LOCUS       sequence-262984-        9560 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pET14b carrying the dabA2 gene (Uniprot: D0KWS7) with a D353A 
            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 D353A
FEATURES             Location/Qualifiers
     source          1..9560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1155..1174
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1258..1277
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1258..1276
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1308..1330
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3716..3733
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6272..6295
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(6347..6365)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6361..6408
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(6774..6779,6780..6796,6797..6802))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(6829..6847)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6915..7019
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7020..7088,7089..7880)
                     /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(7238..7257)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8051..8639
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8540..8559
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8793..8810
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8825..8964
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8890..8909)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9050..9072
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(9066..9257)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
       61 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
      121 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct
      181 gcccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca
      241 agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt
      301 ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc ccggctccat
      361 gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac aatccatgcc
      421 aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag cggtccaatg
      481 atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg atggtcgtca
      541 tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga
      601 agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa gacgtagccc
      661 agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg
      721 ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg
      781 ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc
      841 ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc
      901 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
      961 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
     1021 agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     1081 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     1141 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     1201 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa
     1261 tacgactcac tatagggaca ccacaacggt ttccctctag aaataatttt gtttaacttt
     1321 aagaaggaga tataccatgt ccatgactat agagttcagc cacacaaccg gtcttctcct
     1381 gctggccatg cccgcactgc tccttatggc cgctgtaatc aaacccaagc aaggtgctgc
     1441 ttatgcgcgc gcttatcggc aacgggttca atggaccagt tttgcggcct ttggcctggc
     1501 cctgctcgct gtcgtcagtt ttctatttgc aaggcaacag aatctgatgc tcggtgccgg
     1561 tagcctccct gccggattgg gattattggc gctttcaatt caagtcaatg gcctgactct
     1621 ggtgctggca agtctggtta gttttgtgct tagcgtgatt gcccgctaca gcgtgcagta
     1681 tctcgatggt gatccccagc aggcccgctt tttccgcttg ctggcggtca ccggcggatt
     1741 tttcctgttg gtcgtcatat caggcaattt gggattattc accctcgcca tcattgcgac
     1801 cggttttggt ttgcatcgac tgctttcttt ttatgccgat cggccgcgcg cgatcatggc
     1861 cacacataaa aaatcaattt tcagccggac tgccgacgcc cttttgctcg cggccaccgt
     1921 cttgatcggt catcaaatcg gtagtttgga gttcagccaa atcagcgcct atgttcatgc
     1981 gcaagaccac ctatccattg cgttgcacgt ggctgcttgg ctgatcgtgc tggccgccat
     2041 ccttaaaagc gcgcaatttc cgtttcatgg ctggctgatt caggtcatgg aagcacccac
     2101 gccggtttcc gcgctcatgc acgccggtgt cgtttacagc ggggcaatca ttgtgctgcg
     2161 caccagcgaa ttattggccg ccgacggtac cgccctgttg cttctggccc tgattggttt
     2221 gatgaccttg gctatcggct cgttggtcat gctgacccaa tcggcgatca aatcatcact
