LOCUS       sequence-263046-        7317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFE carrying a dabA2 gene (Uniprot: D0KWS7) with a H524A mutation 
            and a dabB2 gene (Uniprot: D0KWS8).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7317)
  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 7317)
  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: pFE-dabAB2 H524A
FEATURES             Location/Qualifiers
     source          1..7317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      84..672
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     573..592
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        848..952
                     /gene="bla"
                     /label=AmpR promoter
     CDS             953..1576
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(1026..1045)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      complement(1749..1835)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             6082..6093
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(join(6103..6128,6129..6134,6135..6151,6152..6157,
                     6158..6176))
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    6133..6151
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6158..6176
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(6197..6215)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             6394..7188
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6448..6467)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7058..7077
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      7219..7313
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 gcactagtca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
       61 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
      121 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
      181 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
      241 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
      301 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
      361 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
      421 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
      481 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
      541 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
      601 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
      661 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
      721 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
      781 tttgagtgag ctgataccgc tcgccgcagc cgaacgccct aggtctagcg cgaattgtcg
      841 agggaaacgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
      901 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgtctag
      961 attagataaa agtaaagtga ttaacagcgc attagagctg cttaatgagg tcggaatcga
     1021 aggtttaaca acccgtaaac tcgcccagaa gctaggtgta gagcagccta cattgtattg
     1081 gcatgtaaaa aataagcggg ctttgctcga cgccttagcc attgagatgt tagataggca
     1141 ccatactcac ttttgccctt tagaagggga aagctggcaa gattttttac gtaataacgc
     1201 taaaagtttt agatgtgctt tactaagtca tcgcgatgga gcaaaagtac atttaggtac
     1261 acggcctaca gaaaaacagt atgaaactct cgaaaatcaa ttagcctttt tatgccaaca
     1321 aggtttttca ctagagaatg cattatatgc actcagcgct gtggggcatt ttactttagg
     1381 ttgcgtattg gaagatcaag agcatcaagt cgctaaagaa gaaagggaaa cacctactac
     1441 tgatagtatg ccgccattat tacgacaagc tatcgaatta tttgatcacc aaggtgcaga
     1501 gccagccttc ttattcggcc ttgaattgat catatgcgga ttagaaaaac aacttaaatg
     1561 tgaaagtggg tcttaacagt tcctggatat ccggatgaag gcacgaaccc agtggacata
