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 97 and C-terminal 6x his tag.
ACCESSION   .
VERSION     .
KEYWORDS    pBad-CAts TAG97
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  JOURNAL   Unpublished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 tccaatttca ctttcactag 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
//