     2281 ggcctggtct accgccgctc agctcggctt catgatgctc gaactgggtt tggggttgtt
     2341 tggcttggcc ttgctgcact tggttggcca ctcgctctac aaggcccatg cgttcttgtc
     2401 ctcgggcagc atgaccgatc acctgcgtca ggccaaagtg ctcaaaaacc gcccaatctc
     2461 tgttgtggcc tggtttacca cggtgattgt gagcggcttg ttcaccctcg gtatcgcggc
     2521 ggcgatgggt ttatcgatcg atcaggagcc gatgttgccc gccgtgctga ccatcatcgc
     2581 cttggcgacc gctcagttga tgctcaaggc attgagccat ggcacatggc gtgaaattct
     2641 ggtcgccgca ggcgcggcaa tcgccatgac cggcgtgtat gtcttcctgc atgaggtgtt
     2701 catcacaggc tttgccgaca cgctggccgc gacaccgcaa cgtgcgccgc tgctcgatct
     2761 gctgttgatg gcgatcacga tcataacctt cctgtttgtt gcctggcttc aaggccccgg
     2821 caaaaccttg atgagcccgg agcgacaatt cgccttgttt gttcacctca ataacgggct
     2881 gtatctcgac cgctgggtcg aacgcttggc cttccgtttc tggccggaga aagttggccg
     2941 cgcaccgaaa aaatcctgcg ccgtgattcc gcctaaccca tccggtatcg agccgtctgg
     3001 ttcttctaaa ggtgaagaac tgttcaccgg tgttgttccg atcctggttg aactggatgg
     3061 tgatgttaac ggccacaaat tctctgttcg tggtgaaggt gaaggtgatg caaccaacgg
     3121 taaactgacc ctgaaattca tctgcactac cggtaaactg ccggttccat ggccgactct
     3181 ggtgactacc ctgacctatg gtgttcagtg tttttctcgt tacccggatc acatgaagca
     3241 gcatgatttc ttcaaatctg caatgccgga aggttatgta caggagcgca ccatttcttt
     3301 caaagacgat ggcacctaca aaacccgtgc agaggttaaa tttgaaggtg atactctggt
     3361 gaaccgtatt gaactgaaag gcattgattt caaagaggac ggcaacatcc tgggccacaa
     3421 actggaatat aacttcaact cccataacgt ttacatcacc gcagacaaac agaagaacgg
     3481 tatcaaagct aacttcaaaa ttcgccataa cgttgaagac ggtagcgtac agctggcgga
     3541 ccactaccag cagaacactc cgatcggtga tggtccggtt ctgctgccgg ataaccacta
     3601 cctgtccacc cagtctaaac tgtccaaaga cccgaacgaa aagcgcgacc acatggtgct
     3661 gctggagttc gttactgcag caggtatcac gcacggcatg gatgagctct acaaacacca
     3721 tcatcatcat cattaaatcc ctgccacaaa ggcacacacc atttacccga ggatttcgac
     3781 catgacgacc ctgacatcat tacaacgcag cgaagcgcaa cgaaaccaca ttgttgacct
     3841 gattgacaaa gcctgcctgc gcatcgcgcc catctggcca ctggattcgt ttgtcgccgt
     3901 aaacccctat cttggtttga ttgatcaacc ctttgatacg gtgggtcggt atctggaaca
     3961 aaccgtgggc gaatccctat ttatggatca cggctggttt gccgacaaga tcgcccaagg
     4021 ggaaatcacc gatgacgatt tggcgcaggc ggcgcagcaa ctggacccat caattagcct
     4081 tgataccatc aagcagcaac tggccgtcca tcgacaacct gcacccgcgc tgcctttggt
     4141 cacaaacgag ctcgatcgcc gcgacgcgcc gccagtatct gaatttgtca tcgagcaagt
     4201 cagccagttc atggccaact actatgatcg cgggcaagcc ctctggcact tacccaaaga
     4261 agccagtgcc agtttattcg cccaatggcg ccgctacacc ttgattgacc gcagcgccag
     4321 cgccgtcggc ttaaagcagg ttcgccaaca cttgcttgcc gtcccaagcg atgccattga
     4381 tgcattgttc tgggcgctgg accagatcaa tctgcccgaa agtcgcctgc ccgactacct
     4441 gttcacctta ttaaaaacca tcggtggctg ggcgagctgg tgccgctatc tacactttca
     4501 agccggtttg cacggcgagt cgcaacacga tttacgcgat ttgctgatca ttcgcttggt
     4561 ctgggttgct ttggtgatca aagaaaccag ctcggctgga cggcaacaat ggcgcgcgaa
     4621 gctaaacgat tggtttgatc cggccaaatt ggtcgcaagc ccttcggcga cagctactgc
     4681 ttcgactaag gctcaatcca gccggatcga tgaaatcctg ctggcggccg ccgagcaggc
     4741 gtttcgacgc cggatcaatg ccggcttgaa ccgccagcct gccgatgcac ccgatcagca
     4801 agccgaacgc ccgactgttc aggccgcgtt ttgcattgcg gtgcgttccg aggtgttccg
     4861 ccgccacctc gaagccagca gcccggggct cgaaaccatc ggttttgccg gttttttcgg
     4921 cctgccgatt gattattgcc gcatgggcga atcagaggct cggctacaaa acccggtgct
     4981 catcaacccc gcctaccgtg cgcaggagac gggcgatccg gccattgccc aacatcgcca
     5041 tgcccgacaa agccgagggg ccatctggaa gcaattcaaa ctttccgccg cctcgtgctt
     5101 cacctttgtc gaatccgcag gcttgagcta cgtgccgcgt ttgctggccg attcactggg
     5161 ttggcatcgc tcatcgctgc cgcctgatgc ccccgggctt acccctgagg agcgtgcgcg
     5221 tttgcatccc caattggtca aactcgatgg cggcgcacta tcgacgcagg aaaaagtcga
     5281 tctggccgaa aaagtactcc gcgggctggg gttaacgcac acctttgcgc ccatcgtcct
     5341 gctcgccgga cacggcagct ccaccaccaa caacccgcat cgcgcggggt tggattgcgg
     5401 tgcctgcgcc ggtcaggcag gcgatgtgaa tgctcgcgtc gcggtgcagt tgctcaatga
     5461 ggccgccgtg cgactggggc tgatcgaacg gggtattgcc atcccgcgtg acacccggtt
     5521 cgtggccgcc ctgcacgaca ccaccacaga ccacatcgaa ctactggatt tggatcaatc
     5581 aggcatcgag agtgatcaat tgtcatcctt gacccaggcg ctcaaacaag ccggtgagct
     5641 cactcgtctg gagcgtttgg tgacgcttga ggcccaagtc gataccgttg atgccgaaaa
