LOCUS       sequence-263056-        7317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFE carrying a dabA2 gene with a C351A 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 C351A
FEATURES             Location/Qualifiers
     source          1..7317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     189..208
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        464..568
                     /gene="bla"
                     /label=AmpR promoter
     CDS             569..1192
                     /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(642..661)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      complement(1365..1451)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             5698..5709
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(join(5719..5744,5745..5750,5751..5767,5768..5773,
                     5774..5792))
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    5749..5767
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5774..5792
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(5813..5831)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             6010..6804
                     /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(6064..6083)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6674..6693
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      6835..6929
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      join(7017..7317,1..288)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
       61 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
      121 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
      181 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
      241 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
      301 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
      361 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
      421 cgcagccgaa cgccctaggt ctagcgcgaa ttgtcgaggg aaacgcggaa cccctatttg
      481 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      541 gcttcaataa tattgaaaaa ggaagagtat gtctagatta gataaaagta aagtgattaa
      601 cagcgcatta gagctgctta atgaggtcgg aatcgaaggt ttaacaaccc gtaaactcgc
      661 ccagaagcta ggtgtagagc agcctacatt gtattggcat gtaaaaaata agcgggcttt
      721 gctcgacgcc ttagccattg agatgttaga taggcaccat actcactttt gccctttaga
      781 aggggaaagc tggcaagatt ttttacgtaa taacgctaaa agttttagat gtgctttact
      841 aagtcatcgc gatggagcaa aagtacattt aggtacacgg cctacagaaa aacagtatga
      901 aactctcgaa aatcaattag cctttttatg ccaacaaggt ttttcactag agaatgcatt
      961 atatgcactc agcgctgtgg ggcattttac tttaggttgc gtattggaag atcaagagca
     1021 tcaagtcgct aaagaagaaa gggaaacacc tactactgat agtatgccgc cattattacg
     1081 acaagctatc gaattatttg atcaccaagg tgcagagcca gccttcttat tcggccttga
     1141 attgatcata tgcggattag aaaaacaact taaatgtgaa agtgggtctt aacagttcct
     1201 ggatatccgg atgaaggcac gaacccagtg gacataaccc tgataaatgc ttcaataata
     1261 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttgctt
     1321 ggattctgcg tttgtttccg tctacgaact gctagcggcg gcggatttgt cctactcagg
     1381 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt
     1441 cgttttattt gatgcctcta gcacgcgtac catgggatcc cccgggctgc aggaattccg
     1501 cttcagatcg gttccggtcg ccagtcaccg tgggcatgac ggcgatgcgg gatgccgttg
     1561 tcatcgatat gcaggatgtg catccagcgg ttgttgatta acgcgttcag catgtcgtga
     1621 ccggcaataa ttttatcgat ttctgcaatg ggcgcctcaa tgtacgccga gagccgcagc
