LOCUS       sequence-263330-        5616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFE carrying a dabA2 gene (Uniprot: D0KWS7).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5616)
  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 5616)
  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-dabA2
FEATURES             Location/Qualifiers
     source          1..5616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      52..138
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(311..934)
                     /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     842..861
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        complement(935..1039)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1215..1803)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1295..1314)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1891..1985)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2016..2810)
                     /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(2127..2146)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2737..2756
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2989..3007
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(3028..3046,3047..3052,3053..3069,3070..3075,
                     3076..3101)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    3028..3046
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3053..3071
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             3111..3122
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 agcggaattc ctgcagcccg ggggatccca tggtacgcgt gctagaggca tcaaataaaa
       61 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
      121 ctcctgagta ggacaaatcc gccgccgcta gcagttcgta gacggaaaca aacgcagaat
      181 ccaagcaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
      241 aatattattg aagcatttat cagggttatg tccactgggt tcgtgccttc atccggatat
      301 ccaggaactg ttaagaccca ctttcacatt taagttgttt ttctaatccg catatgatca
      361 attcaaggcc gaataagaag gctggctctg caccttggtg atcaaataat tcgatagctt
      421 gtcgtaataa tggcggcata ctatcagtag taggtgtttc cctttcttct ttagcgactt
      481 gatgctcttg atcttccaat acgcaaccta aagtaaaatg ccccacagcg ctgagtgcat
      541 ataatgcatt ctctagtgaa aaaccttgtt ggcataaaaa ggctaattga ttttcgagag
      601 tttcatactg tttttctgta ggccgtgtac ctaaatgtac ttttgctcca tcgcgatgac
      661 ttagtaaagc acatctaaaa cttttagcgt tattacgtaa aaaatcttgc cagctttccc
      721 cttctaaagg gcaaaagtga gtatggtgcc tatctaacat ctcaatggct aaggcgtcga
      781 gcaaagcccg cttatttttt acatgccaat acaatgtagg ctgctctaca cctagcttct
      841 gggcgagttt acgggttgtt aaaccttcga ttccgacctc attaagcagc tctaatgcgc
      901 tgttaatcac tttactttta tctaatctag acatactctt cctttttcaa tattattgaa
      961 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     1021 aacaaatagg ggttccgcgt ttccctcgac aattcgcgct agacctaggg cgttcggctg
     1081 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1141 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1201 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1261 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1321 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1381 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     1441 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1501 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1561 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1621 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     1681 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1741 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1801 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     1861 taagggattt tggtcatgac tagtgcttgg attctcacca ataaaaaacg cccggcggca
     1921 accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag
     1981 gagtccaagc gagctctcga accccagagt cccgctcaga agaactcgtc aagaaggcga
     2041 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
     2101 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
     2161 cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
     2221 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
