LOCUS       sequence-263093-        7317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFE carrying a dabA2 gene (Uniprot: D0KWS7) with a C539A 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 C539A
FEATURES             Location/Qualifiers
     source          1..7317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             953..964
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(join(974..999,1000..1005,1006..1022,1023..1028,
                     1029..1047))
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1004..1022
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1029..1047
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(1068..1086)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             1265..2059
                     /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(1319..1338)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1929..1948
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      2090..2184
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2272..2860
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2761..2780
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3036..3140
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3141..3764
                     /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(3214..3233)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      complement(3937..4023)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggccagtgat gatttgatcg ccgattgggt cagcatgacc aacgagccga tagccaaggt
       61 catcaaacca atcagggcca gaagcaacag ggcggtaccg tcggcggcca ataattcgct
      121 ggtgcgcagc acaatgattg ccccgctgta aacgacaccg gcgtgcatga gcgcggaaac
      181 cggcgtgggt gcttccatga cctgaatcag ccagccatga aacggaaatt gcgcgctttt
      241 aaggatggcg gccagcacga tcagccaagc agccacgtgc aacgcaatgg ataggtggtc
      301 ttgcgcatga acataggcgc tgatttggct gaactccaaa ctaccgattt gatgaccgat
      361 caagacggtg gccgcgagca aaagggcgtc ggcagtccgg ctgaaaattg attttttatg
      421 tgtggccatg atcgcgcgcg gccgatcggc ataaaaagaa agcagtcgat gcaaaccaaa
      481 accggtcgca atgatggcga gggtgaataa tcccaaattg cctgatatga cgaccaacag
      541 gaaaaatccg ccggtgaccg ccagcaagcg gaaaaagcgg gcctgctggg gatcaccatc
      601 gagatactgc acgctgtagc gggcaatcac gctaagcaca aaactaacca gacttgccag
      661 caccagagtc aggccattga cttgaattga aagcgccaat aatcccaatc cggcagggag
      721 gctaccggca ccgagcatca gattctgttg ccttgcaaat agaaaactga cgacagcgag
      781 cagggccagg ccaaaggccg caaaactggt ccattgaacc cgttgccgat aagcgcgcgc
      841 ataagcagca ccttgcttgg gtttgattac agcggccata aggagcagtg cgggcatggc
      901 cagcaggaga agaccggttg tgtggctgaa ctctatagtc atggacggta cctttctcct
      961 ctttaatgaa ttcggtcagt gcgtcctgct gatgtgctca gtatctctat cactgatagg
     1021 gatgtcaatc tctatcactg atagggactc gaggtgaaga cgaaagggcc tcgtgatacg
     1081 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcgg aattgccagc
     1141 tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc
     1201 gccaaggatc tgatggcgca ggggatcaag atctgatcaa gagacaggat gaggatcgtt
     1261 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     1321 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     1381 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     1441 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     1501 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     1561 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     1621 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     1681 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     1741 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     1801 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     1861 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     1921 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     1981 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     2041 tcttgacgag ttcttctgag cgggactctg gggttcgaga gctcgcttgg actcctgttg
     2101 atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc
     2161 cgggcgtttt ttattggtga gaatccaagc actagtcatg accaaaatcc cttaacgtga
     2221 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2281 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2341 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2401 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2461 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2521 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     2581 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2641 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2701 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2761 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2821 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2881 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     2941 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     3001 aacgccctag gtctagcgcg aattgtcgag ggaaacgcgg aacccctatt tgtttatttt
     3061 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3121 aatattgaaa aaggaagagt atgtctagat tagataaaag taaagtgatt aacagcgcat
     3181 tagagctgct taatgaggtc ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc
