LOCUS       sequence-262980-        9560 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pET14b carrying the dabA2 gene (Uniprot: D0KWS7) with a C351A 
            mutation fused to a c-terminal strep tag and the dabB2 gene 
            (Uniprot: D0KWS8) with a c-terminal sfGFP V206K fusion and 6xHis 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9560)
  AUTHORS   Desmarais JJ, Flamholz AI, Blikstad C, Dugan EJ, Laughlin TG, 
            Oltrogge LM, Chen AW, Wetmore K, Diamond S, Wang JY, Savage DF
  TITLE     DABs are inorganic carbon pumps found throughout prokaryotic phyla.
  JOURNAL   Nat Microbiol. 2019 Aug 12. pii: 10.1038/s41564-019-0520-8. doi: 
            10.1038/s41564-019-0520-8.
  PUBMED    31406332
REFERENCE   2  (bases 1 to 9560)
  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: "Nat 
            Microbiol. 2019 Aug 12. pii: 10.1038/s41564-019-0520-8. doi: 
            10.1038/s41564-019-0520-8."
COMMENT     Sequence Label: pET14b-dabAB2 C351A
FEATURES             Location/Qualifiers
     source          1..9560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     910..929
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1013..1032
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1013..1031
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1063..1085
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3471..3488
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6027..6050
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(6102..6120)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6116..6163
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(6529..6534,6535..6551,6552..6557))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(6584..6602)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6670..6774
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6775..6843,6844..7635)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6993..7012)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7806..8394
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8295..8314
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8548..8565
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8580..8719
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8645..8664)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8805..8827
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8821..9012)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tggtttgcgc attcacagtt ctccgcaaga attgattggc tccaattctt ggagtggtga
       61 atccgttagc gaggtgccgc cggcttccat tcaggtcgag gtggcccggc tccatgcacc
      121 gcgacgcaac gcggggaggc agacaaggta tagggcggcg cctacaatcc atgccaaccc
      181 gttccatgtg ctcgccgagg cggcataaat cgccgtgacg atcagcggtc caatgatcga
      241 agttaggctg gtaagagccg cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac
      301 ctgcctggac agcatggcct gcaacgcggg catcccgatg ccgccggaag cgagaagaat
      361 cataatgggg aaggccatcc agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc
      421 gtcggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc
      481 agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat
      541 catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac
      601 ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc
      661 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgacgctc
      721 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
      781 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
      841 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
      901 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
      961 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga
     1021 ctcactatag ggacaccaca acggtttccc tctagaaata attttgttta actttaagaa
     1081 ggagatatac catgtccatg actatagagt tcagccacac aaccggtctt ctcctgctgg
     1141 ccatgcccgc actgctcctt atggccgctg taatcaaacc caagcaaggt gctgcttatg
     1201 cgcgcgctta tcggcaacgg gttcaatgga ccagttttgc ggcctttggc ctggccctgc
     1261 tcgctgtcgt cagttttcta tttgcaaggc aacagaatct gatgctcggt gccggtagcc
     1321 tccctgccgg attgggatta ttggcgcttt caattcaagt caatggcctg actctggtgc
     1381 tggcaagtct ggttagtttt gtgcttagcg tgattgcccg ctacagcgtg cagtatctcg
     1441 atggtgatcc ccagcaggcc cgctttttcc gcttgctggc ggtcaccggc ggatttttcc
     1501 tgttggtcgt catatcaggc aatttgggat tattcaccct cgccatcatt gcgaccggtt
     1561 ttggtttgca tcgactgctt tctttttatg ccgatcggcc gcgcgcgatc atggccacac
     1621 ataaaaaatc aattttcagc cggactgccg acgccctttt gctcgcggcc accgtcttga
     1681 tcggtcatca aatcggtagt ttggagttca gccaaatcag cgcctatgtt catgcgcaag
     1741 accacctatc cattgcgttg cacgtggctg cttggctgat cgtgctggcc gccatcctta
