LOCUS Exported 8635 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses machinery necessary for selenocysteine incorporation at UAG codons. ACCESSION . VERSION . KEYWORDS pSecUAG-Evol2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8635) AUTHORS Mukai T, Sevostyanova A, Suzuki T, Fu X, Soll D TITLE A Facile Method for Producing Selenocysteine-Containing Proteins. JOURNAL Angew Chem Int Ed Engl. 2018 Jun 11;57(24):7215-7219. doi: 10.1002/anie.201713215. Epub 2018 May 9. PUBMED 29631320 REFERENCE 2 (bases 1 to 8635) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8635 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 59..78 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" primer_bind complement(316..333) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 486..572 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" promoter complement(3998..4045) /label=EM7 promoter /note="synthetic bacterial promoter " CDS complement(5300..6115) /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 6023..6042 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" promoter complement(6116..6207) /gene="bla" /label=AmpR promoter primer_bind complement(6473..6492) /label=pBR322ori-F /note="pBR322 origin, forward primer" misc_feature 7178..7318 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(7244..7263) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7404..7426 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(7582..8460) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" promoter 8487..136 /gene="araBAD" /label=araBAD promoter /note="promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction (Guzman et al., 1995)" ORIGIN 1 gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat 61 gccatagcat ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc 121 tctactgttt ctccataccc gtttgaattc ggacgggggt tccgtctggt ggcggtcgcg 181 ggctctaaac ccgtcaggac gctgtgcagg cgttaggttc gattcctccc ccgtccgcca 241 ggatctagag tcgacctgca gatccttagc gaaagctaag gatttttttt aggaattaac 301 catggatccg agcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca 361 gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt 421 gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca 481 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt 541 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga 601 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta 661 agcagaaggc catcctgacg gatggccgcg gccgcctcta aacgggtctt gaggggtttt 721 ttgctgaaac ctcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa 781 ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga accgacgaca 841 agctgacgac cgggtctccg caagtggcac ttttcgggga aatgtgcgcg gaacccctat 901 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaga tatgctttga 961 aacaaagttt acaaggctct tcttatccat atagcctgtg tgcctgtcca cctcttttcc 1021 gtcttttaaa acgatgaggg taggtattga cataaactta aattttactg ccaattcacg 1081 tgcattatcc acgttagact gtgctatcac agccttgtcg ccgagttccg cctcggcagc 1141 ctgcagctca ggactgagct gtacgcatcc cggctaccac ggtgcccaaa agtcaattaa 1201 aacgggcttg gcggtcttaa cggtttcatc aaaattagca tttgtaatat ccaatactgc 1261 cataatcatg gtctgtttcc tgtgtgatca ttggataacc tcgataaact ggtttccggt 1321 ataggctttc agctggccta tgggggagag ggtcagcccc gcttgtgtcg cgatagcaag 1381 gagctcggct tcactttctt taccgacggc aaccagcaag ccgccgctgg tctgcggatc 1441 gcacatgatg ttgcgggtgc gttcatccat ggcaccgagc ttggcgccat aggaatcgaa 1501 gttgcgcagg gtaccgcccg gtacgcagcc ctcggacagg tagtaatcta cttcgtcgag 1561 cagtggcagc gccttgaaat cgagggtggc acacacccct gagccttcgc acatctcaag 1621 caggtgtccc gccaggccaa acccggtcac atccgtcatg gcgtgcacgc cgggcagttc 1681 ggcaaagcgc tggccaatct tgttgagggt gcacatggcg ttgggggcca gctgctcatg 1741 ctctggcttc aatttgccct tcttctgggc cgtggtgagg atgccgatac cgaggggctt 1801 ggtcaggtag aggatgtcac ccgcctgggc cgtgtcgttc tgcttgatgg cattgagcgg 1861 