     2281 cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
     2341 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
     2401 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     2461 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
     2521 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
     2581 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg cagttcattc
     2641 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
     2701 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     2761 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
     2821 cctcatcctg tctcttgatc agatcttgat cccctgcgcc atcagatcct tggcggcaag
     2881 aaagccatcc agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc
     2941 aattccgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     3001 acgaggccct ttcgtcttca cctcgagtcc ctatcagtga tagagattga catccctatc
     3061 agtgatagag atactgagca catcagcagg acgcactgac cgaattcatt aaagaggaga
     3121 aaggtaccgt ccatgacgac cctgacatca ttacaacgca gcgaagcgca acgaaaccac
     3181 attgttgacc tgattgacaa agcctgcctg cgcatcgcgc ccatctggcc actggattcg
     3241 tttgtcgccg taaaccccta tcttggtttg attgatcaac cctttgatac ggtgggtcgg
     3301 tatctggaac aaaccgtggg cgaatcccta tttatggatc acggctggtt tgccgacaag
     3361 atcgcccaag gggaaatcac cgatgacgat ttggcgcagg cggcgcagca actggaccca
     3421 tcaattagcc ttgataccat caagcagcaa ctggccgtcc atcgacaacc tgcacccgcg
     3481 ctgcctttgg tcacaaacga gctcgatcgc cgcgacgcgc cgccagtatc tgaatttgtc
     3541 atcgagcaag tcagccagtt catggccaac tactatgatc gcgggcaagc cctctggcac
     3601 ttacccaaag aagccagtgc cagtttattc gcccaatggc gccgctacac cttgattgac
     3661 cgcagcgcca gcgccgtcgg cttaaagcag gttcgccaac acttgcttgc cgtcccaagc
     3721 gatgccattg atgcattgtt ctgggcgctg gaccagatca atctgcccga aagtcgcctg
     3781 cccgactacc tgttcacctt attaaaaacc atcggtggct gggcgagctg gtgccgctat
     3841 ctacactttc aagccggttt gcacggcgag tcgcaacacg atttacgcga tttgctgatc
     3901 attcgcttgg tctgggttgc tttggtgatc aaagaaacca gctcggctgg acggcaacaa
     3961 tggcgcgcga agctaaacga ttggtttgat ccggccaaat tggtcgcaag cccttcggcg
     4021 acagctactg cttcgactaa ggctcaatcc agccggatcg atgaaatcct gctggcggcc
     4081 gccgagcagg cgtttcgacg ccggatcaat gccggcttga accgccagcc tgccgatgca
     4141 cccgatcagc aagccgaacg cccgactgtt caggccgcgt tttgcattga cgtgcgttcc
     4201 gaggtgttcc gccgccacct cgaagccagc agcccggggc tcgaaaccat cggttttgcc
     4261 ggttttttcg gcctgccgat tgattattgc cgcatgggcg aatcagaggc tcggctacaa
     4321 aacccggtgc tcatcaaccc cgcctaccgt gcgcaggaga cgggcgatcc ggccattgcc
     4381 caacatcgcc atgcccgaca aagccgaggg gccatctgga agcaattcaa actttccgcc
     4441 gcctcgtgct tcacctttgt cgaatccgca ggcttgagct acgtgccgcg tttgctggcc
     4501 gattcactgg gttggcatcg ctcatcgctg ccgcctgatg cccccgggct tacccctgag
     4561 gagcgtgcgc gtttgcatcc ccaattggtc aaactcgatg gcggcgcact atcgacgcag
     4621 gaaaaagtcg atctggccga aaaagtactc cgcgggctgg ggttaacgca cacctttgcg
     4681 cccatcgtcc tgctcgccgg acacggcagc tccaccacca acaacccgca tcgcgcgggg
     4741 ttggattgcg gtgcctgcgc cggtcaggca ggcgatgtga atgctcgcgt cgcggtgcag
     4801 ttgctcaatg aggccgccgt gcgactgggg ctgatcgaac ggggtattgc catcccgcgt
     4861 gacacccggt tcgtggccgc cctgcacgac accaccacag accacatcga actactggat
     4921 ttggatcaat caggcatcga gagtgatcaa ttgtcatcct tgacccaggc gctcaaacaa
     4981 gccggtgagc tcactcgtct ggagcgtttg gtgacgcttg aggcccaagt cgataccgtt
     5041 gatgccgaaa aacaggccac gttccgaggg cgagactggt cgcaagtacg gccggaatgg
     5101 gggctagccg gcaatgctgc cttcatcgcc gcaccccgct ggcgcacgcg cgggttggat
     5161 cttggcggcc gcgccttcct gcatgattac gattggcgac acgataagga gtttggcgtt
     5221 ttgaacgtca tcatgaccgc gcccttgatc gtcgcaaact ggatcaacct gcaatactac
     5281 ggctccacgg tcgataatct gcatcaaggg gcgggcaata aagtgctgca taatgtggtc
     5341 ggcggcaccg tcggggtgat tgaaggcaat ggcggcgacc tgcgagttgg tcttgccatg
     5401 caatcgctgc acgatggtga acagtggcgg cacgaaccgc tgcggctctc ggcgtacatt
     5461 gaggcgccca ttgcagaaat cgataaaatt attgccggtc acgacatgct gaacgcgtta
     5521 atcaacaacc gctggatgca catcctgcat atcgatgaca acggcatccc gcatcgccgt
     5581 catgcccacg gtgactggcg accggaaccg atctga
//