     1621 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     1681 tgtcgccctt attccctttt gcttggattc tgcgtttgtt tccgtctacg aactgctagc
     1741 ggcggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag
     1801 gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tctagcacgc gtaccatggg
     1861 atcccccggg ctgcaggaat tccgcttcag atcggttccg gtcgccagtc accgtgggca
     1921 tgacggcgat gcgggatgcc gttgtcatcg atatgcagga tgtgcatcca gcggttgttg
     1981 attaacgcgt tcagcatgtc gtgaccggca ataattttat cgatttctgc aatgggcgcc
     2041 tcaatgtacg ccgagagccg cagcggttcg tgccgccact gttcaccatc gtgcagcgat
     2101 tgcatggcaa gaccaactcg caggtcgccg ccattgcctt caatcacccc gacggtgccg
     2161 ccgaccacat tatgcagcac tttattgccc gccccttgat gcagattatc gaccgtggag
     2221 ccgtagtatt gcaggttgat ccagtttgcg acgatcaagg gcgcggtcat gatgacgttc
     2281 aaaacgccaa actccttatc gtgtcgccaa tcgtaatcat gcaggaaggc gcggccgcca
     2341 agatccaacc cgcgcgtgcg ccagcggggt gcggcgatga aggcagcatt gccggctagc
     2401 ccccattccg gccgtacttg cgaccagtct cgccctcgga acgtggcctg tttttcggca
     2461 tcaacggtat cgacttgggc ctcaagcgtc accaaacgct ccagacgagt gagctcaccg
     2521 gcttgtttga gcgcctgggt caaggatgac aattgatcac tctcgatgcc tgattgatcc
     2581 aaatccagta gttcgatgtg gtctgtggtg gtgtcgtgca gggcggccac gaaccgggtg
     2641 tcacgcggga tggcaatacc ccgttcgatc agccccagtc gcacggcggc ctcattgagc
     2701 aactgcaccg cgacgcgagc attcacatcg cctgcctgac cggcgcaggc accgcaatcc
     2761 aaccccgcgc gatgcgggtt gttggtggtg gagctgcccg ctccggcgag caggacgatg
     2821 ggcgcaaagg tgtgcgttaa ccccagcccg cggagtactt tttcggccag atcgactttt
     2881 tcctgcgtcg atagtgcgcc gccatcgagt ttgaccaatt ggggatgcaa acgcgcacgc
     2941 tcctcagggg taagcccggg ggcatcaggc ggcagcgatg agcgatgcca acccagtgaa
     3001 tcggccagca aacgcggcac gtagctcaag cctgcggatt cgacaaaggt gaagcacgag
     3061 gcggcggaaa gtttgaattg cttccagatg gcccctcggc tttgtcgggc atggcgatgt
     3121 tgggcaatgg ccggatcgcc cgtctcctgc gcacggtagg cggggttgat gagcaccggg
     3181 ttttgtagcc gagcctctga ttcgcccatg cggcaataat caatcggcag gccgaaaaaa
     3241 ccggcaaaac cgatggtttc gagccccggg ctgctggctt cgaggtggcg gcggaacacc
     3301 tcggaacgca cgtcaatgca aaacgcggcc tgaacagtcg ggcgttcggc ttgctgatcg
     3361 ggtgcatcgg caggctggcg gttcaagccg gcattgatcc ggcgtcgaaa cgcctgctcg
     3421 gcggccgcca gcaggatttc atcgatccgg ctggattgag ccttagtcga agcagtagct
     3481 gtcgccgaag ggcttgcgac caatttggcc ggatcaaacc aatcgtttag cttcgcgcgc
     3541 cattgttgcc gtccagccga gctggtttct ttgatcacca aagcaaccca gaccaagcga
     3601 atgatcagca aatcgcgtaa atcgtgttgc gactcgccgt gcaaaccggc ttgaaagtgt
     3661 agatagcggc accagctcgc ccagccaccg atggttttta ataaggtgaa caggtagtcg
     3721 ggcaggcgac tttcgggcag attgatctgg tccagcgccc agaacaatgc atcaatggca
     3781 tcgcttggga cggcaagcaa gtgttggcga acctgcttta agccgacggc gctggcgctg
     3841 cggtcaatca aggtgtagcg gcgccattgg gcgaataaac tggcactggc ttctttgggt
     3901 aagtgccaga gggcttgccc gcgatcatag tagttggcca tgaactggct gacttgctcg
     3961 atgacaaatt cagatactgg cggcgcgtcg cggcgatcga gctcgtttgt gaccaaaggc
     4021 agcgcgggtg caggttgtcg atggacggcc agttgctgct tgatggtatc aaggctaatt
     4081 gatgggtcca gttgctgcgc cgcctgcgcc aaatcgtcat cggtgatttc cccttgggcg
     4141 atcttgtcgg caaaccagcc gtgatccata aatagggatt cgcccacggt ttgttccaga
     4201 taccgaccca ccgtatcaaa gggttgatca atcaaaccaa gataggggtt tacggcgaca
     4261 aacgaatcca gtggccagat gggcgcgatg cgcaggcagg ctttgtcaat caggtcaaca
     4321 atgtggtttc gttgcgcttc gctgcgttgt aatgatgtca gggtcgtcat ggtcgaaatc