cactataccg gtcacagcaa gaccgaagat gggctcgggg gcatcgatac tgtggccgcc 1921 agccaaggat atgcccgctt catggcacac ctggcggccg ccatctatca cctgctgggc 1981 cacttccggg gctagggtgt tgatgggcca gccaaggatg gcaatggcaa cgatgggctt 2041 gccgcccatg gcgtagatgt cgctgatggc gttggtggcc gcgatgcggc caaaggtaaa 2101 gggatcatcg acgatgggca tgaagaagtc ggtggtggaa acaatgccct gaccgttgcc 2161 gatatcgacc acggccgcgt catccttgct gctgttgcca accaccaggg tcgggtcgtc 2221 aaagcccggg atctggctct tgagaatggt gtcgagcacc ttgggagaaa ttttgcagcc 2281 gcagccagcc ccgtggctgt attgggtcag acgaatggaa gacacgatta ccccttgttt 2341 ggctgtttct caagatgaaa cagcgtatat caggcaaaag gagatgaccc tgagcgggcc 2401 attggacagg gcattatgcc acaaggactc tgcgggttcg aatcacaata gcctgtcgaa 2461 aatgccgcca gcggaactgg cggctgtggg atcagggctc ctcggtcgca gtggtcggag 2521 tgggcagtag caactccttg agttgggcga taagcagtgc aatctcggtc ggcagcaggg 2581 tcgccatatt gagcagcacc ttctgctggc gcacggtggc gatgaccggc accggcagtt 2641 tccgcagggc atcgagcagc tgctgggccg gacgcgggtc ggtgcattcg agcgcaggcg 2701 cagggtagaa ctcgtccggc agggtgccac cacccaccac cagctgggcg gggaccggca 2761 caaagcagcc gggcagggcg gccatcagct gatcggcgcg ggcctgcatg gcggcagggt 2821 tgctcaaggt gcgctgggcg atgccctcgc cgatggggga cttgttgagc ttgtggatga 2881 gcaggcgttc cagcagggag tagacgatgc ggctcgggcg gaaggtgcgc atcatggggt 2941 gtttttccag ccgcttgatg aggtcgctgc ggccgctgat gatgcccgat tgcgggccac 3001 ccagcagctt gtcgccggag tagcagacca gatccgcccc cgccttgatg tactgacgca 3061 ccgaggtttc gtccggtgca aactcctcgg tggtcaagcc cgagccctga tccaccgcca 3121 gcaccacgtg ctcgggcagg gcgcgggcca cctcgccaat atcgggggat tcggtaaagc 3181 cgcgaatggc gaaattggat ctgtgtacca tcagcaccag cgcggtctga tctgtgatgg 3241 catcgaggta atctttggcg gtagtgatat tggtggtgcc cacctccacc agtttggcgc 3301 cggagagcgc cagaatgtcg ggaatgcgaa agccgccacc aatctggatc tgttcgcccc 3361 gcgagacgat cacctcgcgc cccttggcta tctcctgcag cagcaagaag agcgaagcgg 3421 cgttgttgtt gaccaccagc gaatcctcgg cctgggtgag cacacggagc aggggggcga 3481 tcagcccctt gcgcccgccg cgcttgccgg tggcgagatc cagttccaga ttgttgtagc 3541 cagtgttgag gtcgcgcacc tcgtcccaca gctcgcgact tagcggcgag cgccccagat 3601 tggtgtgcac cagggtgccg gtggcgttga tcacccgggt ctgacgttgg cgcagctgct 3661 gctggcaacg cttggcaatc agtgcctcga tttgctcggg ggcaacccca tgctggcgaa 3721 atgcctcgct ctggcgcaat tcgctcagga catcgcgcac cgccgaggtc acaaaagggc 3781 ggctcagcgc ctcgataaaa ccggtgagaa agggttgctg cagcagctgt tccacttgcg 3841 gtagacggcg cgcttgttgc tggctgggct gtggcagtga atctggcagt gaatctggca 3901 gtgaatcgtc ggcagtggga catgattcgg gctgactgtg agagtgggcg atggctggcg 3961 cgtgagacga gttcgtcacg gtttagttcc tcaccttgtc gtattatact atgccgatat 4021 actatgccga tgattaattg tcaacacgtg attatcccag caaacggtga atacgttgca 4081 ggccggtcac cagacgatcc acttcttcat gggtgttata catggccagc gacgcccgac 4141 acatcgcagg gacgttgtaa taggccatca atggcattgc ggcgtgatgt ccggtacgca 4201 cagcaatgcc gtaattatcg agaaaactgc caacatcata ggcgtggtgt ttaccgagat 4261 taaaagcaat aacgccaagc ctgttttgtg ggccatagag agtgagatcc ggtacagatt 4321 ccagctgtga tagcgcataa tgcatcagat tctgttcata ctcggctatg ttattaagcc 4381 ccagcgccga aacatactcc agcgccgcgc caagaccaat gatgcccccg gtattgggtg 4441 taccggcttc aaaccgccat ggtgctttgg tccaggtagt gccttcactc aggctgacgg 4501 tggcgatcat agaaccgccc ccttcccacg gcggcatctc ctgcaacaag gcttctttca 4561 cataaagaat gccaattccg gtggggccat acagtttatg cccggagaac acgtaaaagt 4621 cgcaatccag cgcctgaaca tccaccggat gatgcatcac cgcctgagcg ccatccacca 4681 gcacttttgc gccatgctgg tgcgcaagcg tgatcatttc cgccagtgga ttttctgtgc 4741 caagcacgtt ggagacatga gtaattgcca gcaggcgagt tttctcatca aacagcgtag 4801 gcagcgtctc cagttgcaac gtaccatcgg gattgagcgg gatcacacgc agctctgcgc 4861 caacgcgtgc gcaaagcatc tgccagggaa caatgttagc gtggtgctcc atctgactga 4921 tgatgatgtt atcgcccgcc cgcacgttgc tgttgcccca gctattggcg accagattga 4981 tcccttccgt cgtgccgcgg acgaacacca gctcttccgc cgaacgggca ttaataaaca 5041 gcgatgcccg cttgcgcacg ttctccattt tctcggtcgc ctgggcgctt