     1681 ggttcgtgcc gccactgttc accatcgtgc agcgattgca tggcaagacc aactcgcagg
     1741 tcgccgccat tgccttcaat caccccgacg gtgccgccga ccacattatg cagcacttta
     1801 ttgcccgccc cttgatgcag attatcgacc gtggagccgt agtattgcag gttgatccag
     1861 tttgcgacga tcaagggcgc ggtcatgatg acgttcaaaa cgccaaactc cttatcgtgt
     1921 cgccaatcgt aatcatgcag gaaggcgcgg ccgccaagat ccaacccgcg cgtgcgccag
     1981 cggggtgcgg cgatgaaggc agcattgccg gctagccccc attccggccg tacttgcgac
     2041 cagtctcgcc ctcggaacgt ggcctgtttt tcggcatcaa cggtatcgac ttgggcctca
     2101 agcgtcacca aacgctccag acgagtgagc tcaccggctt gtttgagcgc ctgggtcaag
     2161 gatgacaatt gatcactctc gatgcctgat tgatccaaat ccagtagttc gatgtggtct
     2221 gtggtggtgt cgtgcagggc ggccacgaac cgggtgtcac gcgggatggc aataccccgt
     2281 tcgatcagcc ccagtcgcac ggcggcctca ttgagcaact gcaccgcgac gcgagcattc
     2341 acatcgcctg cctgaccggc gcaggcaccg caatccaacc ccgcgcgatg cgggttgttg
     2401 gtggtggagc tgccgtgtcc ggcgagcagg acgatgggcg caaaggtgtg cgttaacccc
     2461 agcccgcgga gtactttttc ggccagatcg actttttcct gcgtcgatag tgcgccgcca
     2521 tcgagtttga ccaattgggg atgcaaacgc gcacgctcct caggggtaag cccgggggca
     2581 tcaggcggca gcgatgagcg atgccaaccc agtgaatcgg ccagcaaacg cggcacgtag
     2641 ctcaagcctg cggattcgac aaaggtgaag cacgaggcgg cggaaagttt gaattgcttc
     2701 cagatggccc ctcggctttg tcgggcatgg cgatgttggg caatggccgg atcgcccgtc
     2761 tcctgcgcac ggtaggcggg gttgatgagc accgggtttt gtagccgagc ctctgattcg
     2821 cccatgcggc aataatcaat cggcaggccg aaaaaaccgg caaaaccgat ggtttcgagc
     2881 cccgggctgc tggcttcgag gtggcggcgg aacacctcgg aacgcacgtc aatggcaaac
     2941 gcggcctgaa cagtcgggcg ttcggcttgc tgatcgggtg catcggcagg ctggcggttc
     3001 aagccggcat tgatccggcg tcgaaacgcc tgctcggcgg ccgccagcag gatttcatcg
     3061 atccggctgg attgagcctt agtcgaagca gtagctgtcg ccgaagggct tgcgaccaat
     3121 ttggccggat caaaccaatc gtttagcttc gcgcgccatt gttgccgtcc agccgagctg
     3181 gtttctttga tcaccaaagc aacccagacc aagcgaatga tcagcaaatc gcgtaaatcg
     3241 tgttgcgact cgccgtgcaa accggcttga aagtgtagat agcggcacca gctcgcccag
     3301 ccaccgatgg tttttaataa ggtgaacagg tagtcgggca ggcgactttc gggcagattg
     3361 atctggtcca gcgcccagaa caatgcatca atggcatcgc ttgggacggc aagcaagtgt
     3421 tggcgaacct gctttaagcc gacggcgctg gcgctgcggt caatcaaggt gtagcggcgc
     3481 cattgggcga ataaactggc actggcttct ttgggtaagt gccagagggc ttgcccgcga
     3541 tcatagtagt tggccatgaa ctggctgact tgctcgatga caaattcaga tactggcggc
     3601 gcgtcgcggc gatcgagctc gtttgtgacc aaaggcagcg cgggtgcagg ttgtcgatgg
     3661 acggccagtt gctgcttgat ggtatcaagg ctaattgatg ggtccagttg ctgcgccgcc
     3721 tgcgccaaat cgtcatcggt gatttcccct tgggcgatct tgtcggcaaa ccagccgtga
     3781 tccataaata gggattcgcc cacggtttgt tccagatacc gacccaccgt atcaaagggt
     3841 tgatcaatca aaccaagata ggggtttacg gcgacaaacg aatccagtgg ccagatgggc
     3901 gcgatgcgca ggcaggcttt gtcaatcagg tcaacaatgt ggtttcgttg cgcttcgctg
     3961 cgttgtaatg atgtcagggt cgtcatggtc gaaatcctcg ggtaaatggt gtgtgccttt
     4021 gtggcaggga tttacggctc gataccggat gggttaggcg gaatcacggc gcaggatttt
     4081 ttcggtgcgc ggccaacttt ctccggccag aaacggaagg ccaagcgttc gacccagcgg
     4141 tcgagataca gcccgttatt gaggtgaaca aacaaggcga attgtcgctc cgggctcatc
     4201 aaggttttgc cggggccttg aagccaggca acaaacagga aggttatgat cgtgatcgcc
     4261 atcaacagca gatcgagcag cggcgcacgt tgcggtgtcg cggccagcgt gtcggcaaag
     4321 cctgtgatga acacctcatg caggaagaca tacacgccgg tcatggcgat tgccgcgcct
