LOCUS       sequence-262988-        7289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFE carrying the badabA and badabB genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7289)
  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 7289)
  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-baDAB
FEATURES             Location/Qualifiers
     source          1..7289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(316..410)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(441..1235)
                     /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(552..571)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1162..1181
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1414..1432
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1453..1471,1472..1477,1478..1494,1495..1500,
                     1501..1526)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1453..1471
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1478..1496
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1536..1547
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      5766..5852
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(6025..6648)
                     /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     6556..6575
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        complement(6649..6753)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(6929..7289,1..228))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7009..7028)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
       61 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
      121 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
      181 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
      241 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
      301 atgactagtg cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa
      361 caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct
      421 ctcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc
      481 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca
      541 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc
      601 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag
      661 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg
      721 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca
      781 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat
      841 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact
      901 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc
      961 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc
     1021 gtggccagcc acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg
     1081 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca
     1141 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc
     1201 ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct
     1261 tgatcagatc ttgatcccct gcgccatcag atccttggcg gcaagaaagc catccagttt
     1321 actttgcagg gcttcccaac cttaccagag ggcgccccag ctggcaattc cgacgtctaa
     1381 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     1441 cttcacctcg agtccctatc agtgatagag attgacatcc ctatcagtga tagagatact
     1501 gagcacatca gcaggacgca ctgaccgaat tcattaaaga ggagaaaggt accgtccatg
     1561 ttaatttcgc taagttcatc aacactgtta acattatttt ttatcgcgct tagtgcttct
     1621 tggctaagtg gattattgtt tttacatgca aggatgcctt tacgttttgt tcatatccat
     1681 attggtatcg ctgctttgcc ttcattggtt tctttacttg cacttgttaa taacaatgga
     1741 gatagagttg tggggccttg gcatctagat actttagctt ggttaatggc tttcttcgtt
     1801 cttacaattg gtttaatcat tcagcgattt tctgtacgtt acttaatggg agatcgctca
     1861 tatcgaaaat actttgcact ttttacattt actacaggtg tatcttcagt tgcatggtta
     1921 agtgatgatc ttcgcttcat gattatgtgt tggggtgcaa cacttatagg attggttcta
     1981 ctcatcgggc taaataaagg gtggaaagta gttagcgaag caacaaaaat ctctggctat
     2041 ttatttacaa taagttggat cgccttacta tcagctatca tttggctgtt tcaaataact
     2101 ggacagtggc agttaacatc agttgtaacg aatgaaaatg tagcccaatt tggaacattg
     2161 gagaaaacag gaattaactt attgattata gtagctgtga tgattccagc agcacaatgg
     2221 ccttttcaaa gatggttaat tgaatcagct gttgctccaa ctcctgtttc tgctattatg
     2281 catgcgggtt tagtaaatgc tggcggtatc atgttaacta gattttcgcc actttttcat
     2341 gatgatattg cacaaatcat tttactaatt ttctctagta tttctgtctt aataggaaca
     2401 ggaattagtt tagttcaagt tgattataaa cgtcagctag taggatctac tattgcacaa
     2461 atgggattta tgctgattca atgtgcatta ggcgcttatt tagcagctgt tattcattta
     2521 attttacatg gtttatttaa agccacactg tttttacaag ctggttcctc agtacaacgt
     2581 gttgaggtag tgaagcagtc taataaaaag atgtccaatt tatggatgat agtcgggcgt
     2641 gttttagggt tatttatagc aattgctttc tggtttataa cttctggaga ggggtatcaa
     2701 ttagttagtg cgttaatctt aggctggtca ctatactttt cttggaaaca gcttgttgtt
     2761 tttggagagg gaagaatggg acgaattgct ggcttaattg ttttgattgg attttctctc
     2821 atttacttta ccgttcataa ttctctttat aaatggttgc acactgacat gtatcaaagt
     2881 gttcaacctt cagctccagc cgttatcttt gttatatgta tcttactatt tagtagtgtt
     2941 atttgtactt tcgttactcg taatcaatct tctactttat ccgctgtact ttatctttgg
     3001 tcagttcgag taggtgaggc gagacgaaag tcagtagaaa gtcatccaag ctatctgaaa