     1801 aaagcgcgca atttccgttt catggctggc tgattcaggt catggaagca cccacgccgg
     1861 tttccgcgct catgcacgcc ggtgtcgttt acagcggggc aatcattgtg ctgcgcacca
     1921 gcgaattatt ggccgccgac ggtaccgccc tgttgcttct ggccctgatt ggtttgatga
     1981 ccttggctat cggctcgttg gtcatgctga cccaatcggc gatcaaatca tcactggcct
     2041 ggtctaccgc cgctcagctc ggcttcatga tgctcgaact gggtttgggg ttgtttggct
     2101 tggccttgct gcacttggtt ggccactcgc tctacaaggc ccatgcgttc ttgtcctcgg
     2161 gcagcatgac cgatcacctg cgtcaggcca aagtgctcaa aaaccgccca atctctgttg
     2221 tggcctggtt taccacggtg attgtgagcg gcttgttcac cctcggtatc gcggcggcga
     2281 tgggtttatc gatcgatcag gagccgatgt tgcccgccgt gctgaccatc atcgccttgg
     2341 cgaccgctca gttgatgctc aaggcattga gccatggcac atggcgtgaa attctggtcg
     2401 ccgcaggcgc ggcaatcgcc atgaccggcg tgtatgtctt cctgcatgag gtgttcatca
     2461 caggctttgc cgacacgctg gccgcgacac cgcaacgtgc gccgctgctc gatctgctgt
     2521 tgatggcgat cacgatcata accttcctgt ttgttgcctg gcttcaaggc cccggcaaaa
     2581 ccttgatgag cccggagcga caattcgcct tgtttgttca cctcaataac gggctgtatc
     2641 tcgaccgctg ggtcgaacgc ttggccttcc gtttctggcc ggagaaagtt ggccgcgcac
     2701 cgaaaaaatc ctgcgccgtg attccgccta acccatccgg tatcgagccg tctggttctt
     2761 ctaaaggtga agaactgttc accggtgttg ttccgatcct ggttgaactg gatggtgatg
     2821 ttaacggcca caaattctct gttcgtggtg aaggtgaagg tgatgcaacc aacggtaaac
     2881 tgaccctgaa attcatctgc actaccggta aactgccggt tccatggccg actctggtga
     2941 ctaccctgac ctatggtgtt cagtgttttt ctcgttaccc ggatcacatg aagcagcatg
     3001 atttcttcaa atctgcaatg ccggaaggtt atgtacagga gcgcaccatt tctttcaaag
     3061 acgatggcac ctacaaaacc cgtgcagagg ttaaatttga aggtgatact ctggtgaacc
     3121 gtattgaact gaaaggcatt gatttcaaag aggacggcaa catcctgggc cacaaactgg
     3181 aatataactt caactcccat aacgtttaca tcaccgcaga caaacagaag aacggtatca
     3241 aagctaactt caaaattcgc cataacgttg aagacggtag cgtacagctg gcggaccact
     3301 accagcagaa cactccgatc ggtgatggtc cggttctgct gccggataac cactacctgt
     3361 ccacccagtc taaactgtcc aaagacccga acgaaaagcg cgaccacatg gtgctgctgg
     3421 agttcgttac tgcagcaggt atcacgcacg gcatggatga gctctacaaa caccatcatc
     3481 atcatcatta aatccctgcc acaaaggcac acaccattta cccgaggatt tcgaccatga
     3541 cgaccctgac atcattacaa cgcagcgaag cgcaacgaaa ccacattgtt gacctgattg
     3601 acaaagcctg cctgcgcatc gcgcccatct ggccactgga ttcgtttgtc gccgtaaacc
     3661 cctatcttgg tttgattgat caaccctttg atacggtggg tcggtatctg gaacaaaccg
     3721 tgggcgaatc cctatttatg gatcacggct ggtttgccga caagatcgcc caaggggaaa
     3781 tcaccgatga cgatttggcg caggcggcgc agcaactgga cccatcaatt agccttgata
     3841 ccatcaagca gcaactggcc gtccatcgac aacctgcacc cgcgctgcct ttggtcacaa
     3901 acgagctcga tcgccgcgac gcgccgccag tatctgaatt tgtcatcgag caagtcagcc
     3961 agttcatggc caactactat gatcgcgggc aagccctctg gcacttaccc aaagaagcca
     4021 gtgccagttt attcgcccaa tggcgccgct acaccttgat tgaccgcagc gccagcgccg
     4081 tcggcttaaa gcaggttcgc caacacttgc ttgccgtccc aagcgatgcc attgatgcat
     4141 tgttctgggc gctggaccag atcaatctgc ccgaaagtcg cctgcccgac tacctgttca
     4201 ccttattaaa aaccatcggt ggctgggcga gctggtgccg ctatctacac tttcaagccg
     4261 gtttgcacgg cgagtcgcaa cacgatttac gcgatttgct gatcattcgc ttggtctggg
     4321 ttgctttggt gatcaaagaa accagctcgg ctggacggca acaatggcgc gcgaagctaa
     4381 acgattggtt tgatccggcc aaattggtcg caagcccttc ggcgacagct actgcttcga
     4441 ctaaggctca atccagccgg atcgatgaaa tcctgctggc ggccgccgag caggcgtttc
     4501 gacgccggat caatgccggc ttgaaccgcc agcctgccga tgcacccgat cagcaagccg
     4561 aacgcccgac tgttcaggcc gcgtttgcca ttgacgtgcg ttccgaggtg ttccgccgcc
     4621 acctcgaagc cagcagcccg gggctcgaaa ccatcggttt tgccggtttt ttcggcctgc
     4681 cgattgatta ttgccgcatg ggcgaatcag aggctcggct acaaaacccg gtgctcatca
     4741 accccgccta ccgtgcgcag gagacgggcg atccggccat tgcccaacat cgccatgccc
     4801 gacaaagccg aggggccatc tggaagcaat tcaaactttc cgccgcctcg tgcttcacct
     4861 ttgtcgaatc cgcaggcttg agctacgtgc cgcgtttgct ggccgattca ctgggttggc
     4921 atcgctcatc gctgccgcct gatgcccccg ggcttacccc tgaggagcgt gcgcgtttgc
     4981 atccccaatt ggtcaaactc gatggcggcg cactatcgac gcaggaaaaa gtcgatctgg
     5041 ccgaaaaagt actccgcggg ctggggttaa cgcacacctt tgcgcccatc gtcctgctcg
     5101 ccggacacgg cagctccacc accaacaacc cgcatcgcgc ggggttggat tgcggtgcct
     5161 gcgccggtca ggcaggcgat gtgaatgctc gcgtcgcggt gcagttgctc aatgaggccg
     5221 ccgtgcgact ggggctgatc gaacggggta ttgccatccc gcgtgacacc cggttcgtgg
     5281 ccgccctgca cgacaccacc acagaccaca tcgaactact ggatttggat caatcaggca
     5341 tcgagagtga tcaattgtca tccttgaccc aggcgctcaa acaagccggt gagctcactc
     5401 gtctggagcg tttggtgacg cttgaggccc aagtcgatac cgttgatgcc gaaaaacagg
