LOCUS       sequence-263329-        3648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFA carrying a E. coli canA gene (Uniprot: P61517).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3648)
  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 3648)
  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: pFA-eCA
FEATURES             Location/Qualifiers
     source          1..3648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..548)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(83..102)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(662..756)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(777..1436)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1351..1370
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1437..1539)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1605..1623
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1644..1662,1663..1668,1669..1685,1686..1691,
                     1692..1717)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1644..1662
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1669..1687
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1727..1738
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1748..1765
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2484..2570
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2743..3366)
                     /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     3274..3293
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        complement(3367..3471)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3517..3534)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg
       61 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg
      121 cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg
      181 tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat
      241 gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      301 caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg
      361 agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact
      421 gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa
      481 aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac
      541 gatctcaaga agatcatctt attaatcaga taaaatattt ctagatttca gtgcaattta
      601 tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtta ctagtgcttg
      661 gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga
      721 gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcgat atcaaattac
      781 gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg
      841 aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct
      901 tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg
      961 tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca
     1021 ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat
     1081 atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca
     1141 gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg
     1201 tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag aatgtgaata
     1261 aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc
     1321 agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct
     1381 ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta
     1441 gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt
     1501 tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattt tcgccagata
     1561 tcgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     1621 ggccctttcg tcttcacctc gagtccctat cagtgataga gattgacatc cctatcagtg
     1681 atagagatac tgagcacatc agcaggacgc actgaccgaa ttcattaaag aggagaaagg
     1741 taccatgcat catcaccatc accacaaaga catagataca ctcatcagca acaatgcact
     1801 atggtcaaaa atgctggtgg aagaggatcc cgggtttttt gagaaactgg cacaagcgca
     1861 aaaaccgcgc tttctatgga ttggatgttc cgacagtcgc gttcctgcag aacgtttaac
     1921 cggtcttgag ccgggcgaac tctttgttca ccgtaatgtt gctaacctgg tcattcacac
     1981 tgacctgaac tgcctttccg tggttcagta tgcagtggat gtactcgaag ttgaacacat
     2041 tattatctgt ggccactacg gttgcggcgg cgtacaagcc gcagttgaaa acccggaact
     2101 ggggcttatc aacaactggc tgctgcatat ccgcgatatc tggttcaaac atagctcatt
     2161 gctcggcgaa atgccgcaag agcgccgtct ggataccttg tgtgaactga acgtcatgga
     2221 acaggtgtat aacctgggcc actccaccat tatgcaatca gcgtggaaac gcgggcagaa
     2281 agttaccatt cacggctggg cctacggcat tcacgacggc ttgctgcgtg atctggatgt
     2341 taccgccacc aaccgcgaaa cccttgagca acgttaccgt cacgggattt ccaacctcaa
     2401 gctgaaacac gccaaccaca aataaaatgt aactgtattc agcgatgacg aaattcttag
     2461 ctattgtaat actctagagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     2521 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgccgc
     2581 tagcagttcg tagacggaaa caaacgcaga atccaagcaa aagggaataa gggcgacacg
     2641 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2701 tgtccactgg gttcgtgcct tcatccggat atccaggaac tgttaagacc cactttcaca
     2761 tttaagttgt ttttctaatc cgcatatgat caattcaagg ccgaataaga aggctggctc
     2821 tgcaccttgg tgatcaaata attcgatagc ttgtcgtaat aatggcggca tactatcagt
     2881 agtaggtgtt tccctttctt ctttagcgac ttgatgctct tgatcttcca atacgcaacc
     2941 taaagtaaaa tgccccacag cgctgagtgc atataatgca ttctctagtg aaaaaccttg
     3001 ttggcataaa aaggctaatt gattttcgag agtttcatac tgtttttctg taggccgtgt
     3061 acctaaatgt acttttgctc catcgcgatg acttagtaaa gcacatctaa aacttttagc
     3121 gttattacgt aaaaaatctt gccagctttc cccttctaaa gggcaaaagt gagtatggtg
     3181 cctatctaac atctcaatgg ctaaggcgtc gagcaaagcc cgcttatttt ttacatgcca
     3241 atacaatgta ggctgctcta cacctagctt ctgggcgagt ttacgggttg ttaaaccttc
     3301 gattccgacc tcattaagca gctctaatgc gctgttaatc actttacttt tatctaatct
     3361 agacatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3421 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gtttccctcg
     3481 acaattcgcg ctagacctag ggatatattc cgcttcctcg ctcactgact cgctacgctc
     3541 ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga
     3601 tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccg
//
