LOCUS Exported 6138 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS BARD1 26-327 pET28a SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6138) AUTHORS Brzovic PS, Lissounov A, Christensen DE, Hoyt DW, Klevit RE TITLE A UbcH5/ubiquitin noncovalent complex is required for processive BRCA1-directed ubiquitination. JOURNAL Mol Cell. 2006 Mar 17. 21(6):873-80. PUBMED 16543155 REFERENCE 2 (bases 1 to 6138) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6138 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 26..73 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind 69..87 /label=T7 Term /note="T7 terminator, reverse primer" CDS complement(140..157) /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" RBS 1079..1101 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" protein_bind 1116..1140 /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)." primer_bind complement(1140..1159) /label=T7 /note="T7 promoter, forward primer" promoter complement(1141..1159) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1243..1262) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" promoter 1468..1545 /gene="lacI" /label=lacI promoter CDS 1546..2628 /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" primer_bind complement(1565..1584) /label=LacI-R /note="LacI, reverse primer" CDS 3437..3628 /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" primer_bind complement(3622..3644) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 3730..3869 /label=bom /note="basis of mobility region from pBR322" primer_bind 3785..3804 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(3884..3901) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4055..4643) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4135..4154) /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind complement(4754..4773) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 4765..5580 /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 complement(4838..4857) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin complement(5672..6127) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5809..5830) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6021..6040 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa 61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt 121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagttatcta ttgtgatggc 181 cacgttttcc attattttct aatggcaaag atttcttaga tgtaagataa tttttgcaga 241 ccttctcagg agtcactact tcattcctgc tcttagtgtc tggagactct atttgctcag 301 ccaatggtaa agagacttca gttaaacttc caaaacattc agattctgtc aaggagccac 361 ttgctagtaa gtctatttca ccatttatct gaggactgga gataacagat ggttggctac 421 agaaggatac cagcttttgc ttagattcct ctttggagtc aaattcacca tcttcttttt 481 ctgcctctaa attccatttt tggttgattt cagctaaagt tttctttttt tgcttttttc 541 cagatcttgc agaagccttt ttagccctct cagaaacatc tgcaggagga cttggggaaa 601 caaattcata tgagtcttgc tgagcacttg catctttttt tattgcaggc tgggtttgca 661 ctgaagcttt actcacaaca tatctgactt ccttacttcg agggctaaac cacattttaa 721 ttgaattctt cttgtttcct gcatcattaa acaaactttt cctaggttta tcttctttca 781 aatctgacag ctcattgtca tgtagcaaat ttcgaagctt actacaaagt tgaatcatgc 841 tgtccagttg tctatttatc ttcaagtctt gtatccaggc cggggtgtaa cacactggac 901 atccagttcc aatgcagtca cttacacaat tactacagaa gatgtgctca catcctccta 961 aacacacagg ctctctcaga atgttagtac aacgcgagca gcgcagcagc ttctccaggc 1021 ggtcgagcgc ggcgcgactg tgggcccagg caccgcgacc atccggttcc atggtatatc 1081 tccttcttaa agttaaacaa aattatttct agaggggaat tgttatccgc tcacaattcc 1141 cctatagtga gtcgtattaa tttcgcggga tcgagatctc gatcctctac gccggacgca 1201 tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca 1261 ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta 1321 tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc 1381 ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt 1441 cgcataaggg agagcgtcga gatcccggac accatcgaat ggcgcaaaac ctttcgcggt 1501 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg 1561 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac 1621 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg 1681 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc 1741 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct 1801 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa 1861 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac 1921 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg 1981 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac 2041 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta 2101 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc 2161 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt 2221 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc 2281 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt 2341 gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg 2401 ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg 2461 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa 2521 agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca 2581 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat 2641 taatgtaagt tagctcactc attaggcacc gggatctcga ccgatgccct tgagagcctt 2701 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac 2761 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg 2821 cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat 2881 cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa 2941 gcaggccatt atcgccggca tggcggcccc acgggtgcgc atgatcgtgc tcctgtcgtt 3001 gaggacccgg ctaggctggc ggggttgcct tactggttag cagaatgaat caccgatacg 3061 cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat 3121 ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat 3181 tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt 3241 attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac 3301 cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg 3361 tatcgtgagc atcctctctc gtttcatcgg tatcattacc cccatgaaca gaaatccccc 3421 ttacacggag gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat 3481 cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc 3541 agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg 3601 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg 3661 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg 3721 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac 3781 tatgcggcat cagagcagat tgtactgaga gtgcaccatt gcggtgtgaa ataccgcaca 3841 gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc 3901 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 3961 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 4021 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 4081 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 4141 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 4201 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 4261 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 4321 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 4381 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 4441 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 4501 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 4561 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 4621 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 4681 agtggaacga aaactcacgt taagggattt tggtcatgaa caataaaact gtctgcttac 4741 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc ttgctctagg 4801 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat 4861 gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg 4921 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta 4981 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat 5041 gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt attagaagaa 5101 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat 5161 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg 5221 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc 5281 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca 5341 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata 5401 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta 5461 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt 5521 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa 5581 gaattaattc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 5641 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa 5701 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa 5761 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac 5821 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag 5881 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt 5941 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg 6001 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca 6061 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag 6121 ggcgcgtccc attcgcca //