     4381 gcggcgacca gaatttcacg ccatgtgcca tggctcaatg ccttgagcat caactgagcg
     4441 gtcgccaagg cgatgatggt cagcacggcg ggcaacatcg gctcctgatc gatcgataaa
     4501 cccatcgccg ccgcgatacc gagggtgaac aagccgctca caatcaccgt ggtaaaccag
     4561 gccacaacag agattgggcg gtttttgagc actttggcct gacgcaggtg atcggtcatg
     4621 ctgcccgagg acaagaacgc atgggccttg tagagcgagt ggccaaccaa gtgcagcaag
     4681 gccaagccaa acaaccccaa acccagttcg agcatcatga agccgagctg agcggcggta
     4741 gaccaggcca gtgatgattt gatcgccgat tgggtcagca tgaccaacga gccgatagcc
     4801 aaggtcatca aaccaatcag ggccagaagc aacagggcgg taccgtcggc ggccaataat
     4861 tcgctggtgc gcagcacaat gattgccccg ctgtaaacga caccggcgtg catgagcgcg
     4921 gaaaccggcg tgggtgcttc catgacctga atcagccagc catgaaacgg aaattgcgcg
     4981 cttttaagga tggcggccag cacgatcagc caagcagcca cgtgcaacgc aatggatagg
     5041 tggtcttgcg catgaacata ggcgctgatt tggctgaact ccaaactacc gatttgatga
     5101 ccgatcaaga cggtggccgc gagcaaaagg gcgtcggcag tccggctgaa aattgatttt
     5161 ttatgtgtgg ccatgatcgc gcgcggccga tcggcataaa aagaaagcag tcgatgcaaa
     5221 ccaaaaccgg tcgcaatgat ggcgagggtg aataatccca aattgcctga tatgacgacc
     5281 aacaggaaaa atccgccggt gaccgccagc aagcggaaaa agcgggcctg ctggggatca
     5341 ccatcgagat actgcacgct gtagcgggca atcacgctaa gcacaaaact aaccagactt
     5401 gccagcacca gagtcaggcc attgacttga attgaaagcg ccaataatcc caatccggca
     5461 gggaggctac cggcaccgag catcagattc tgttgccttg caaatagaaa actgacgaca
     5521 gcgagcaggg ccaggccaaa ggccgcaaaa ctggtccatt gaacccgttg ccgataagcg
     5581 cgcgcataag cagcaccttg cttgggtttg attacagcgg ccataaggag cagtgcgggc
     5641 atggccagca ggagaagacc ggttgtgtgg ctgaactcta tagtcatgga cggtaccttt
     5701 ctcctcttta atgaattcgg tcagtgcgtc ctgctgatgt gctcagtatc tctatcactg
     5761 atagggatgt caatctctat cactgatagg gactcgaggt gaagacgaaa gggcctcgtg
     5821 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcggaattg
     5881 ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc
     5941 ttgccgccaa ggatctgatg gcgcagggga tcaagatctg atcaagagac aggatgagga
     6001 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     6061 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     6121 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     6181 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     6241 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     6301 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     6361 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     6421 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     6481 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     6541 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     6601 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     6661 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     6721 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     6781 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgagagctcg cttggactcc
     6841 tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga acgctcggtt
     6901 gccgccgggc gttttttatt ggtgagaatc caagcactag tcatgaccaa aatcccttaa
     6961 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7021 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7081 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7141 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     7201 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7261 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataag
//
