LOCUS       sequence-205515-        4757 bp    DNA     circular SYN 15-DEC-2025
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    .
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-CAts TAG97
FEATURES             Location/Qualifiers
     source          1..4757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..300
                     /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     223..242
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1114..1131
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1173..1190)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1343..1429
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1521..1548
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1567..1658
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1659..1727,1728..2519)
                     /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(1877..1896)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2690..3278
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3179..3198
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3432..3449
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3464..3604
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3530..3549)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3690..3712
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3868..4746)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 tcccgccatt cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg
       61 tcttttactg gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt
      121 aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca
      181 gaaaagtcca cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat
      241 ccataagatt agcggatcct acctgacgct ttttatcgca actctctact gtttctccat
      301 acccgttttt tgggctaaca ggaggaatta accatggcgc atcattgggg ttacggtaaa
      361 cacaacggtc cggagcattg gcacaaagat tttccaattg cgaagggcga acgtcaaagc
      421 ccggttgaca ttgatacgca cacggcaaag tacgacccga gcctgaaacc gctgagcgtt
      481 tcctatgacc aggctacgag cctgcgtatc ctgaacaatg gccacacctt caacgtggag
      541 tttgatgatt cccaagataa ggcggttctg aaaggtggtc cgttggatgg cacctaccgc
      601 ctgatccaat ttcactttca ctagggtagc cacgacggtc agggcagcga gcataccgtg
      661 gacaaaaaga agtatgcagc cgaactgcac ctggtgcatt ggaacacgaa gtacggcgac
      721 ttcggtaaag cggtccagca accggacggt ctggctgttc tgggtatttt cctgaaggtc
      781 ggcagcgcga acccgggtct gcagaaagtg gttgacgtgt tggactctat caagaccaaa
      841 ggcaagagcg cggacttcac caatttcgat ccgcgtggtc tgctgccgga gagcctggat
      901 tactggactt atccgggcag cctgaccacc ccgccattgc tggagtgcgt gacctggatc
      961 gtcttgaaag aaccgatcag cgttagctct gaacaggtca gcaagttccg caagctgaat
     1021 ttcaatggtg agggcgagcc ggaagaaccg atggtcgata attggcgtcc tacccaaccg
     1081 ctgaaaaacc gccagattaa agcatccttt aagcatcacc accaccatca ctaaaagctt
     1141 ggctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga
     1201 agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc
     1261 atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg
     1321 agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     1381 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc
     1441 ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac
     1501 tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca
     1561 aactcttttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1621 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     1681 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     1741 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     1801 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     1861 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     1921 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     1981 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     2041 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     2101 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     2161 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     2221 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     2281 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     2341 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     2401 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     2461 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     2521 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     2581 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     2641 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2701 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2761 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2821 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2881 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2941 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3001 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3061 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3121 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3181 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3241 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3301 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3361 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3421 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     3481 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct
     3541 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     3601 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3661 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3721 gtcatcaccg aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc
     3781 ataatgtgcc tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa
     3841 gccgtcaatt gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt
     3901 ttcttcacaa ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg
     3961 cgagaaatag agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg
     4021 ggtggtgctc aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac
     4081 gctaatccct aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg
     4141 ctgtgcgacg ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca
     4201 agcctcgcgt acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg
     4261 ccgcagtaac aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc
     4321 ttgcccggcg ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc
     4381 cgggcgaaag aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta
     4441 ggcgcgcgga cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc
     4501 gtagtgatga atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc
     4561 tcgtccctga tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt
     4621 cattcccagc ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa
     4681 acccgccacc agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc
     4741 agccatactt ttcatac
//
