LOCUS Exported 6738 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS tau/pET29b SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6738) AUTHORS Hedgepeth CM, Conrad LJ, Zhang J, Huang HC, Lee VM, Klein PS TITLE Activation of the Wnt signaling pathway: a molecular mechanism for lithium action. JOURNAL Dev Biol. 1997 May 1. 185(1):82-91. PUBMED 9169052 REFERENCE 2 (bases 1 to 6738) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6738 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 563..582 /label=LacI-R /note="LacI, reverse primer" promoter complement(602..679) /gene="lacI" /label=lacI promoter primer_bind 885..904 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 992..1011 /label=T7 /note="T7 promoter, forward primer" promoter 992..1010 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 1011..1035 /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)." RBS 1050..1072 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 2590..2607 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(2660..2678) /label=T7 Term /note="T7 terminator, reverse primer" terminator 2674..2721 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 2758..3213 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2845..2864) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3055..3076 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(3305..4120) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 4028..4047 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 4112..4131 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 4242..4830 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4731..4750 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4984..5001 /label=L4440 /note="L4440 vector, forward primer" misc_feature 5016..5155 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(5081..5100) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5241..5263 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(5257..5448) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(6257..601) /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" ORIGIN 1 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt 61 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga 121 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag 181 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc 241 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc 301 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg 361 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg 421 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg 481 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg 541 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca 601 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt 661 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg 721 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc 781 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa 841 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata 901 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag 961 aggatcgaga tcgatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga 1021 gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga gatatacata 1081 tggctgagcc ccgccaggag ttcgaagtga tggaagatca cgctgggacg tacgggttgg 1141 gggacaggaa agatcagggg ggctacacca tgcaccaaga ccaagagggt gacacggacg 1201 ctggcctgaa agaatctccc ctgcagaccc ccactgagga cggatctgag gaaccgggct 1261 ctgaaacctc tgatgctaag agcactccaa cagcggaaga tgtgacagca cccttagtgg 1321 atgagggagc tcccggcaag caggctgccg cgcagcccca cacggagatc ccagaaggaa 1381 ccacagctga agaagcaggc attggagaca cccccagcct ggaagacgaa gctgctggtc 1441 acgtgaccca agctcgcatg gtcagtaaaa gcaaagacgg gactggaagc gatgacaaaa 1501 aagccaaggg ggctgatggt aaaacgaaga tcgccacacc gcggggagca gcccctccag 1561 gccagaaggg ccaggccaac gccaccagga ttccagcaaa aaccccgccc gctccaaaga 1621 caccacccag ctctggtgaa cctccaaaat caggggatcg cagcggctac agcagccccg 1681 gctccccagg cactcccggc agccgctccc gcaccccgtc ccttccaacc ccacccaccc 1741 gggagcccaa gaaggtggca gtggtccgta ctccacccaa gtcgccgtct tccgccaaga 1801 gccgcctgca gacagccccc gtgcccatgc cagacctgaa gaatgtcaag tccaagatcg 1861 gctccactga gaacctgaag caccagccgg gaggcgggaa ggtgcagata attaataaga 1921 agctggatct tagcaacgtc cagtccaagt gtggctcaaa ggataatatc aaacacgtcc 1981 cgggaggcgg cagtgtgcaa atagtctaca aaccagttga cctgagcaag gtgacctcca 2041 agtgtggctc attaggcaac atccatcata aaccaggagg tggccaggtg gaagtaaaat 2101 ctgagaagct tgacttcaag gacagagtcc agtcgaagat tgggtccctg gacaatatca 2161 cccacgtccc tggcggagga aataaaaaga ttgaaaccca caagctgacc ttccgcgaga 2221 acgccaaagc caagacagac cacggggcgg agatcgtgta caagtcgcca gtggtgtctg 2281 gggacacgtc tccacggcat ctcagcaatg tctcctccac cggcagcatc gacatggtag 2341 actcgcccca gctcgccacg ctagctgacg aggtgtctgc ctccctggcc aagcagggtt 2401 tgtgatcagg cccctggggc ggtcaataat tgtggagagg agagaatgag agagtgtgga 2461 aaaaaaaaga ataatgaccc ggcccccgcc ctctgccccc agctgctcct cgcagttcgg 2521 gaattagcta agaaggagat ataggatccg aattcgagct ccgtcgacaa gcttgcggcc 2581 gcactcgagc accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa 2641 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa 2701 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg 2761 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta 2821 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt 2881 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg 2941 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat 3001 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac 3061 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag 3121 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg 3181 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg 3241 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta 3301 attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat 3361 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt 3421 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac 3481 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga 3541 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag 3601 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg 3661 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa 3721 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag 3781 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg 3841 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc 3901 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca 3961 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc 4021 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc 4081 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt 4141 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc 4201 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 4261 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 4321 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 4381 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 4441 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 4501 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 4561 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 4621 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 4681 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 4741 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 4801 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 4861 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg 4921 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag 4981 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg 5041 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg 5101 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg 5161 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta 5221 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc 5281 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat 5341 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct 5401 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt 5461 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac 5521 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact 5581 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt 5641 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa 5701 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa 5761 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg 5821 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt 5881 cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc 5941 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg 6001 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc 6061 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa 6121 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg 6181 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac 6241 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 6301 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt 6361 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga 6421 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg 6481 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatgt 6541 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat 6601 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt 6661 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc 6721 gagtgagata tttatgcc //