LOCUS Exported 6681 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA-Wnt5A-V5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6681) 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 6681) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6681 /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(2121..2145) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2171..2212 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(2252..2271) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 2296..2344 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 2546..2974 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2633..2652) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2843..2864 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2983..3003) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2988..3317 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3168..3303 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3230..3249 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3384..4178 /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(3438..3457) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4048..4067 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4354..4475 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4391..4410) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4445..4464 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4524..4540) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4524..4540) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4537..4559) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4548..4564 /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(4572..4602) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4617..4638 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4755..4772) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4926..5514) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5006..5025) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5685..6545) /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 6308..6327 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6546..6650) /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 tcaccatgaa 961 gaagtccatt ggaatattaa gcccaggagt tgctttgggg atggctggaa gtgcaatgtc 1021 ttccaagttc ttcctagtgg ctttggccat atttttctcc ttcgcccagg ttgtaattga 1081 agccaattct tggtggtcgc taggtatgaa taaccctgtt cagatgtcag aagtatatat 1141 tataggagca cagcctctct gcagccaact ggcaggactt tctcaaggac agaagaaact 1201 gtgccacttg tatcaggacc acatgcagta catcggagaa ggcgcgaaga caggcatcaa 1261 agaatgccag tatcaattcc gacatcgaag gtggaactgc agcactgtgg ataacacctc 1321 tgtttttggc agggtgatgc agataggcag ccgcgagacg gccttcacat acgcggtgag 1381 cgcagcaggg gtggtgaacg ccatgagccg ggcgtgccgc gagggcgagc tgtccacctg 1441 cggctgcagc cgcgccgcgc gccccaagga cctgccgcgg gactggctct ggggcggctg 1501 cggcgacaac atcgactatg gctaccgctt tgccaaggag ttcgtggacg cccgcgagcg 1561 ggagcgcatc cacgccaagg gctcctacga gagtgctcgc atcctcatga acctgcacaa 1621 caacgaggcc ggccgcagga cggtgtacaa cctggctgat gtggcctgca agtgccatgg 1681 ggtgtccggc tcatgtagcc tgaagacatg ctggctgcag ctggcagact tccgcaaggt 1741 gggtgatgcc ctgaaggaga agtacgacag cgcggcggcc atgcggctca acagccgggg 1801 caagttggta caggtcaaca gccgcttcaa ctcgcccacc acacaagacc tggtctacat 1861 cgaccccagc cctgactact gcgtgcgcaa tgagagcacc ggctcgctgg gcacgcaggg 1921 ccgcctgtgc aacaagacgt cggagggcat ggatggctgc gagctcatgt gctgcggccg 1981 tggctacgac cagttcaaga ccgtgcagac ggagcgctgc cactgcaagt tccactggtg 2041 ctgctacgtc aagtgcaaga agtgcacgga gatcgtggac cagtttgtgt gcaagtgcat 2101 tctgaagggt gggcgcgccg acccagcttt cttgtacaaa gtggttgatc tagagggccc 2161 gcggttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg 2221 ttagtaatga gtttaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag 2281 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt 2341 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc 2401 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt 2461 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc 2521 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt 2581 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 2641 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 2701 ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 2761 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 2821 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 2881 atctcggtct attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa 2941 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag 3001 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 3061 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 3121 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 3181 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 3241 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag 3301 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag 3361 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg 3421 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg 3481 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc 3541 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga 3601 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc 3661 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag 3721 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat 3781 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg 3841 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca 3901 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct 3961 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg 4021 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg 4081 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc 4141 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa 4201 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc 4261 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc 4321 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt 4381 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 4441 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct 4501 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 4561 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 4621 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 4681 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 4741 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 4801 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 4861 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 4921 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 4981 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 5041 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 5101 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 5161 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 5221 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 5281 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 5341 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca 5401 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 5461 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 5521 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 5581 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 5641 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 5701 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 5761 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 5821 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 5881 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 5941 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 6001 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 6061 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 6121 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 6181 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 6241 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 6301 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 6361 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 6421 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 6481 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 6541 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 6601 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 6661 cgaaaagtgc cacctgacgt c //