LOCUS Exported 5853 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Bacterial expression for structure determination; may not be full ORF. ACCESSION . VERSION . KEYWORDS DCP2 (DCP2B-c016) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5853) TITLE SGC Oxford 2017 REFERENCE 2 (bases 1 to 5853) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5853 /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 718..735 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(788..806) /label=T7 Term /note="T7 terminator, reverse primer" terminator 802..849 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 886..1341 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(973..992) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1183..1204 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(1433..2248) /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 2156..2175 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 2240..2259 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 2370..2958 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2859..2878 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3112..3129 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3144..3283 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3209..3228) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3369..3391 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3385..3576) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4385..5467) /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 5429..5448 /label=LacI-R /note="LacI, reverse primer" promoter complement(5468..5545) /gene="lacI" /label=lacI promoter primer_bind 5751..5770 /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 atccatggga gtaccaacat atggtgcaat 181 tattcttgat gagacacttg aaaatgtact actagttcag gggtacctag caaaatcagg 241 ctggggattt ccaaaaggaa aagtaaataa agaagaagct cctcatgatt gtgctgctag 301 agaggtcttt gaagaaactg gttttgatat caaagactat atttgtaagg atgattacat 361 tgaacttcga atcaatgacc agcttgctcg tttgtacatc attccaggaa ttccaaaaga 421 cacaaaattt aacccaaaaa ctagaagaga aattcggaac attgagtggt tctctattga 481 gaaattgcct tgtcatagaa atgatatgac ccccaaatcc aaacttggtt tggcacctaa 541 caaatttttt atggccattc cctttatcag accattaagg gactggcttt ctcgaagatt 601 tggcgattcc tcagacagtg acaatggatt ttcctcaact ggtagcacgc cgtgacagta 661 aaggtggata cggatccgaa ttcgagctcc gtcgacaagc ttgcggccgc actcgagcac 721 caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct 781 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg 841 ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg 901 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg 961 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc 1021 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc 1081 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga 1141 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa 1201 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga 1261 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca 1321 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat 1381 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa 1441 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt 1501 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg 1561 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt 1621 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg 1681 gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac 1741 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag 1801 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc 1861 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta 1921 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag 1981 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga 2041 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg 2101 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc 2161 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc 2221 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag 2281 acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca 2341 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc 2401 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta 2461 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt 2521 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc 2581 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg 2641 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg 2701 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag 2761 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc 2821 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat 2881 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg 2941 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc 3001 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt 3061 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca 3121 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt 3181 atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc 3241 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac 3301 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt 3361 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag 3421 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc 3481 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg 3541 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt 3601 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac 3661 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat 3721 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt 3781 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca 3841 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt 3901 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg 3961 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag 4021 gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc 4081 gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa 4141 taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat 4201 gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag 4261 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct 4321 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt 4381 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 4441 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca 4501 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc 4561 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga 4621 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc 4681 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca 4741 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca 4801 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt 4861 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg 4921 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat 4981 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa 5041 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga 5101 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc 5161 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa 5221 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca 5281 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg 5341 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca 5401 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg 5461 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa 5521 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc 5581 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt 5641 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg 5701 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg 5761 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg 5821 gcgtagagga tcgagatctc gatcccgcga aat //