     5461 ccacgttccg agggcgagac tggtcgcaag tacggccgga atgggggcta gccggcaatg
     5521 ctgccttcat cgccgcaccc cgctggcgca cgcgcgggtt ggatcttggc ggccgcgcct
     5581 tcctgcatga ttacgattgg cgacacgata aggagtttgg cgttttgaac gtcatcatga
     5641 ccgcgccctt gatcgtcgca aactggatca acctgcaata ctacggctcc acggtcgata
     5701 atctgcatca aggggcgggc aataaagtgc tgcataatgt ggtcggcggc accgtcgggg
     5761 tgattgaagg caatggcggc gacctgcgag ttggtcttgc catgcaatcg ctgcacgatg
     5821 gtgaacagtg gcggcacgaa ccgctgcggc tctcggcgta cattgaggcg cccattgcag
     5881 aaatcgataa aattattgcc ggtcacgaca tgctgaacgc gttaatcaac aaccgctgga
     5941 tgcacatcct gcatatcgat gacaacggca tcccgcatcg ccgtcatgcc cacggtgact
     6001 ggcgaccgga accgatctct ggttcttgga gtcatcccca gttcgagaaa taaatccggc
     6061 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     6121 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     6181 atccggatat ccacaggacg ggtgtggtcg ccatgatcgc gtagtcgata gtggctccaa
     6241 gtagcgaagc gagcaggact gggcggcggc caaagcggtc ggacagtgct ccgagaacgg
     6301 gtgcgcatag aaattgcatc aacgcatata gcgctagcag cacgccatag tgactggcga
     6361 tgctgtcgga atggacgata tcccgcaaga ggcccggcag taccggcata accaagccta
     6421 tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg ttagatttca
     6481 tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat taaagcttat
     6541 cgatgataag ctgtcaaaca tgagaattct tgaagacgaa agggcctcgt gatacgccta
     6601 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     6661 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6721 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6781 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     6841 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     6901 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6961 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     7021 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     7081 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     7141 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     7201 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     7261 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     7321 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     7381 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     7441 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg ctctcgcggt
     7501 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     7561 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     7621 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     7681 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     7741 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7801 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     7861 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     7921 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     7981 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     8041 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     8101 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     8161 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     8221 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     8281 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     8341 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     8401 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     8461 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     8521 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     8581 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc
     8641 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     8701 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     8761 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gcctccggga gctgcatgtg
     8821 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     8881 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     8941 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     9001 ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc
     9061 gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact
     9121 ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac
     9181 tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca
     9241 gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag
     9301 actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt
     9361 gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag
     9421 gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg
     9481 ccaggaccca acgctgcccg agatgcgccg cgtgcggctg ctggagatgg cggacgcgat
     9541 ggatatgttc tgccaagggt
//
