LOCUS       sequence-205520-        4757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses thermostable variant of human carbonic anhydrase II with 
            TAG stop codon at site 233 and C-terminal 6x his tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4757)
  AUTHORS   Mehl R, van Zee K, Kean K
  TITLE     
  JOURNAL   2020
REFERENCE   2  (bases 1 to 4757)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "2020"
COMMENT     Sequence Label: pBad-CA TAG233
FEATURES             Location/Qualifiers
     source          1..4757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        15..299
                     /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)"
     primer_bind     222..241
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1113..1130
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1172..1189)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1342..1428
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1520..1547
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1566..1657
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1658..1726,1727..2518)
                     /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(1876..1895)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2689..3277
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3178..3197
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3431..3448
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3463..3603
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3529..3548)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3689..3711
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3867..4745)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 cccgccattc agagaagaaa ccaattgtcc atattgcatc agacattgcc gtcactgcgt
       61 cttttactgg ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa gcattctgta
      121 acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa agtgtctata atcacggcag
      181 aaaagtccac attgattatt tgcacggcgt cacactttgc tatgccatag catttttatc
      241 cataagatta gcggatccta cctgacgctt tttatcgcaa ctctctactg tttctccata
      301 cccgtttttt gggctaacag gaggaattaa ccatggcgca tcattggggt tacggtaaac
      361 acaacggtcc ggagcattgg cacaaagatt ttccaattgc gaagggcgaa cgtcaaagcc
      421 cggttgacat tgatacgcac acggcaaagt acgacccgag cctgaaaccg ctgagcgttt
      481 cctatgacca ggctacgagc ctgcgtatcc tgaacaatgg ccacaccttc aacgtggagt
      541 ttgatgattc ccaagataag gcggttctga aaggtggtcc gttggatggc acctaccgcc
      601 tgatccaatt tcactttcac tggggtagcc acgacggtca gggcagcgag cataccgtgg
      661 acaaaaagaa gtatgcagcc gaactgcacc tggtgcattg gaacacgaag tacggcgact
      721 tcggtaaagc ggtccagcaa ccggacggtc tggctgttct gggtattttc ctgaaggtcg
      781 gcagcgcgaa cccgggtctg cagaaagtgg ttgacgtgtt ggactctatc aagaccaaag
      841 gcaagagcgc ggacttcacc aatttcgatc cgcgtggtct gctgccggag agcctggatt
      901 actggactta tccgggcagc ctgaccaccc cgccattgct ggagtgcgtg acctggatcg
      961 tcttgaaaga accgatcagc gttagctctg aacaggtcag caagttccgc aagctgaatt
     1021 tcaatggtta gggcgagccg gaagaaccga tggtcgataa ttggcgtcct acccaaccgc
     1081 tgaaaaaccg ccagattaaa gcatccttta agcatcacca ccaccatcac taaaagcttg
     1141 gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa
     1201 gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca
     1261 tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga
     1321 gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     1381 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
     1441 gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact
     1501 gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa
     1561 actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     1621 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     1681 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     1741 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     1801 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     1861 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     1921 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     1981 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     2041 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     2101 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     2161 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     2221 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     2281 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     2341 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     2401 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     2461 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     2521 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     2581 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     2641 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2701 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2761 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2821 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2881 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2941 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3001 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3061 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3121 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3181 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3241 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3301 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3361 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3421 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     3481 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     3541 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     3601 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     3661 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     3721 tcatcaccga aacgcgcgag gcagcagatc aattcgcgcg cgaaggcgaa gcggcatgca
     3781 taatgtgcct gtcaaatgga cgaagcaggg attctgcaaa ccctatgcta ctccgtcaag
     3841 ccgtcaattg tctgattcgt taccaattat gacaacttga cggctacatc attcactttt
     3901 tcttcacaac cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc
     3961 gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg
     4021 gtggtgctca aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg
     4081 ctaatcccta actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc
     4141 tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa
     4201 gcctcgcgta cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc
     4261 cgcagtaaca attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct
     4321 tgcccggcgt taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc
     4381 gggcgaaaga accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag
     4441 gcgcgcggac gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg
     4501 tagtgatgaa tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct
     4561 cgtccctgat ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc
     4621 attcccagcg gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa
     4681 cccgccacca gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca
     4741 gccatacttt tcatact
//
