LOCUS       sequence-205516-        4757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses thermostable variant of human carbonic anhydrase II with 
            TAG stop codon at site 126 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 TAG126
FEATURES             Location/Qualifiers
     source          1..4757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..292
                     /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     215..234
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1106..1123
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1165..1182)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1335..1421
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1513..1540
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1559..1650
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1651..1719,1720..2511)
                     /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(1869..1888)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2682..3270
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3171..3190
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3424..3441
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3456..3596
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3522..3541)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3682..3704
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3860..4738)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 ttcagagaag aaaccaattg tccatattgc atcagacatt gccgtcactg cgtcttttac
       61 tggctcttct cgctaaccaa accggtaacc ccgcttatta aaagcattct gtaacaaagc
      121 gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc
      181 cacattgatt atttgcacgg cgtcacactt tgctatgcca tagcattttt atccataaga
      241 ttagcggatc ctacctgacg ctttttatcg caactctcta ctgtttctcc atacccgttt
      301 tttgggctaa caggaggaat taaccatggc gcatcattgg ggttacggta aacacaacgg
      361 tccggagcat tggcacaaag attttccaat tgcgaagggc gaacgtcaaa gcccggttga
      421 cattgatacg cacacggcaa agtacgaccc gagcctgaaa ccgctgagcg tttcctatga
      481 ccaggctacg agcctgcgta tcctgaacaa tggccacacc ttcaacgtgg agtttgatga
      541 ttcccaagat aaggcggttc tgaaaggtgg tccgttggat ggcacctacc gcctgatcca
      601 atttcacttt cactggggta gccacgacgg tcagggcagc gagcataccg tggacaaaaa
      661 gaagtatgca gccgaactgc acctggtgca ttggaacacg tagtacggcg acttcggtaa
      721 agcggtccag caaccggacg gtctggctgt tctgggtatt ttcctgaagg tcggcagcgc
      781 gaacccgggt ctgcagaaag tggttgacgt gttggactct atcaagacca aaggcaagag
      841 cgcggacttc accaatttcg atccgcgtgg tctgctgccg gagagcctgg attactggac
      901 ttatccgggc agcctgacca ccccgccatt gctggagtgc gtgacctgga tcgtcttgaa
      961 agaaccgatc agcgttagct ctgaacaggt cagcaagttc cgcaagctga atttcaatgg
     1021 tgagggcgag ccggaagaac cgatggtcga taattggcgt cctacccaac cgctgaaaaa
     1081 ccgccagatt aaagcatcct ttaagcatca ccaccaccat cactaaaagc ttggctgttt
     1141 tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct
     1201 gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa
     1261 ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg
     1321 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     1381 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     1441 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
     1501 atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt
     1561 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1621 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     1681 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1741 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1801 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1861 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     1921 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1981 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     2041 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     2101 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     2161 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     2221 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2281 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2341 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     2401 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     2461 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     2521 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2581 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2641 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2701 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2761 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2821 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2881 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2941 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3001 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3061 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3121 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3181 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3241 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3301 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3361 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3421 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     3481 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     3541 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     3601 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     3661 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     3721 cgaaacgcgc gaggcagcag atcaattcgc gcgcgaaggc gaagcggcat gcataatgtg
     3781 cctgtcaaat ggacgaagca gggattctgc aaaccctatg ctactccgtc aagccgtcaa
     3841 ttgtctgatt cgttaccaat tatgacaact tgacggctac atcattcact ttttcttcac
     3901 aaccggcacg gaactcgctc gggctggccc cggtgcattt tttaaatacc cgcgagaaat
     3961 agagttgatc gtcaaaacca acattgcgac cgacggtggc gataggcatc cgggtggtgc
     4021 tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc
     4081 ctaactgctg gcggaaaaga tgtgacagac gcgacggcga caagcaaaca tgctgtgcga
     4141 cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct gatgtactga caagcctcgc
     4201 gtacccgatt atccatcggt ggatggagcg actcgttaat cgcttccatg cgccgcagta
     4261 acaattgctc aagcagattt atcgccagca gctccgaata gcgcccttcc ccttgcccgg
     4321 cgttaatgat ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa
     4381 agaaccccgt attggcaaat attgacggcc agttaagcca ttcatgccag taggcgcgcg
     4441 gacgaaagta aacccactgg tgataccatt cgcgagcctc cggatgacga ccgtagtgat
     4501 gaatctctcc tggcgggaac agcaaaatat cacccggtcg gcaaacaaat tctcgtccct
     4561 gatttttcac caccccctga ccgcgaatgg tgagattgag aatataacct ttcattccca
     4621 gcggtcggtc gataaaaaaa tcgagataac cgttggcctc aatcggcgtt aaacccgcca
     4681 ccagatgggc attaaacgag tatcccggca gcaggggatc attttgcgct tcagccatac
     4741 ttttcatact cccgcca
//
