 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga 2941 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3001 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 3061 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 3121 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 3181 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag 3241 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt 3301 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc 3361 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga 3421 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt 3481 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat 3541 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg 3601 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg 3661 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc 3721 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga 3781 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag 3841 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc 3901 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg 3961 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata 4021 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg 4081 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac 4141 tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat 4201 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg 4261 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt 4321 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 4381 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt 4441 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga 4501 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 4561 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 4621 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 4681 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 4741 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 4801 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 4861 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 4921 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 4981 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 5041 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt 5101 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 5161 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 5221 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 5281 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc 5341 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 5401 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 5461 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 5521 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 5581 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 5641 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 5701 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 5761 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 5821 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 5881 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 5941 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 6001 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 6061 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 6121 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 6181 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 6241 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 6301 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 6361 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 6421 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 6481 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 6541 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 6601 ccgcgcacat ttccccgaaa agtgccacct gacgtc //