LOCUS       sequence-263091-        7196 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pFE carrying the vcdabA2 ( locus_tag: CSW01_RS07960) and vcdabB2 
            genes (locus_tag: CSW01_RS07955).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7196)
  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 7196)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-vcDAB
FEATURES             Location/Qualifiers
     source          1..7196
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(16..110)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(141..935)
                     /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(252..271)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     862..881
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1114..1132
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1153..1171,1172..1177,1178..1194,1195..1200,
                     1201..1226)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1153..1171
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1178..1196
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1236..1247
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      5373..5459
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5632..6255)
                     /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     6163..6182
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        complement(6256..6360)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6536..7124)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6616..6635)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgactagtg cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa
       61 caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct
      121 ctcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc
      181 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca
      241 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc
      301 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag
      361 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg
      421 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca
      481 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat
      541 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact
      601 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc
      661 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc
      721 gtggccagcc acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg
      781 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca
      841 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc
      901 ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct
      961 tgatcagatc ttgatcccct gcgccatcag atccttggcg gcaagaaagc catccagttt
     1021 actttgcagg gcttcccaac cttaccagag ggcgccccag ctggcaattc cgacgtctaa
     1081 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     1141 cttcacctcg agtccctatc agtgatagag attgacatcc ctatcagtga tagagatact
     1201 gagcacatca gcaggacgca ctgaccgaat tcattaaaga ggagaaaggt accgtccatg
     1261 tcggaactgt caccccagtg gctccttgtt agtttgtatc ttggtggcct tgtattgagc
     1321 ttatggttac ctcaacagca gtggcgtatc gccaagttcg ccactctcgc cggcttcatc
     1381 gccatgcttg gaaataccct ttttgcgata gggcaagcga gcatcaacct agtcccagtc
     1441 gatggattag gcatcgtgat ggcgatactc gtctcgctcc taggctgggt gataacccgc
     1501 ttcgcccatc gttatctgca aggcgaagcg cagcaagcgc gttttgtcac cgccacgctg
     1561 tttaccctaa gctgcgtagc cttgttggtc accagtcagc atctgcttgt gatcgtgctg
     1621 gcgtggagcg gaaccagcgt tggcctacat cgtctcctga ccttttaccc agagcgcaaa
     1681 gccgcacgcg tggtggcaca caaaaaattc ttagtcagcc gttttgccga aatctgtttg
     1741 ttgttagcgc tcggtttgat ctatctcgat gtcggtagct tgtcgctaca agagattcac
     1801 ttcgcgttaa acgcgcagga ttcgctgcca cttagcttaa caggggctgc gttgttactc
     1861 gccttcgctg ctatcttaaa aaccgcccag ttacctctgc acggctggct gattcaggtg
     1921 atggaagcgc caacgccagt ctcggcgtta ttgcatgcgg gtgtggttaa catgggggga
     1981 ttcgttctgc tgcgcgtcgc agaactgatt ggacttgttc cttcggcgca atggttgctg
     2041 gtgatcgtcg gtagcctcac tgcggtgctc gctggcatgg tcatgctgac tcggatcagc
     2101 attaaagttc gtttagcttg gtcaacgtgc gcgcaaatgg gctttatgct gatggaaatc
     2161 ggcttaggct tatatgagct cgccctgctc catttggtgg cgcattcact ttataaagct
     2221 tatgccttcc taagttctgg cgaagcggtg gagcgagcac gtctgcatga ttttcaagcc
     2281 ccacaaagtg tcaatggtaa agccgtgggg gctttgctcg cggtagggct cagcgcagcg
     2341 cttatcctga tactgcacca tttatggcaa tccttaacat ccaatttgca tctacccaat
     2401 gcggtggtcg cgattctcgc cttaggtttt gctccgctac tttggcaaag tggacgtttt
     2461 acaccaagtg gcttgagcct cggtgtgctg cgggttgccc tactcaccca actctatttc
     2521 gtatggcatc tgctgtttgc gggtttagca cccgagctgc gccccgctag cccactcgcg
     2581 ctgagctttg tgatgttctg ttttatcacc ttgtatctga tgcaagtgct gctctgcctc
     2641 taccccaaaa gccgtttatc actgtggctc tatccttggg tttaccatgg tttttatctt
     2701 gatgagactt ttacgcgcct gaccttcaaa tattggcccg tgcaattaac tctgcctacc
     2761 gctctttcat caactcaagc gcaaccgacg acaggagaac gcgtatgaat gcgccacgtt
     2821 cgctctcagc tttaagcacc accgctaacc cagcattatc gacacttgtc gaacaagtct
     2881 gcgccttgat tgctcctaac tggccgctgg atcgcatgat tgcggtgagc ccttattgga
