LOCUS Exported 6264 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Bacterial expression for structure determination; may not be full ORF. ACCESSION . VERSION . KEYWORDS NUDT22 (NUDT22A-c001) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6264) TITLE SGC Oxford 2017 REFERENCE 2 (bases 1 to 6264) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6264 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..20 /label=T7 /note="T7 promoter, forward primer" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 20..44 /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 59..81 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 92..109 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 134..154 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1129..1146 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1199..1217) /label=T7 Term /note="T7 terminator, reverse primer" terminator 1213..1260 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 1297..1752 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1384..1403) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1594..1615 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(1844..2659) /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 2567..2586 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 2651..2670 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 2781..3369 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3270..3289 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3523..3540 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3555..3694 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3620..3639) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3780..3802 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3796..3987) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4796..5878) /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 5840..5859 /label=LacI-R /note="LacI, reverse primer" promoter complement(5879..5956) /gene="lacI" /label=lacI promoter primer_bind 6162..6181 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt 61 tgtttaactt taagaaggag atatacatat gcaccatcat catcatcatt cttctggtgt 121 agatctgggt accgagaacc tgtacttcca atccatggat cctgaggtga ccttgctgct 181 gcagtgccct ggcgggggcc tgccccagga gcagatacag gccgagctga gccccgccca 241 tgaccgtcgc ccactgccag gtggggacga ggccatcact gccatctggg agacccggct 301 aaaggcccaa ccctggctct tcgacgcccc caagttccgc ctgcactcag ccaccctggc 361 gcctattggc tctcgggggc cacagctgct cctgcgcctg ggccttactt cctaccgaga 421 cttcctgggc accaactggt ccagctcagc tgcctggctg cgacagcagg gtgccaccga 481 ctggggtgac acgcaggcct atctggcgga cccactgggg gtgggcgctg cactagccac 541 agccgatgac ttccttgtct tcctgcgccg ctcccggcag gtggctgagg cccctgggct 601 ggtggacgta cctggtgggc accctgagcc tcaggccctg tgccctggtg gcagccccca 661 gcaccaggac ctcgctgggc agctggtggt acatgaactc ttttccagtg tccttcagga 721 gatctgtgat gaggtgaacc tgccgctgct caccctgagc cagcccctgc tgttgggcat 781 cgcccgaaat gagaccagtg ctggccgagc cagtgccgag ttctatgtcc agtgcagcct 841 gacttctgag caggtgagga agcactacct gagtggggga cccgaggccc acgagtctac 901 aggaatcttc tttgtggaga cacagaacgt gcggagattg cccgagacgg agatgtgggc 961 tgaactctgc ccctcggcca aaggcgccat catcctctac aaccgggttc agggaagtcc 1021 cactggagcg gccctagggt ccccagccct actcccgccg ctctgacagt aaaggtggat 1081 acggatccga attcgagctc cgtcgacaag cttgcggccg cactcgagca ccaccaccac 1141 caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc 1201 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg 1261 ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa 1321 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 1381 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 1441 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 1501 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 1561 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 1621 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 1681 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa 1741 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt 1801 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga 1861 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa 1921 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct 1981 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt 2041 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg 2101 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat 2161 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa 2221 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga 2281 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga 2341 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa 2401 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat 2461 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg 2521 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt 2581 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt 2641 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta 2701 ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta 2761 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa 2821 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt 2881 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag 2941 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta 3001 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca 3061 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag 3121 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa 3181 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga 3241 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc 3301 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc 3361 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt 3421 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt 3481 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag 3541 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac 3601 cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac 3661 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga 3721 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc 3781 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg 3841 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc 3901 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt 3961 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat 4021 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga 4081 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga 4141 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc 4201 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga 4261 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca 4321 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt 4381 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat 4441 catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt 4501 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag 4561 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag 4621 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac 4681 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat 4741 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact 4801 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 4861 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg 4921 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc 4981 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg 5041 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg 5101 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag 5161 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc 5221 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc 5281 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct 5341 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa 5401 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc 5461 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac 5521 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt 5581 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga 5641 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact 5701 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg 5761 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa 5821 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat 5881 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc 5941 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag 6001 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag 6061 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa 6121 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag 6181 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg 6241 atcgagatct cgatcccgcg aaat //