     5701 acaggccacg ttccgagggc gagactggtc gcaagtacgg ccggaatggg ggctagccgg
     5761 caatgctgcc ttcatcgccg caccccgctg gcgcacgcgc gggttggatc ttggcggccg
     5821 cgccttcctg catgattacg attggcgaca cgataaggag tttggcgttt tgaacgtcat
     5881 catgaccgcg cccttgatcg tcgcaaactg gatcaacctg caatactacg gctccacggt
     5941 cgataatctg catcaagggg cgggcaataa agtgctgcat aatgtggtcg gcggcaccgt
     6001 cggggtgatt gaaggcaatg gcggcgacct gcgagttggt cttgccatgc aatcgctgca
     6061 cgatggtgaa cagtggcggc acgaaccgct gcggctctcg gcgtacattg aggcgcccat
     6121 tgcagaaatc gataaaatta ttgccggtca cgacatgctg aacgcgttaa tcaacaaccg
     6181 ctggatgcac atcctgcata tcgatgacaa cggcatcccg catcgccgtc atgcccacgg
     6241 tgactggcga ccggaaccga tctctggttc ttggagtcat ccccagttcg agaaataaat
     6301 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     6361 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     6421 actatatccg gatatccaca ggacgggtgt ggtcgccatg atcgcgtagt cgatagtggc
     6481 tccaagtagc gaagcgagca ggactgggcg gcggccaaag cggtcggaca gtgctccgag
     6541 aacgggtgcg catagaaatt gcatcaacgc atatagcgct agcagcacgc catagtgact
     6601 ggcgatgctg tcggaatgga cgatatcccg caagaggccc ggcagtaccg gcataaccaa
     6661 gcctatgcct acagcatcca gggtgacggt gccgaggatg acgatgagcg cattgttaga
     6721 tttcatacac ggtgcctgac tgcgttagca atttaactgt gataaactac cgcattaaag
     6781 cttatcgatg ataagctgtc aaacatgaga attcttgaag acgaaagggc ctcgtgatac
     6841 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     6901 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     6961 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     7021 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     7081 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     7141 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     7201 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     7261 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     7321 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     7381 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     7441 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     7501 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     7561 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     7621 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     7681 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggctctc
     7741 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     7801 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     7861 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     7921 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     7981 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     8041 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     8101 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     8161 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     8221 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     8281 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     8341 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     8401 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     8461 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     8521 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     8581 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     8641 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     8701 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     8761 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     8821 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
     8881 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     8941 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     9001 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgcctc cgggagctgc
     9061 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     9121 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     9181 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     9241 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     9301 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     9361 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     9421 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     9481 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     9541 tccagacttt acgaaacacg
//