     2941 aacggattga taagcccttt gcacaagccg ccgccgagct caaacaattg gccgcctccc
     3001 ccatgaccat gacgctcagt gattaccacc ttcgttggca aaataagcaa atccaatcag
     3061 cggatttgca acaagccatc gctgaacaaa acagtgacct tagcgaatct accttgattg
     3121 ccgctttaca acagcccaca gcgccgagtc atccttggcc attgctgtgc gacaccgtcg
     3181 atagccggcg tgatcttgag catcatcccg catggaatga cgccatcacc catcaaattt
     3241 cgcaattttg cgccgcctat tttgaccatc atcaagccga ttggagccca gatcagcaaa
     3301 ccggactctt tgccacttgg cgtgaggcga tgatccacga tcgcagcatt acgctactgc
     3361 tcaacgagac aagcgtaaaa caaaaagcca ccaagcttcc cgaggatgct atggccgcca
     3421 tcgagcaaac tctagcgcaa ctcgctattg cccctgcgca acaagaaacg tatttacaag
     3481 ccgtgttaat gcgcatcagt ggctgggcat cctggtgtgc ttatctcgca tggcaggcag
     3541 ggtttgaggg tcggcacgat gagcacctgc gcgatctgtt ggcgattcgt ctctgctggg
     3601 agaacttact tgatgatggt gagcgcggta tgggctctgt gtggttacaa tggcagcaat
     3661 cttgggcacc gcgccaatcg tgcgaggagg atcgcgcact acgcattgcg ctactgtggc
     3721 aacgcagtgc ggaaatcgcc tatcaacgcc aactttttgc tgcgctgaca tcagcacaag
     3781 agtcggctcc ccaatcaagt tatcccgagg tgcaagcggc tttttgtatc gatgtgcgtt
     3841 cagaagtcat tcgccgccac ttagaagcgc aatcacccca tattcaaacc cttggcttcg
     3901 caggcttttt tggcttaccg atccgctatc agttattggg caccgaagca agccgcccac
     3961 agttaccggg cttattagcg ccgagcttga tcgtctctga cagcacaggg aatgaagatc
     4021 aagacgcgaa attggccttg cgtcgccgtg ctcgactcaa gcgacacttt agctggcgcg
     4081 cgtttcatca cctccccgcc tccacgttta ctctggtgga aaccacgggg ttggcgtatt
     4141 taaccaagct ccttaaacgc accttatcct accctgcatt gagtgcatcg gtggagcgtt
     4201 tcgcctttac ggaacatgag tggcagagtg tcaaaccgca attcacgcgc gatccacaga
     4261 cgttagcgca gcgtgctcaa atggcggcca acattctgcg agcgttgggg atcgccaccg
     4321 agcaagcacg cttggtgcta ctcgtggggc acggctcaca gacgcaaaat aatccgcagc
     4381 gtgcagggct ggattgtgga gcctgctgcg gtcaaagcgg cgaagtcaac gctcgcacgt
     4441 tagcagcatt gctaaatgat caggcggtgc gccaagcgtt gcctgaatat ggaatttcgc
     4501 tgcgcgatga cgtccacttt atcgcggcgc tacacaacac cacgacagaa gcgatgaggc
     4561 tttttgatcg ccatgagatc ccaacaagcc atcgcgaggc gctagaacaa cttgatcagc
     4621 agctcacggc cgccagccat ggcgcacgtc aagaacgcgc gccaagtctt gaattgaatc
     4681 acaaccacca agcgcctcca agcaaggaca acgccctatc tgcccagcag ttggagcaag
     4741 cgtttctgcg tcgagcccat gattgggcgc aaactcgccc tgaatggggg ctgaccaaca
     4801 atgccgcttt tatcattgca ccgcgccaac gttcaaagca ggcgaaactc gatggtcgag
     4861 tgtttttgca tgaataccaa ccggagcgcg atcccgaagg gcaattactg acacaaatta
     4921 tgacagcccc aatgctggtc acacactgga ttaacatgca gtattttgcc tccaccgtcg
     4981 ataatcgccg cttcggcagt ggcaataaaa ccttgcataa tgtggtagga ggaaatatcg
     5041 gtctttttga aggcaacgga ggcgatctgc gctgtggctt agccctgcaa tcgctgcatg
     5101 atggccaagg gtggcgacat gaagccttac gtctcactgt ggtgattgat gcgccgcgcg
     5161 aacggattga gcaagtgatg gcttcacacc gcgtggttga acatttggtt aagcatgaat
     5221 ggctctatct ggcccgtttt gctgatcaag ggattgagat atatctccaa ggcacttggc
     5281 aacgcatcac tcagcccagc tcagattcca gtgcgagata aagcggaatt cctgcagccc
     5341 gggggatccc atggtacgcg tgctagaggc atcaaataaa acgaaaggct cagtcgaaag
     5401 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
     5461 cgccgccgct agcagttcgt agacggaaac aaacgcagaa tccaagcaaa agggaataag
     5521 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5581 tcagggttat gtccactggg ttcgtgcctt catccggata tccaggaact gttaagaccc
     5641 actttcacat ttaagttgtt tttctaatcc gcatatgatc aattcaaggc cgaataagaa
     5701 ggctggctct gcaccttggt gatcaaataa ttcgatagct tgtcgtaata atggcggcat
     5761 actatcagta gtaggtgttt ccctttcttc tttagcgact tgatgctctt gatcttccaa
     5821 tacgcaacct aaagtaaaat gccccacagc gctgagtgca tataatgcat tctctagtga
     5881 aaaaccttgt tggcataaaa aggctaattg attttcgaga gtttcatact gtttttctgt
     5941 aggccgtgta cctaaatgta cttttgctcc atcgcgatga cttagtaaag cacatctaaa
     6001 acttttagcg ttattacgta aaaaatcttg ccagctttcc ccttctaaag ggcaaaagtg
     6061 agtatggtgc ctatctaaca tctcaatggc taaggcgtcg agcaaagccc gcttattttt
     6121 tacatgccaa tacaatgtag gctgctctac acctagcttc tgggcgagtt tacgggttgt
     6181 taaaccttcg attccgacct cattaagcag ctctaatgcg ctgttaatca ctttactttt
     6241 atctaatcta gacatactct tcctttttca atattattga agcatttatc agggttattg
     6301 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6361 tttccctcga caattcgcgc tagacctagg gcgttcggct gcggcgagcg gtatcagctc
     6421 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6481 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6541 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6601 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6661 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6721 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6781 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6841 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6901 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6961 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7021 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     7081 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     7141 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtc
//
