LOCUS       sequence-263065-        7317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFE carrying both the dabA2 (Uniprot: D0KWS7) and dabB2 genes 
            (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
FEATURES             Location/Qualifiers
     source          1..7317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             109..903
                     /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(163..182)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     773..792
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      934..1028
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1116..1704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1605..1624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1880..1984
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1985..2608
                     /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(2058..2077)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      complement(2781..2867)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             7114..7125
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(join(7135..7160,7161..7166,7167..7183,7184..7189,
                     7190..7208))
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    7165..7183
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7190..7208
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(7229..7247)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aaggttggga agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg
       61 cgcaggggat caagatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
      121 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
      181 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
      241 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
      301 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
      361 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
      421 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
      481 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
      541 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
      601 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
      661 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
      721 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
      781 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
      841 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
      901 tgagcgggac tctggggttc gagagctcgc ttggactcct gttgatagat ccagtaatga
      961 cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg
     1021 gtgagaatcc aagcactagt catgaccaaa atcccttaac gtgagttttc gttccactga
     1081 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1141 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1201 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1261 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1321 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1381 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     1441 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     1501 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     1561 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     1621 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     1681 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     1741 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     1801 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgcc ctaggtctag
     1861 cgcgaattgt cgagggaaac gcggaacccc tatttgttta tttttctaaa tacattcaaa
     1921 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     1981 gagtatgtct agattagata aaagtaaagt gattaacagc gcattagagc tgcttaatga
     2041 ggtcggaatc gaaggtttaa caacccgtaa actcgcccag aagctaggtg tagagcagcc
     2101 tacattgtat tggcatgtaa aaaataagcg ggctttgctc gacgccttag ccattgagat
     2161 gttagatagg caccatactc acttttgccc tttagaaggg gaaagctggc aagatttttt
     2221 acgtaataac gctaaaagtt ttagatgtgc tttactaagt catcgcgatg gagcaaaagt
     2281 acatttaggt acacggccta cagaaaaaca gtatgaaact ctcgaaaatc aattagcctt
     2341 tttatgccaa caaggttttt cactagagaa tgcattatat gcactcagcg ctgtggggca
     2401 ttttacttta ggttgcgtat tggaagatca agagcatcaa gtcgctaaag aagaaaggga
     2461 aacacctact actgatagta tgccgccatt attacgacaa gctatcgaat tatttgatca
     2521 ccaaggtgca gagccagcct tcttattcgg ccttgaattg atcatatgcg gattagaaaa
     2581 acaacttaaa tgtgaaagtg ggtcttaaca gttcctggat atccggatga aggcacgaac
     2641 ccagtggaca taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     2701 tcaacatttc cgtgtcgccc ttattccctt ttgcttggat tctgcgtttg tttccgtcta
     2761 cgaactgcta gcggcggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
     2821 ataaaacgaa aggcccagtc tttcgactga gcctttcgtt ttatttgatg cctctagcac
     2881 gcgtaccatg ggatcccccg ggctgcagga attccgcttc agatcggttc cggtcgccag
     2941 tcaccgtggg catgacggcg atgcgggatg ccgttgtcat cgatatgcag gatgtgcatc
     3001 cagcggttgt tgattaacgc gttcagcatg tcgtgaccgg caataatttt atcgatttct
     3061 gcaatgggcg cctcaatgta cgccgagagc cgcagcggtt cgtgccgcca ctgttcacca
     3121 tcgtgcagcg attgcatggc aagaccaact cgcaggtcgc cgccattgcc ttcaatcacc
     3181 ccgacggtgc cgccgaccac attatgcagc actttattgc ccgccccttg atgcagatta
     3241 tcgaccgtgg agccgtagta ttgcaggttg atccagtttg cgacgatcaa gggcgcggtc
     3301 atgatgacgt tcaaaacgcc aaactcctta tcgtgtcgcc aatcgtaatc atgcaggaag
     3361 gcgcggccgc caagatccaa cccgcgcgtg cgccagcggg gtgcggcgat gaaggcagca
     3421 ttgccggcta gcccccattc cggccgtact tgcgaccagt ctcgccctcg gaacgtggcc
     3481 tgtttttcgg catcaacggt atcgacttgg gcctcaagcg tcaccaaacg ctccagacga
     3541 gtgagctcac cggcttgttt gagcgcctgg gtcaaggatg acaattgatc actctcgatg
     3601 cctgattgat ccaaatccag tagttcgatg tggtctgtgg tggtgtcgtg cagggcggcc
     3661 acgaaccggg tgtcacgcgg gatggcaata ccccgttcga tcagccccag tcgcacggcg
     3721 gcctcattga gcaactgcac cgcgacgcga gcattcacat cgcctgcctg accggcgcag
     3781 gcaccgcaat ccaaccccgc gcgatgcggg ttgttggtgg tggagctgcc gtgtccggcg
     3841 agcaggacga tgggcgcaaa ggtgtgcgtt aaccccagcc cgcggagtac tttttcggcc
     3901 agatcgactt tttcctgcgt cgatagtgcg ccgccatcga gtttgaccaa ttggggatgc
     3961 aaacgcgcac gctcctcagg ggtaagcccg ggggcatcag gcggcagcga tgagcgatgc
     4021 caacccagtg aatcggccag caaacgcggc acgtagctca agcctgcgga ttcgacaaag
     4081 gtgaagcacg aggcggcgga aagtttgaat tgcttccaga tggcccctcg gctttgtcgg
     4141 gcatggcgat gttgggcaat ggccggatcg cccgtctcct gcgcacggta ggcggggttg
     4201 atgagcaccg ggttttgtag ccgagcctct gattcgccca tgcggcaata atcaatcggc
     4261 aggccgaaaa aaccggcaaa accgatggtt tcgagccccg ggctgctggc ttcgaggtgg
     4321 cggcggaaca cctcggaacg cacgtcaatg caaaacgcgg cctgaacagt cgggcgttcg
     4381 gcttgctgat cgggtgcatc ggcaggctgg cggttcaagc cggcattgat ccggcgtcga
     4441 aacgcctgct cggcggccgc cagcaggatt tcatcgatcc ggctggattg agccttagtc
     4501 gaagcagtag ctgtcgccga agggcttgcg accaatttgg ccggatcaaa ccaatcgttt
