LOCUS Exported 4757 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION expresses thermostable variant of human carbonic anhydrase II with TAG stop codon at site 93 and C-terminal 6x his tag. ACCESSION . VERSION . KEYWORDS pBad-CA TAG93 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4757) TITLE Bacterial expression plasmids for genetic code expansion laboratory course JOURNAL Unpublished REFERENCE 2 (bases 1 to 4757) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4757 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 189..208 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" CDS 1080..1097 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1139..1156) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 1309..1395 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 1487..1514 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 1533..1624 /gene="bla" /label=AmpR promoter CDS 1625..2485 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1843..1862) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2656..3244 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3145..3164 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3398..3415 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3430..3570 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3496..3515) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3656..3678 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3834..4712) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" promoter 4739..266 /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 ttgcatcaga cattgccgtc actgcgtctt ttactggctc ttctcgctaa ccaaaccggt 61 aaccccgctt attaaaagca ttctgtaaca aagcgggacc aaagccatga caaaaacgcg 121 taacaaaagt gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac 181 actttgctat gccatagcat ttttatccat aagattagcg gatcctacct gacgcttttt 241 atcgcaactc tctactgttt ctccataccc gttttttggg ctaacaggag gaattaacca 301 tggcgcatca ttggggttac ggtaaacaca acggtccgga gcattggcac aaagattttc 361 caattgcgaa gggcgaacgt caaagcccgg ttgacattga tacgcacacg gcaaagtacg 421 acccgagcct gaaaccgctg agcgtttcct atgaccaggc tacgagcctg cgtatcctga 481 acaatggcca caccttcaac gtggagtttg atgattccca agataaggcg gttctgaaag 541 gtggtccgtt ggatggcacc taccgcctga tccaatagca ctttcactgg ggtagccacg 601 acggtcaggg cagcgagcat accgtggaca aaaagaagta tgcagccgaa ctgcacctgg 661 tgcattggaa cacgaagtac ggcgacttcg gtaaagcggt ccagcaaccg gacggtctgg 721 ctgttctggg tattttcctg aaggtcggca gcgcgaaccc gggtctgcag aaagtggttg 781 acgtgttgga ctctatcaag accaaaggca agagcgcgga cttcaccaat ttcgatccgc 841 gtggtctgct gccggagagc ctggattact ggacttatcc gggcagcctg accaccccgc 901 cattgctgga gtgcgtgacc tggatcgtct tgaaagaacc gatcagcgtt agctctgaac 961 aggtcagcaa gttccgcaag ctgaatttca atggtgaggg cgagccggaa gaaccgatgg 1021 tcgataattg gcgtcctacc caaccgctga aaaaccgcca gattaaagca tcctttaagc 1081 atcaccacca ccatcactaa aagcttggct gttttggcgg atgagagaag attttcagcc 1141 tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca 1201 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg 1261 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga 1321 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc 1381 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg 1441 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg 1501 acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa 1561 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa 1621 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct 1681 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg 1741 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg 1801 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt 1861 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga 1921 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga 1981 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac 2041 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg 2101 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac 2161 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct 2221 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct 2281 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg 2341 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat 2401 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg 2461 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat 2521 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct 2581 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa 2641 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa 2701 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc 2761 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta 2821 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct 2881 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg 2941 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag 3001 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc 3061 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg 3121 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt 3181 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg 3241 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca 3301 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg 3361 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc 3421 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat 3481 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg 3541 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg 3601 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg 3661 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gcagatcaat 3721 tcgcgcgcga aggcgaagcg gcatgcataa tgtgcctgtc aaatggacga agcagggatt 3781 ctgcaaaccc tatgctactc cgtcaagccg tcaattgtct gattcgttac caattatgac 3841 aacttgacgg ctacatcatt cactttttct tcacaaccgg cacggaactc gctcgggctg 3901 gccccggtgc attttttaaa tacccgcgag aaatagagtt gatcgtcaaa accaacattg 3961 cgaccgacgg tggcgatagg catccgggtg gtgctcaaaa gcagcttcgc ctggctgata 4021 cgttggtcct cgcgccagct taagacgcta atccctaact gctggcggaa aagatgtgac 4081 agacgcgacg gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa attgctgtct 4141 gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc gattatccat cggtggatgg 4201 agcgactcgt taatcgcttc catgcgccgc agtaacaatt gctcaagcag atttatcgcc 4261 agcagctccg aatagcgccc ttccccttgc ccggcgttaa tgatttgccc aaacaggtcg 4321 ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaacc ccgtattggc aaatattgac 4381 ggccagttaa gccattcatg ccagtaggcg cgcggacgaa agtaaaccca ctggtgatac 4441 cattcgcgag cctccggatg acgaccgtag tgatgaatct ctcctggcgg gaacagcaaa 4501 atatcacccg gtcggcaaac aaattctcgt ccctgatttt tcaccacccc ctgaccgcga 4561 atggtgagat tgagaatata acctttcatt cccagcggtc ggtcgataaa aaaatcgaga 4621 taaccgttgg cctcaatcgg cgttaaaccc gccaccagat gggcattaaa cgagtatccc 4681 ggcagcaggg gatcattttg cgcttcagcc atacttttca tactcccgcc attcagagaa 4741 gaaaccaatt gtccata //