LOCUS Exported 6597 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA-Wnt3A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6597) 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 6597) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6597 /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(2037..2061) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2087..2128 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(2168..2187) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 2212..2260 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 2462..2890 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2549..2568) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2759..2780 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2899..2919) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2904..3233 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3084..3219 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3146..3165 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3300..4094 /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(3354..3373) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3964..3983 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4270..4391 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4307..4326) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4361..4380 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4440..4456) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4440..4456) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4453..4475) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4464..4480 /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(4488..4518) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4533..4554 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4671..4688) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4842..5430) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4922..4941) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5601..6461) /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 6224..6243 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6462..6566) /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 gctggctagt 901 taagctatca acaagtttgt acaaaaaagc aggctccgcg gccgccccct tcaccatggc 961 cccactcgga tacttcttac tcctctgcag cctgaagcag gctctgggca gctacccgat 1021 ctggtggtcg ctggctgttg ggccacagta ttcctccctg ggctcgcagc ccatcctgtg 1081 tgccagcatc ccgggcctgg tccccaagca gctccgcttc tgcaggaact acgtggagat 1141 catgcccagc gtggccgagg gcatcaagat tggcatccag gagtgccagc accagttccg 1201 cggccgccgg tggaactgca ccaccgtcca cgacagcctg gccatcttcg ggcccgtgct 1261 ggacaaagct accagggagt cggcctttgt ccacgccatt gcctcagccg gtgtggcctt 1321 tgcagtgaca cgctcatgtg cagaaggcac ggccgccatc tgtggctgca gcagccgcca 1381 ccagggctca ccaggcaagg gctggaagtg gggtggctgt agcgaggaca tcgagtttgg 1441 tgggatggtg tctcgggagt tcgccgacgc ccgggagaac cggccagatg cccgctcagc 1501 catgaaccgc cacaacaacg aggctgggcg ccaggccatc gccagccaca tgcacctcaa 1561 gtgcaagtgc cacgggctgt cgggcagctg cgaggtgaag acatgctggt ggtcgcaacc 1621 cgacttccgc gccatcggtg acttcctcaa ggacaagtac gacagcgcct cggagatggt 1681 ggtggagaag caccgggagt cccgcggctg ggtggagacc ctgcggccgc gctacaccta 1741 cttcaaggtg cccacggagc gcgacctggt ctactacgag gcctcgccca acttctgcga 1801 gcccaaccct gagacgggct ccttcggcac gcgcgaccgc acctgcaacg tcagctcgca 1861 cggcatcgac ggctgcgacc tgctgtgctg cggccgcggc cacaacgcgc gagcggagcg 1921 gcgccgggag aagtgccgct gcgtgttcca ctggtgctgc tacgtcagct gccaggagtg 1981 cacgcgcgtc tacgacgtgc acacctgcaa gtgaattctg aagggtgggc gcgccgaccc 2041 agctttcttg tacaaagtgg ttgatctaga gggcccgcgg ttcgaaggta agcctatccc 2101 taaccctctc ctcggtctcg attctacgcg taccggttag taatgagttt aaacggggga 2161 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa 2221 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc 2281 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt 2341 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac 2401 gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc 2461 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 2521 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 2581 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt 2641 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg 2701 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 2761 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta 2821 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt 2881 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc 2941 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt 3001 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 3061 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc 3121 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg 3181 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc 3241 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca 3301 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg 3361 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag 3421 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc 3481 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc 3541 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg 3601 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc 3661 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca 3721 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag 3781 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg 3841 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg 3901 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca 3961 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc 4021 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg 4081 acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac 4141 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc 4201 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc 4261 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 4321 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 4381 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg 4441 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 4501 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 4561 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 4621 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 4681 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 4741 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 4801 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 4861 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 4921 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 4981 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 5041 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 5101 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 5161 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 5221 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 5281 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 5341 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 5401 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 5461 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 5521 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 5581 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 5641 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 5701 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 5761 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 5821 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 5881 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 5941 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 6001 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 6061 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 6121 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 6181 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 6241 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 6301 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 6361 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 6421 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 6481 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 6541 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc //