LOCUS Exported 6852 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial expression for structure determination; may not be full ORF. ACCESSION . VERSION . KEYWORDS TXNRD1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6852) TITLE SGC+year JOURNAL unpublished REFERENCE 2 (bases 1 to 6852) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6852 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 4..23 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 107..126 /label=T7 /note="T7 promoter, forward primer" promoter 107..125 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 126..150 /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 165..187 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 198..215 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 240..260 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1823..1840 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1893..1911) /label=T7 Term /note="T7 terminator, reverse primer" terminator 1907..1954 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 1991..2446 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2078..2097) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2288..2309 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(2538..3353) /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 3261..3280 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 3345..3364 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 3475..4063 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3964..3983 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4217..4234 /label=L4440 /note="L4440 vector, forward primer" misc_feature 4249..4388 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(4314..4333) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4474..4496 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(4490..4681) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(5490..6572) /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 6534..6553 /label=LacI-R /note="LacI, reverse primer" promoter complement(6573..6650) /gene="lacI" /label=lacI promoter ORIGIN 1 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc 61 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact 121 ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag 181 aaggagatat acatatgcac catcatcatc atcattcttc tggtgtagat ctgggtaccg 241 agaacctgta cttccaatcc atgaacggcc ctgaagatct tcccaagtcc tatgactatg 301 accttatcat cattggaggt ggctcaggag gtctggcagc tgctaaggag gcagcccaat 361 atggcaagaa ggtgatggtc ctggactttg tcactcccac ccctcttgga actagatggg 421 gtcttggagg aacatgtgtg aatgtgggtt gcatacctaa aaaactgatg catcaagcag 481 ctttgttagg acaagccctg caagactctc gaaattatgg atggaaagtc gaggagacag 541 ttaagcatga ttgggacaga atgatagaag ctgtacagaa tcacattggc tctttgaatt 601 ggggctaccg agtagctctg cgggagaaaa aagtcgtcta tgagaatgct tatgggcaat 661 ttattggtcc tcacaggatt aaggcaacaa ataataaagg caaagaaaaa atttattcag 721 cagagagatt tctcattgcc actggtgaaa gaccacgtta cttgggcatc cctggtgaca 781 aagaatactg catcagcagt gatgatcttt tctccttgcc ttactgcccg ggtaagaccc 841 tggttgttgg agcatcctat gtcgctttgg agtgcgctgg atttcttgct ggtattggtt 901 tagacgtcac tgttatggtt aggtccattc ttcttagagg atttgaccag gacatggcca 961 acaaaattgg tgaacacatg gaagaacatg gcatcaagtt tataagacag ttcgtaccaa 1021 ttaaagttga acaaattgaa gcagggacac caggccgact cagagtagta gctcagtcca 1081 ccaatagtga ggaaatcatt gaaggagaat ataatacggt gatgctggca ataggaagag 1141 atgcttgcac aagaaaaatt ggcttagaaa ccgtaggggt gaagataaat gaaaagactg 1201 gaaaaatacc tgtcacagat gaagaacaga ccaatgtgcc ttacatctat gccattggcg 1261 atatattgga ggataaggtg gagctcaccc cagttgcaat ccaggcagga agattgctgg 1321 ctcagaggct ctatgcaggt tccactgtca agtgtgacta tgaaaatgtt ccaaccactg 1381 tatttactcc tttggaatat ggtgcttgtg gcctttctga ggagaaagct gtggagaagt 1441 ttggggaaga aaatattgag gtttaccata gttacttttg gccattggaa tggacgattc 1501 cgtcaagaga taacaacaaa tgttatgcaa aaataatctg taatactaaa gacaatgaac 1561 gtgttgtggg ctttcacgta ctgggtccaa atgctggaga agttacacaa ggctttgcag 1621 ctgcgctcaa atgtggactg accaaaaagc agctggacag cacaattgga atccaccctg 1681 tctgtgcaga ggtattcaca acattgtctg tgaccaagcg ctctggggca agcatcctcc 1741 aggctggctg cggtggttaa cagtaaaggt ggatacggat ccgaattcga gctccgtcga 1801 caagcttgcg gccgcactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa 1861 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct 1921 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt 1981 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 2041 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 2101 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg 2161 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 2221 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 2281 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct 2341 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa 2401 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc 2461 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc 2521 cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc 2581 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac 2641 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt 2701 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa 2761 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag 2821 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg 2881 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa 2941 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt 3001 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg 3061 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata 3121 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct 3181 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc 3241 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg 3301 ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct cataacaccc 3361 cttgtattac tgtttatgta agcagacagt tttattgttc atgaccaaaa tcccttaacg 3421 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga 3481 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt 3541 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag 3601 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa 3661 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag 3721 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca 3781 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac 3841 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa 3901 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc 3961 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg 4021 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc 4081 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc 4141 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag 4201 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta 4261 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc 4321 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca 4381 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc 4441 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt 4501 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa 4561 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg 4621 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca 4681 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag 4741 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg 4801 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat 4861 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga 4921 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa 4981 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt 5041 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc 5101 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc 5161 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt 5221 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc 5281 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt 5341 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga 5401 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag 5461 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 5521 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 5581 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc 5641 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc 5701 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc 5761 actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc 5821 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt 5881 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc 5941 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca 6001 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc 6061 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca 6121 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc 6181 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc 6241 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca 6301 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc 6361 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg 6421 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca 6481 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac 6541 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc 6601 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg 6661 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt 6721 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc 6781 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc 6841 ccgatcttcc cc //