     4381 ctcgggtaaa tggtgtgtgc ctttgtggca gggatttacg gctcgatacc ggatgggtta
     4441 ggcggaatca cggcgcagga ttttttcggt gcgcggccaa ctttctccgg ccagaaacgg
     4501 aaggccaagc gttcgaccca gcggtcgaga tacagcccgt tattgaggtg aacaaacaag
     4561 gcgaattgtc gctccgggct catcaaggtt ttgccggggc cttgaagcca ggcaacaaac
     4621 aggaaggtta tgatcgtgat cgccatcaac agcagatcga gcagcggcgc acgttgcggt
     4681 gtcgcggcca gcgtgtcggc aaagcctgtg atgaacacct catgcaggaa gacatacacg
     4741 ccggtcatgg cgattgccgc gcctgcggcg accagaattt cacgccatgt gccatggctc
     4801 aatgccttga gcatcaactg agcggtcgcc aaggcgatga tggtcagcac ggcgggcaac
     4861 atcggctcct gatcgatcga taaacccatc gccgccgcga taccgagggt gaacaagccg
     4921 ctcacaatca ccgtggtaaa ccaggccaca acagagattg ggcggttttt gagcactttg
     4981 gcctgacgca ggtgatcggt catgctgccc gaggacaaga acgcatgggc cttgtagagc
     5041 gagtggccaa ccaagtgcag caaggccaag ccaaacaacc ccaaacccag ttcgagcatc
     5101 atgaagccga gctgagcggc ggtagaccag gccagtgatg atttgatcgc cgattgggtc
     5161 agcatgacca acgagccgat agccaaggtc atcaaaccaa tcagggccag aagcaacagg
     5221 gcggtaccgt cggcggccaa taattcgctg gtgcgcagca caatgattgc cccgctgtaa
     5281 acgacaccgg cgtgcatgag cgcggaaacc ggcgtgggtg cttccatgac ctgaatcagc
     5341 cagccatgaa acggaaattg cgcgctttta aggatggcgg ccagcacgat cagccaagca
     5401 gccacgtgca acgcaatgga taggtggtct tgcgcatgaa cataggcgct gatttggctg
     5461 aactccaaac taccgatttg atgaccgatc aagacggtgg ccgcgagcaa aagggcgtcg
     5521 gcagtccggc tgaaaattga ttttttatgt gtggccatga tcgcgcgcgg ccgatcggca
     5581 taaaaagaaa gcagtcgatg caaaccaaaa ccggtcgcaa tgatggcgag ggtgaataat
     5641 cccaaattgc ctgatatgac gaccaacagg aaaaatccgc cggtgaccgc cagcaagcgg
     5701 aaaaagcggg cctgctgggg atcaccatcg agatactgca cgctgtagcg ggcaatcacg
     5761 ctaagcacaa aactaaccag acttgccagc accagagtca ggccattgac ttgaattgaa
     5821 agcgccaata atcccaatcc ggcagggagg ctaccggcac cgagcatcag attctgttgc
     5881 cttgcaaata gaaaactgac gacagcgagc agggccaggc caaaggccgc aaaactggtc
     5941 cattgaaccc gttgccgata agcgcgcgca taagcagcac cttgcttggg tttgattaca
     6001 gcggccataa ggagcagtgc gggcatggcc agcaggagaa gaccggttgt gtggctgaac
     6061 tctatagtca tggacggtac ctttctcctc tttaatgaat tcggtcagtg cgtcctgctg
     6121 atgtgctcag tatctctatc actgataggg atgtcaatct ctatcactga tagggactcg
     6181 aggtgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     6241 atggtttctt agacgtcgga attgccagct ggggcgccct ctggtaaggt tgggaagccc
     6301 tgcaaagtaa actggatggc tttcttgccg ccaaggatct gatggcgcag gggatcaaga
     6361 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     6421 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     6481 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     6541 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     6601 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     6661 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     6721 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     6781 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     6841 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     6901 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     6961 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     7021 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     7081 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     7141 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     7201 ggttcgagag ctcgcttgga ctcctgttga tagatccagt aatgacctca gaactccatc
     7261 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaa
//