     3241 taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg
     3301 ccttagccat tgagatgtta gataggcacc atactcactt ttgcccttta gaaggggaaa
     3361 gctggcaaga ttttttacgt aataacgcta aaagttttag atgtgcttta ctaagtcatc
     3421 gcgatggagc aaaagtacat ttaggtacac ggcctacaga aaaacagtat gaaactctcg
     3481 aaaatcaatt agccttttta tgccaacaag gtttttcact agagaatgca ttatatgcac
     3541 tcagcgctgt ggggcatttt actttaggtt gcgtattgga agatcaagag catcaagtcg
     3601 ctaaagaaga aagggaaaca cctactactg atagtatgcc gccattatta cgacaagcta
     3661 tcgaattatt tgatcaccaa ggtgcagagc cagccttctt attcggcctt gaattgatca
     3721 tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc ttaacagttc ctggatatcc
     3781 ggatgaaggc acgaacccag tggacataac cctgataaat gcttcaataa tattgaaaaa
     3841 ggaagagtat gagtattcaa catttccgtg tcgcccttat tcccttttgc ttggattctg
     3901 cgtttgtttc cgtctacgaa ctgctagcgg cggcggattt gtcctactca ggagagcgtt
     3961 caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat
     4021 ttgatgcctc tagcacgcgt accatgggat cccccgggct gcaggaattc cgcttcagat
     4081 cggttccggt cgccagtcac cgtgggcatg acggcgatgc gggatgccgt tgtcatcgat
     4141 atgcaggatg tgcatccagc ggttgttgat taacgcgttc agcatgtcgt gaccggcaat
     4201 aattttatcg atttctgcaa tgggcgcctc aatgtacgcc gagagccgca gcggttcgtg
     4261 ccgccactgt tcaccatcgt gcagcgattg catggcaaga ccaactcgca ggtcgccgcc
     4321 attgccttca atcaccccga cggtgccgcc gaccacatta tgcagcactt tattgcccgc
     4381 cccttgatgc agattatcga ccgtggagcc gtagtattgc aggttgatcc agtttgcgac
     4441 gatcaagggc gcggtcatga tgacgttcaa aacgccaaac tccttatcgt gtcgccaatc
     4501 gtaatcatgc aggaaggcgc ggccgccaag atccaacccg cgcgtgcgcc agcggggtgc
     4561 ggcgatgaag gcagcattgc cggctagccc ccattccggc cgtacttgcg accagtctcg
     4621 ccctcggaac gtggcctgtt tttcggcatc aacggtatcg acttgggcct caagcgtcac
     4681 caaacgctcc agacgagtga gctcaccggc ttgtttgagc gcctgggtca aggatgacaa
     4741 ttgatcactc tcgatgcctg attgatccaa atccagtagt tcgatgtggt ctgtggtggt
     4801 gtcgtgcagg gcggccacga accgggtgtc acgcgggatg gcaatacccc gttcgatcag
     4861 ccccagtcgc acggcggcct cattgagcaa ctgcaccgcg acgcgagcat tcacatcgcc
     4921 tgcctgaccg gcgcaggcac ccgcatccaa ccccgcgcga tgcgggttgt tggtggtgga
     4981 gctgccgtgt ccggcgagca ggacgatggg cgcaaaggtg tgcgttaacc ccagcccgcg
     5041 gagtactttt tcggccagat cgactttttc ctgcgtcgat agtgcgccgc catcgagttt
     5101 gaccaattgg ggatgcaaac gcgcacgctc ctcaggggta agcccggggg catcaggcgg
     5161 cagcgatgag cgatgccaac ccagtgaatc ggccagcaaa cgcggcacgt agctcaagcc
     5221 tgcggattcg acaaaggtga agcacgaggc ggcggaaagt ttgaattgct tccagatggc
     5281 ccctcggctt tgtcgggcat ggcgatgttg ggcaatggcc ggatcgcccg tctcctgcgc
     5341 acggtaggcg gggttgatga gcaccgggtt ttgtagccga gcctctgatt cgcccatgcg
     5401 gcaataatca atcggcaggc cgaaaaaacc ggcaaaaccg atggtttcga gccccgggct
     5461 gctggcttcg aggtggcggc ggaacacctc ggaacgcacg tcaatgcaaa acgcggcctg
     5521 aacagtcggg cgttcggctt gctgatcggg tgcatcggca ggctggcggt tcaagccggc
     5581 attgatccgg cgtcgaaacg cctgctcggc ggccgccagc aggatttcat cgatccggct
     5641 ggattgagcc ttagtcgaag cagtagctgt cgccgaaggg cttgcgacca atttggccgg
     5701 atcaaaccaa tcgtttagct tcgcgcgcca ttgttgccgt ccagccgagc tggtttcttt
     5761 gatcaccaaa gcaacccaga ccaagcgaat gatcagcaaa tcgcgtaaat cgtgttgcga
     5821 ctcgccgtgc aaaccggctt gaaagtgtag atagcggcac cagctcgccc agccaccgat
     5881 ggtttttaat aaggtgaaca ggtagtcggg caggcgactt tcgggcagat tgatctggtc
     5941 cagcgcccag aacaatgcat caatggcatc gcttgggacg gcaagcaagt gttggcgaac
     6001 ctgctttaag ccgacggcgc tggcgctgcg gtcaatcaag gtgtagcggc gccattgggc
     6061 gaataaactg gcactggctt ctttgggtaa gtgccagagg gcttgcccgc gatcatagta
     6121 gttggccatg aactggctga cttgctcgat gacaaattca gatactggcg gcgcgtcgcg
     6181 gcgatcgagc tcgtttgtga ccaaaggcag cgcgggtgca ggttgtcgat ggacggccag
     6241 ttgctgcttg atggtatcaa ggctaattga tgggtccagt tgctgcgccg cctgcgccaa
     6301 atcgtcatcg gtgatttccc cttgggcgat cttgtcggca aaccagccgt gatccataaa
     6361 tagggattcg cccacggttt gttccagata ccgacccacc gtatcaaagg gttgatcaat
     6421 caaaccaaga taggggttta cggcgacaaa cgaatccagt ggccagatgg gcgcgatgcg
     6481 caggcaggct ttgtcaatca ggtcaacaat gtggtttcgt tgcgcttcgc tgcgttgtaa
     6541 tgatgtcagg gtcgtcatgg tcgaaatcct cgggtaaatg gtgtgtgcct ttgtggcagg
     6601 gatttacggc tcgataccgg atgggttagg cggaatcacg gcgcaggatt ttttcggtgc
     6661 gcggccaact ttctccggcc agaaacggaa ggccaagcgt tcgacccagc ggtcgagata
     6721 cagcccgtta ttgaggtgaa caaacaaggc gaattgtcgc tccgggctca tcaaggtttt
     6781 gccggggcct tgaagccagg caacaaacag gaaggttatg atcgtgatcg ccatcaacag
     6841 cagatcgagc agcggcgcac gttgcggtgt cgcggccagc gtgtcggcaa agcctgtgat
     6901 gaacacctca tgcaggaaga catacacgcc ggtcatggcg attgccgcgc ctgcggcgac
     6961 cagaatttca cgccatgtgc catggctcaa tgccttgagc atcaactgag cggtcgccaa
     7021 ggcgatgatg gtcagcacgg cgggcaacat cggctcctga tcgatcgata aacccatcgc
     7081 cgccgcgata ccgagggtga acaagccgct cacaatcacc gtggtaaacc aggccacaac
     7141 agagattggg cggtttttga gcactttggc ctgacgcagg tgatcggtca tgctgcccga
     7201 ggacaagaac gcatgggcct tgtagagcga gtggccaacc aagtgcagca aggccaagcc
     7261 aaacaacccc aaacccagtt cgagcatcat gaagccgagc tgagcggcgg tagacca
//
