LOCUS       sequence-263095-        4107 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFA carrying a Synechococcus elongatus PCC 7942 sbtA gene (Uniprot: 
            Q6UP27).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4107)
  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 4107)
  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-sbtA
FEATURES             Location/Qualifiers
     source          1..4107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(453..547)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(568..1227)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1142..1161
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1228..1330)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1396..1414
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1435..1453,1454..1459,1460..1476,1477..1482,
                     1483..1508)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1435..1453
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1460..1478
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1518..1529
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2655..2672
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2734..2820
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2993..3616)
                     /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     3524..3543
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        complement(3617..3721)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3767..3784)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(3901..4107,1..339))
                     /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(3981..4000)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag tccgaccgct
       61 gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca aaagcaccac
      121 tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg cgccggttaa
      181 ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt acctcggttc
      241 aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt
      301 tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaatcag
      361 ataaaatatt tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcagc
      421 cccatacgat ataagttgtt actagtgctt ggattctcac caataaaaaa cgcccggcgg
      481 caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac
      541 aggagtccaa gcgagctcga tatcaaatta cgccccgccc tgccactcat cgcagtactg
      601 ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg
      661 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac
      721 gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca
      781 gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt
      841 ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg
      901 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg
      961 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgga attccggatg
     1021 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt
     1081 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg
     1141 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt
     1201 ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa
     1261 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     1321 gtgccgatca acgtctcatt ttcgccagat atcgacgtct aagaaaccat tattatcatg
     1381 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcacct cgagtcccta
     1441 tcagtgatag agattgacat ccctatcagt gatagagata ctgagcacat cagcaggacg
     1501 cactgaccga attcattaaa gaggagaaag gtaccatgga tttcttgtcc aatttcttaa
     1561 tggacttcgt taagcagctg cagtctccaa cactcagctt tctgattggc ggcatggtta
     1621 ttgctgcctg tggtagtcag ctgcaaatcc cagagtcgat ttgtaagatc atcgttttca
     1681 tgctgctgac caagattggt ctgaccggcg gtatggcgat ccgcaactcg aatttgacgg
     1741 agatggtcct acccgcgctc ttttcagtag cgattggcat tctgatcgtc ttcatcgccc
     1801 gctacacctt ggcgagaatg ccaaaggtta aaacagtgga tgcgatcgca acgggagggc
     1861 tctttggtgc tgtgagtggc tcgacgatgg ccgcagctct aaccctcctt gaagagcaaa
     1921 aaattcccta cgaagcttgg gccggtgcgc tttatccctt catggatatt ccggccttgg
     1981 tgactgcgat cgtcgtcgcc aacatctatc tcaataagaa gaagcggaaa gaagcagctt
     2041 tcgcgtcagc ccagggggct tacagcaagc agcccgttgc cgctggcgac tattccagct
     2101 cgtcggacta tcccagcagc cggcgcgaat atgctcagca agagtctggg gatcatcggg
     2161 tcaaaatctg gccgattgta gaagaaagcc tgcaaggtcc agccctatca gcaatgttgc
     2221 ttggcgttgc tctgggtctg tttgcccgtc cagaaagtgt ctacgaaggt ttctacgatc
     2281 ctctcttccg ggggctgctt tccatcctga tgctggtcat gggcatggaa gcatggtcga
     2341 gaattagtga gctgcgtaaa gtagctcagt ggtacgtcgt ctacagtatc gtggcgccgc
     2401 tagcccatgg gttcattgcc ttcggccttg gcatgattgc gcactacgca actggtttta
     2461 gcatgggcgg cgttgtggtt ctggcagtca tcgctgcctc tagttccgac atttccggtc
     2521 ccccgacgct gcgcgctggc attccttctg ccaacccctc tgcctacatc ggggcttcca
     2581 ccgcgatcgg aactcctgtg gcaatcggga ttgccattcc gctgttcctt ggccttgccc
     2641 aaaccattgg cggccaccat caccatcacc attaaaatgt aactgtattc agcgatgacg
     2701 aaattcttag ctattgtaat actctagagg catcaaataa aacgaaaggc tcagtcgaaa
     2761 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     2821 ccgccgccgc tagcagttcg tagacggaaa caaacgcaga atccaagcaa aagggaataa
     2881 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     2941 atcagggtta tgtccactgg gttcgtgcct tcatccggat atccaggaac tgttaagacc
     3001 cactttcaca tttaagttgt ttttctaatc cgcatatgat caattcaagg ccgaataaga
     3061 aggctggctc tgcaccttgg tgatcaaata attcgatagc ttgtcgtaat aatggcggca
     3121 tactatcagt agtaggtgtt tccctttctt ctttagcgac ttgatgctct tgatcttcca
     3181 atacgcaacc taaagtaaaa tgccccacag cgctgagtgc atataatgca ttctctagtg
     3241 aaaaaccttg ttggcataaa aaggctaatt gattttcgag agtttcatac tgtttttctg
     3301 taggccgtgt acctaaatgt acttttgctc catcgcgatg acttagtaaa gcacatctaa
     3361 aacttttagc gttattacgt aaaaaatctt gccagctttc cccttctaaa gggcaaaagt
     3421 gagtatggtg cctatctaac atctcaatgg ctaaggcgtc gagcaaagcc cgcttatttt
     3481 ttacatgcca atacaatgta ggctgctcta cacctagctt ctgggcgagt ttacgggttg
     3541 ttaaaccttc gattccgacc tcattaagca gctctaatgc gctgttaatc actttacttt
     3601 tatctaatct agacatactc ttcctttttc aatattattg aagcatttat cagggttatt
     3661 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     3721 gtttccctcg acaattcgcg ctagacctag ggatatattc cgcttcctcg ctcactgact
     3781 cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt
     3841 tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt
     3901 tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg
     3961 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg
     4021 ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc
     4081 tcattccacg cctgacactc agttccg
//