     3061 catgatgtat caaagggagg taattcgtga tgagcatacc gtcaatatta aggaaagaga
     3121 ctttaaaaaa gaaagataaa aatattgatc tacaagaaaa taatataaat gatttagttg
     3181 tatcggctag ccgagtgatt gcgccacttt ggcctatctc tacatttgca gctcatcacc
     3241 cttggatggg gctggaaaag caatcatttg aacaagttgc aaattggtta aaagaagctc
     3301 gtaatgtgga tatttatcct agtgcttcca tgattcattc ggcaaaggcg aaaggtgaga
     3361 ttgaggaatc atttttacaa attgccttgt ctcgttggct tgattcacaa tcttttcata
     3421 tgccgcgaga gacagcggag cgtttttgtc aagaagcttt aaagttagaa agattacctt
     3481 ctagtctatt atcatcgcca gaattaaata aattggcaga agaaataaat tatgtaaata
     3541 cagggagtat gaaagactct tctatgcaac caataagctc gcttatagag aatcaaaatg
     3601 gtgacaacct atcggatatt cttaattatc atattattaa atggtgtaaa ttatatcttg
     3661 atgacgccgg ggcaagttgg gcgatgccaa atcgagagaa aggtttttat cgtgcgtggc
     3721 aacaccttat tacatttgat ccagcgctca gtaaaactga acgtaaagtt ttaaaagatt
     3781 ggccagaaga tgcccttata gctttaacaa aagcattatc tgaattagga atttctgaat
     3841 ctaacatgca agcttacctt gaagggcact tactttcttt gcctggatgg gcaggaatga
     3901 tacgatggcg ctcgcaacaa tcaattgagg aacaggaact tctcatagaa tatttagcag
     3961 ttcgactttc tatggagcta gccattgtaa aaccgtattt acctttaaaa aatcaaaagg
     4021 ttgagaaaaa agtttcgatc gtcccactta tagcttcttg gatctattgg ggggatattt
     4081 caatagagaa atggttacaa atgtctgcaa ctgaacaaag tgaattattg gcattcgctt
     4141 atcgctttga tgaaaatact cgcaagaaac tttggctaga agcttgggaa cagacgcatg
     4201 cagagcaatt aagggagaag attgcttcta agcaacgtgc aactcatgat aaaaagcgtg
     4261 tagtagctca attagcattt tgtattgatg tacgttcaga accgtttcgt cgccaccttg
     4321 aaaaattagg tccgttcgaa acgtttggaa tagctggttt ctttgggtta ccaattgcga
     4381 ctactgaact aggtagtaat gacagccatc cttctttgcc agttatatta aaaccgaaac
     4441 atcaaataaa agagctaaca gatgaaaatg aatgtaaatc ttatgaacaa cgtaagatgg
     4501 taggcagttc ggtacgttat acatttaaaa cgatgaaaca gaacgtactg acaagtatgt
     4561 tgctacctga agtaagtggt cctttacttg gtttacaaat ggtgacaagg agttttgtgc
     4621 caagaagagt aggtggcttc attcgtaacc ttcgtaaaaa tatgttgcaa aagcctgata
     4681 caactttctc gcttaatcat gttcatgata caaactgtga gataccgatc ggttttacga
     4741 aagaagaaaa agtgaactat gtgcgtcaaa ctttaaaaat ggtgggatta acagaaggtt
     4801 tcgctccttt agttgtaatg tgcggccaca gtagtcaaag tacgaacaat ccttatgctg
     4861 cagcgcttga gtgtggtgcg tgtggcggag cagcgggagg atttaatgca agagttttcg
     4921 ctactttatg taatcttcca gaagtgagag aggcgttgtc tgctgaaggt attaaaattc
     4981 ccgatgacac catttttgca gcagctgaac ataaaacaac agtggatgaa ttagagtgga
     5041 tttacgttcc agaactttcg gaaactgcgc aagaagcatt tgattgtatt gaggccatta
     5101 tgccaaatgt gagccaacat gcaaatagag agcgtttaat gcaattacct cattttaaaa
     5161 cgaaaataaa aaatccgagt aaagaggcac accgatttgc agaagattgg agtgaaatac
     5221 gtccggaatg gggattagca cgtaatgcat cttttattat cgggcaacgt gaattgactc
     5281 aggagtgtga cctagaaggt agggctttcc ttcataatta tgattggaaa caggatgaaa
     5341 gtggcgatat attagctaac atcatagccg gaccaggaac agtagctcag tggatcaatt
     5401 tacaatatta cgcttcaact gtagcccctc attattatgg tagtggaaac aaagcaaccc
     5461 aaactgtaac ggcaggtctt ggtgttatgc aaggaaatgc aagtgacctg ttaccaggct
     5521 taccttggca atccgttatg caatcagatc gtgagacgta tcattcacca cttcgtttat
     5581 taattgtaat tcaagcgcct actaaatata tagaacgttt gctaaataat aatttcacat
     5641 ttcgagaaaa agttcaaaat ggatgggtta gacttgccag tgttgatcca gagggacgtt
     5701 ggaaaaattg gtaaagcgga attcctgcag cccgggggat cccatggtac gcgtgctaga
     5761 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     5821 tgtcggtgaa cgctctcctg agtaggacaa atccgccgcc gctagcagtt cgtagacgga
     5881 aacaaacgca gaatccaagc aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5941 actcttcctt tttcaatatt attgaagcat ttatcagggt tatgtccact gggttcgtgc
     6001 cttcatccgg atatccagga actgttaaga cccactttca catttaagtt gtttttctaa
     6061 tccgcatatg atcaattcaa ggccgaataa gaaggctggc tctgcacctt ggtgatcaaa
     6121 taattcgata gcttgtcgta ataatggcgg catactatca gtagtaggtg tttccctttc
     6181 ttctttagcg acttgatgct cttgatcttc caatacgcaa cctaaagtaa aatgccccac
     6241 agcgctgagt gcatataatg cattctctag tgaaaaacct tgttggcata aaaaggctaa
     6301 ttgattttcg agagtttcat actgtttttc tgtaggccgt gtacctaaat gtacttttgc
     6361 tccatcgcga tgacttagta aagcacatct aaaactttta gcgttattac gtaaaaaatc
     6421 ttgccagctt tccccttcta aagggcaaaa gtgagtatgg tgcctatcta acatctcaat
     6481 ggctaaggcg tcgagcaaag cccgcttatt ttttacatgc caatacaatg taggctgctc
     6541 tacacctagc ttctgggcga gtttacgggt tgttaaacct tcgattccga cctcattaag
     6601 cagctctaat gcgctgttaa tcactttact tttatctaat ctagacatac tcttcctttt
     6661 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6721 tatttagaaa aataaacaaa taggggttcc gcgtttccct cgacaattcg cgctagacct
     6781 agggcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6841 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6901 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6961 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7021 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7081 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7141 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7201 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7261 acttatcgcc actggcagca gccactggt
//