aaggtatgaa 5101 taccacgatg caccgccgcg tagccatgac gataaaactc ggcctcggcg tcaatcacct 5161 ggctcggttt ctgcgcactg gcggcgctgt cgagataagc cagcggcaaa ccgtttacct 5221 cacgcgaaag caccggaaag tcggcccgca ctttgtcgac ggaaaaaatc atcttgcacc 5281 tcctggcagt tattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat 5341 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa 5401 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc 5461 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga 5521 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc 5581 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac 5641 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcggtcgctg ttaaaaggac 5701 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat 5761 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag 5821 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca 5881 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac 5941 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg 6001 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca 6061 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcatactct 6121 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 6181 ttgaatgtat ttagaaaaat aaacaaatag gcatgcagcg ctcttccgct tcctcgctca 6241 ctgactcgct acgctcggtc gttcgactgc ggcgagcggt gtcagctcac tcaaaagcgg 6301 taatacggtt atccacagaa tcaggggata aagccggaaa gaacatgtga gcaaaaagca 6361 aagcaccgga agaagccaac gccgcaggcg tttttccata ggctccgccc ccctgacgag 6421 catcacaaaa atcgacgctc aagccagagg tggcgaaacc cgacaggact ataaagatac 6481 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 6541 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 6601 tggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 6661 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 6721 cacgacttat cgccactggc agcagccatt ggtaactgat ttagaggact ttgtcttgaa 6781 gttatgcacc tgttaaggct aaactgaaag aacagatttt ggtgagtgcg gtcctccaac 6841 ccacttacct tggttcaaag agttggtagc tcagcgaacc ttgagaaaac caccgttggt 6901 agcggtggtt tttctttatt tatgagatga tgaatcaatc ggtctatcaa gtcaacgaac 6961 agctattccg ggccggccgg agcctatgga aaaacgccag caacgcggcc tttttacggt 7021 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg 7081 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg 7141 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta 7201 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg 7261 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc 7321 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc 7381 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc 7441 accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc atgcataatg 7501 tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc 7561 aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc 7621 acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa 7681 atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt 7741 gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat 7801 ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc 7861 gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc 7921 gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag 7981 taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc 8041 ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg 8101 aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg 8161 cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg 8221 atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc 8281 ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc 8341 cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc 8401 caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat 8461 acttttcata ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg 8521 ccgtcactgc gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa 8581 aagcattctg taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagt //