LOCUS       sequence-262987-        9560 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pET14b carrying the dabA2 gene (Uniprot: D0KWS7) with a c-terminal 
            strep tag fusion 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
FEATURES             Location/Qualifiers
     source          1..9560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     221..240
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     474..491
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    506..645
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(571..590)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     731..753
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(747..938)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2396..2415
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2499..2518
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2499..2517
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2549..2571
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4957..4974
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             7513..7536
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(7588..7606)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7602..7649
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(8015..8020,8021..8037,8038..8043))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(8070..8088)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8156..8260
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8261..8329,8330..9121)
                     /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(8479..8498)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(9292..9560,1..320)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
       61 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
      121 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
      181 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
      241 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
      301 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
      361 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
      421 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
      481 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
      541 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
      601 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
      661 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
      721 gtgaccgcct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
      781 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
      841 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
      901 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
      961 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     1021 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     1081 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     1141 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
     1201 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
     1261 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
     1321 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
     1381 aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg
     1441 cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc
     1501 acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg
     1561 tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg
     1621 ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg
     1681 ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt aggctggtaa
     1741 gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca
     1801 tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg
     1861 ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc
     1921 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     1981 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     2041 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     2101 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     2161 aggagctgac tgggttgaag gctctcaagg gcatcggtcg acgctctccc ttatgcgact
     2221 cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga
     2281 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     2341 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     2401 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     2461 gtccggcgta gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggac
     2521 accacaacgg tttccctcta gaaataattt tgtttaactt taagaaggag atataccatg
     2581 tccatgacta tagagttcag ccacacaacc ggtcttctcc tgctggccat gcccgcactg
     2641 ctccttatgg ccgctgtaat caaacccaag caaggtgctg cttatgcgcg cgcttatcgg
     2701 caacgggttc aatggaccag ttttgcggcc tttggcctgg ccctgctcgc tgtcgtcagt
     2761 tttctatttg caaggcaaca gaatctgatg ctcggtgccg gtagcctccc tgccggattg
     2821 ggattattgg cgctttcaat tcaagtcaat ggcctgactc tggtgctggc aagtctggtt
     2881 agttttgtgc ttagcgtgat tgcccgctac agcgtgcagt atctcgatgg tgatccccag
     2941 caggcccgct ttttccgctt gctggcggtc accggcggat ttttcctgtt ggtcgtcata
     3001 tcaggcaatt tgggattatt caccctcgcc atcattgcga ccggttttgg tttgcatcga
     3061 ctgctttctt tttatgccga tcggccgcgc gcgatcatgg ccacacataa aaaatcaatt
     3121 ttcagccgga ctgccgacgc ccttttgctc gcggccaccg tcttgatcgg tcatcaaatc
     3181 ggtagtttgg agttcagcca aatcagcgcc tatgttcatg cgcaagacca cctatccatt
     3241 gcgttgcacg tggctgcttg gctgatcgtg ctggccgcca tccttaaaag cgcgcaattt
     3301 ccgtttcatg gctggctgat tcaggtcatg gaagcaccca cgccggtttc cgcgctcatg
     3361 cacgccggtg tcgtttacag cggggcaatc attgtgctgc gcaccagcga attattggcc
     3421 gccgacggta ccgccctgtt gcttctggcc ctgattggtt tgatgacctt ggctatcggc
     3481 tcgttggtca tgctgaccca atcggcgatc aaatcatcac tggcctggtc taccgccgct
     3541 cagctcggct tcatgatgct cgaactgggt ttggggttgt ttggcttggc cttgctgcac
     3601 ttggttggcc actcgctcta caaggcccat gcgttcttgt cctcgggcag catgaccgat
     3661 cacctgcgtc aggccaaagt gctcaaaaac cgcccaatct ctgttgtggc ctggtttacc
     3721 acggtgattg tgagcggctt gttcaccctc ggtatcgcgg cggcgatggg tttatcgatc
     3781 gatcaggagc cgatgttgcc cgccgtgctg accatcatcg ccttggcgac cgctcagttg
     3841 atgctcaagg cattgagcca tggcacatgg cgtgaaattc tggtcgccgc aggcgcggca
     3901 atcgccatga ccggcgtgta tgtcttcctg catgaggtgt tcatcacagg ctttgccgac
     3961 acgctggccg cgacaccgca acgtgcgccg ctgctcgatc tgctgttgat ggcgatcacg
     4021 atcataacct tcctgtttgt tgcctggctt caaggccccg gcaaaacctt gatgagcccg
     4081 gagcgacaat tcgccttgtt tgttcacctc aataacgggc tgtatctcga ccgctgggtc
     4141 gaacgcttgg ccttccgttt ctggccggag aaagttggcc gcgcaccgaa aaaatcctgc
     4201 gccgtgattc cgcctaaccc atccggtatc gagccgtctg gttcttctaa aggtgaagaa
     4261 ctgttcaccg gtgttgttcc gatcctggtt gaactggatg gtgatgttaa cggccacaaa
     4321 ttctctgttc gtggtgaagg tgaaggtgat gcaaccaacg gtaaactgac cctgaaattc
     4381 atctgcacta ccggtaaact gccggttcca tggccgactc tggtgactac cctgacctat
     4441 ggtgttcagt gtttttctcg ttacccggat cacatgaagc agcatgattt cttcaaatct
     4501 gcaatgccgg aaggttatgt acaggagcgc accatttctt tcaaagacga tggcacctac
     4561 aaaacccgtg cagaggttaa atttgaaggt gatactctgg tgaaccgtat tgaactgaaa
     4621 ggcattgatt tcaaagagga cggcaacatc ctgggccaca aactggaata taacttcaac
     4681 tcccataacg tttacatcac cgcagacaaa cagaagaacg gtatcaaagc taacttcaaa
     4741 attcgccata acgttgaaga cggtagcgta cagctggcgg accactacca gcagaacact
     4801 ccgatcggtg atggtccggt tctgctgccg gataaccact acctgtccac ccagtctaaa
     4861 ctgtccaaag acccgaacga aaagcgcgac cacatggtgc tgctggagtt cgttactgca
     4921 gcaggtatca cgcacggcat ggatgagctc tacaaacacc atcatcatca tcattaaatc
     4981 cctgccacaa aggcacacac catttacccg aggatttcga ccatgacgac cctgacatca
     5041 ttacaacgca gcgaagcgca acgaaaccac attgttgacc tgattgacaa agcctgcctg
     5101 cgcatcgcgc ccatctggcc actggattcg tttgtcgccg taaaccccta tcttggtttg
     5161 attgatcaac cctttgatac ggtgggtcgg tatctggaac aaaccgtggg cgaatcccta
     5221 tttatggatc acggctggtt tgccgacaag atcgcccaag gggaaatcac cgatgacgat
     5281 ttggcgcagg cggcgcagca actggaccca tcaattagcc ttgataccat caagcagcaa
     5341 ctggccgtcc atcgacaacc tgcacccgcg ctgcctttgg tcacaaacga gctcgatcgc
     5401 cgcgacgcgc cgccagtatc tgaatttgtc atcgagcaag tcagccagtt catggccaac
     5461 tactatgatc gcgggcaagc cctctggcac ttacccaaag aagccagtgc cagtttattc
     5521 gcccaatggc gccgctacac cttgattgac cgcagcgcca gcgccgtcgg cttaaagcag
     5581 gttcgccaac acttgcttgc cgtcccaagc gatgccattg atgcattgtt ctgggcgctg
     5641 gaccagatca atctgcccga aagtcgcctg cccgactacc tgttcacctt attaaaaacc
     5701 atcggtggct gggcgagctg gtgccgctat ctacactttc aagccggttt gcacggcgag
     5761 tcgcaacacg atttacgcga tttgctgatc attcgcttgg tctgggttgc tttggtgatc
     5821 aaagaaacca gctcggctgg acggcaacaa tggcgcgcga agctaaacga ttggtttgat
     5881 ccggccaaat tggtcgcaag cccttcggcg acagctactg cttcgactaa ggctcaatcc
     5941 agccggatcg atgaaatcct gctggcggcc gccgagcagg cgtttcgacg ccggatcaat
     6001 gccggcttga accgccagcc tgccgatgca cccgatcagc aagccgaacg cccgactgtt
     6061 caggccgcgt tttgcattga cgtgcgttcc gaggtgttcc gccgccacct cgaagccagc
     6121 agcccggggc tcgaaaccat cggttttgcc ggttttttcg gcctgccgat tgattattgc
     6181 cgcatgggcg aatcagaggc tcggctacaa aacccggtgc tcatcaaccc cgcctaccgt
     6241 gcgcaggaga cgggcgatcc ggccattgcc caacatcgcc atgcccgaca aagccgaggg
     6301 gccatctgga agcaattcaa actttccgcc gcctcgtgct tcacctttgt cgaatccgca
     6361 ggcttgagct acgtgccgcg tttgctggcc gattcactgg gttggcatcg ctcatcgctg
     6421 ccgcctgatg cccccgggct tacccctgag gagcgtgcgc gtttgcatcc ccaattggtc
     6481 aaactcgatg gcggcgcact atcgacgcag gaaaaagtcg atctggccga aaaagtactc
     6541 cgcgggctgg ggttaacgca cacctttgcg cccatcgtcc tgctcgccgg acacggcagc
     6601 tccaccacca acaacccgca tcgcgcgggg ttggattgcg gtgcctgcgc cggtcaggca
     6661 ggcgatgtga atgctcgcgt cgcggtgcag ttgctcaatg aggccgccgt gcgactgggg
     6721 ctgatcgaac ggggtattgc catcccgcgt gacacccggt tcgtggccgc cctgcacgac
     6781 accaccacag accacatcga actactggat ttggatcaat caggcatcga gagtgatcaa
     6841 ttgtcatcct tgacccaggc gctcaaacaa gccggtgagc tcactcgtct ggagcgtttg
     6901 gtgacgcttg aggcccaagt cgataccgtt gatgccgaaa aacaggccac gttccgaggg
     6961 cgagactggt cgcaagtacg gccggaatgg gggctagccg gcaatgctgc cttcatcgcc
     7021 gcaccccgct ggcgcacgcg cgggttggat cttggcggcc gcgccttcct gcatgattac
     7081 gattggcgac acgataagga gtttggcgtt ttgaacgtca tcatgaccgc gcccttgatc
     7141 gtcgcaaact ggatcaacct gcaatactac ggctccacgg tcgataatct gcatcaaggg
     7201 gcgggcaata aagtgctgca taatgtggtc ggcggcaccg tcggggtgat tgaaggcaat
     7261 ggcggcgacc tgcgagttgg tcttgccatg caatcgctgc acgatggtga acagtggcgg
     7321 cacgaaccgc tgcggctctc ggcgtacatt gaggcgccca ttgcagaaat cgataaaatt
     7381 attgccggtc acgacatgct gaacgcgtta atcaacaacc gctggatgca catcctgcat
     7441 atcgatgaca acggcatccc gcatcgccgt catgcccacg gtgactggcg accggaaccg
     7501 atctctggtt cttggagtca tccccagttc gagaaataaa tccggctgct aacaaagccc
     7561 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     7621 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatatccac
     7681 aggacgggtg tggtcgccat gatcgcgtag tcgatagtgg ctccaagtag cgaagcgagc
     7741 aggactgggc ggcggccaaa gcggtcggac agtgctccga gaacgggtgc gcatagaaat
     7801 tgcatcaacg catatagcgc tagcagcacg ccatagtgac tggcgatgct gtcggaatgg
     7861 acgatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc
     7921 agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga
     7981 ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat gataagctgt
     8041 caaacatgag aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     8101 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     8161 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     8221 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     8281 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     8341 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     8401 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     8461 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     8521 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     8581 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     8641 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     8701 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     8761 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     8821 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     8881 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     8941 gtttattgct gataaatctg gagccggtga gcgtggctct cgcggtatca ttgcagcact
     9001 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     9061 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     9121 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     9181 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     9241 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     9301 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     9361 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     9421 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     9481 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     9541 cgataagtcg tgtcttaccg
//