     4561 agcttcgcgc gccattgttg ccgtccagcc gagctggttt ctttgatcac caaagcaacc
     4621 cagaccaagc gaatgatcag caaatcgcgt aaatcgtgtt gcgactcgcc gtgcaaaccg
     4681 gcttgaaagt gtagatagcg gcaccagctc gcccagccac cgatggtttt taataaggtg
     4741 aacaggtagt cgggcaggcg actttcgggc agattgatct ggtccagcgc ccagaacaat
     4801 gcatcaatgg catcgcttgg gacggcaagc aagtgttggc gaacctgctt taagccgacg
     4861 gcgctggcgc tgcggtcaat caaggtgtag cggcgccatt gggcgaataa actggcactg
     4921 gcttctttgg gtaagtgcca gagggcttgc ccgcgatcat agtagttggc catgaactgg
     4981 ctgacttgct cgatgacaaa ttcagatact ggcggcgcgt cgcggcgatc gagctcgttt
     5041 gtgaccaaag gcagcgcggg tgcaggttgt cgatggacgg ccagttgctg cttgatggta
     5101 tcaaggctaa ttgatgggtc cagttgctgc gccgcctgcg ccaaatcgtc atcggtgatt
     5161 tccccttggg cgatcttgtc ggcaaaccag ccgtgatcca taaataggga ttcgcccacg
     5221 gtttgttcca gataccgacc caccgtatca aagggttgat caatcaaacc aagatagggg
     5281 tttacggcga caaacgaatc cagtggccag atgggcgcga tgcgcaggca ggctttgtca
     5341 atcaggtcaa caatgtggtt tcgttgcgct tcgctgcgtt gtaatgatgt cagggtcgtc
     5401 atggtcgaaa tcctcgggta aatggtgtgt gcctttgtgg cagggattta cggctcgata
     5461 ccggatgggt taggcggaat cacggcgcag gattttttcg gtgcgcggcc aactttctcc
     5521 ggccagaaac ggaaggccaa gcgttcgacc cagcggtcga gatacagccc gttattgagg
     5581 tgaacaaaca aggcgaattg tcgctccggg ctcatcaagg ttttgccggg gccttgaagc
     5641 caggcaacaa acaggaaggt tatgatcgtg atcgccatca acagcagatc gagcagcggc
     5701 gcacgttgcg gtgtcgcggc cagcgtgtcg gcaaagcctg tgatgaacac ctcatgcagg
     5761 aagacataca cgccggtcat ggcgattgcc gcgcctgcgg cgaccagaat ttcacgccat
     5821 gtgccatggc tcaatgcctt gagcatcaac tgagcggtcg ccaaggcgat gatggtcagc
     5881 acggcgggca acatcggctc ctgatcgatc gataaaccca tcgccgccgc gataccgagg
     5941 gtgaacaagc cgctcacaat caccgtggta aaccaggcca caacagagat tgggcggttt
     6001 ttgagcactt tggcctgacg caggtgatcg gtcatgctgc ccgaggacaa gaacgcatgg
     6061 gccttgtaga gcgagtggcc aaccaagtgc agcaaggcca agccaaacaa ccccaaaccc
     6121 agttcgagca tcatgaagcc gagctgagcg gcggtagacc aggccagtga tgatttgatc
     6181 gccgattggg tcagcatgac caacgagccg atagccaagg tcatcaaacc aatcagggcc
     6241 agaagcaaca gggcggtacc gtcggcggcc aataattcgc tggtgcgcag cacaatgatt
     6301 gccccgctgt aaacgacacc ggcgtgcatg agcgcggaaa ccggcgtggg tgcttccatg
     6361 acctgaatca gccagccatg aaacggaaat tgcgcgcttt taaggatggc ggccagcacg
     6421 atcagccaag cagccacgtg caacgcaatg gataggtggt cttgcgcatg aacataggcg
     6481 ctgatttggc tgaactccaa actaccgatt tgatgaccga tcaagacggt ggccgcgagc
     6541 aaaagggcgt cggcagtccg gctgaaaatt gattttttat gtgtggccat gatcgcgcgc
     6601 ggccgatcgg cataaaaaga aagcagtcga tgcaaaccaa aaccggtcgc aatgatggcg
     6661 agggtgaata atcccaaatt gcctgatatg acgaccaaca ggaaaaatcc gccggtgacc
     6721 gccagcaagc ggaaaaagcg ggcctgctgg ggatcaccat cgagatactg cacgctgtag
     6781 cgggcaatca cgctaagcac aaaactaacc agacttgcca gcaccagagt caggccattg
     6841 acttgaattg aaagcgccaa taatcccaat ccggcaggga ggctaccggc accgagcatc
     6901 agattctgtt gccttgcaaa tagaaaactg acgacagcga gcagggccag gccaaaggcc
     6961 gcaaaactgg tccattgaac ccgttgccga taagcgcgcg cataagcagc accttgcttg
     7021 ggtttgatta cagcggccat aaggagcagt gcgggcatgg ccagcaggag aagaccggtt
     7081 gtgtggctga actctatagt catggacggt acctttctcc tctttaatga attcggtcag
     7141 tgcgtcctgc tgatgtgctc agtatctcta tcactgatag ggatgtcaat ctctatcact
     7201 gatagggact cgaggtgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     7261 gtcatgataa taatggtttc ttagacgtcg gaattgccag ctggggcgcc ctctggt
//
