LOCUS       sequence-324207-        7152 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DabAB2 inorganic carbon transport complex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7152)
  AUTHORS   Flamholz AI, Dugan E, Blikstad C, Gleizer S, Ben-Nissan R, Amram S, 
            Antonovsky N, Ravishankar S, Noor E, Bar-Even A, Milo R, Savage D
  TITLE     Functional reconstitution of a bacterial CO2 concentrating mechanism
            in E. coli.
  JOURNAL   Elife. 2020 Oct 21;9. pii: 59882. doi: 10.7554/eLife.59882.
  PUBMED    33084575
REFERENCE   2  (bases 1 to 7152)
  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: "Elife."; 
            date: "2020-10-21"; volume: "9. pii: 59882. doi"; pages: " 
            10.7554/eLife.59882"
COMMENT     Sequence Label: pFA-DAB2
FEATURES             Location/Qualifiers
     source          1..7152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(31..576)
                     /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(111..130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(690..784)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(805..1464)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1379..1398
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1465..1567)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1633..1651
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1672..1690,1691..1696,1697..1713,1714..1719,
                     1720..1745)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1672..1690
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1697..1715
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1755..1766
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      6016..6102
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(6275..6898)
                     /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     6806..6825
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        complement(6899..7003)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7049..7066)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca
       61 tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca
      121 ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg
      181 tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt
      241 aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg
      301 ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg
      361 cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc
      421 taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa
      481 gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca
      541 gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata
      601 aaatatttct agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc
      661 atacgatata agttgttact agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa
      721 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg
      781 agtccaagcg agctcgatat caaattacgc cccgccctgc cactcatcgc agtactgttg
      841 taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat
      901 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg
      961 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
     1021 attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
     1081 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
     1141 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
     1201 aacactatcc catatcacca gctcaccgtc tttcattgcc atacggaatt ccggatgagc
     1261 attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt
     1321 tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc
     1381 aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt
     1441 atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc
     1501 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     1561 ccgatcaacg tctcattttc gccagatatc gacgtctaag aaaccattat tatcatgaca
     1621 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcacctcga gtccctatca
     1681 gtgatagaga ttgacatccc tatcagtgat agagatactg agcacatcag caggacgcac
     1741 tgaccgaatt cattaaagag gagaaaggta ccatgactat agagttcagc cacacaaccg
     1801 gtcttctcct gctggccatg cccgcactgc tccttatggc cgctgtaatc aaacccaagc
     1861 aaggtgctgc ttatgcgcgc gcttatcggc aacgggttca atggaccagt tttgcggcct
     1921 ttggcctggc cctgctcgct gtcgtcagtt ttctatttgc aaggcaacag aatctgatgc
     1981 tcggtgccgg tagcctccct gccggattgg gattattggc gctttcaatt caagtcaatg
     2041 gcctgactct ggtgctggca agtctggtta gttttgtgct tagcgtgatt gcccgctaca
     2101 gcgtgcagta tctcgatggt gatccccagc aggcccgctt tttccgcttg ctggcggtca
     2161 ccggcggatt tttcctgttg gtcgtcatat caggcaattt gggattattc accctcgcca
     2221 tcattgcgac cggttttggt ttgcatcgac tgctttcttt ttatgccgat cggccgcgcg
     2281 cgatcatggc cacacataaa aaatcaattt tcagccggac tgccgacgcc cttttgctcg
     2341 cggccaccgt cttgatcggt catcaaatcg gtagtttgga gttcagccaa atcagcgcct
     2401 atgttcatgc gcaagaccac ctatccattg cgttgcacgt ggctgcttgg ctgatcgtgc
     2461 tggccgccat ccttaaaagc gcgcaatttc cgtttcatgg ctggctgatt caggtcatgg
     2521 aagcacccac gccggtttcc gcgctcatgc acgccggtgt cgtttacagc ggggcaatca
     2581 ttgtgctgcg caccagcgaa ttattggccg ccgacggtac cgccctgttg cttctggccc
     2641 tgattggttt gatgaccttg gctatcggct cgttggtcat gctgacccaa tcggcgatca
     2701 aatcatcact ggcctggtct accgccgctc agctcggctt catgatgctc gaactgggtt
     2761 tggggttgtt tggcttggcc ttgctgcact tggttggcca ctcgctctac aaggcccatg
     2821 cgttcttgtc ctcgggcagc atgaccgatc acctgcgtca ggccaaagtg ctcaaaaacc
     2881 gcccaatctc tgttgtggcc tggtttacca cggtgattgt gagcggcttg ttcaccctcg
     2941 gtatcgcggc ggcgatgggt ttatcgatcg atcaggagcc gatgttgccc gccgtgctga
     3001 ccatcatcgc cttggcgacc gctcagttga tgctcaaggc attgagccat ggcacatggc
     3061 gtgaaattct ggtcgccgca ggcgcggcaa tcgccatgac cggcgtgtat gtcttcctgc
     3121 atgaggtgtt catcacaggc tttgccgaca cgctggccgc gacaccgcaa cgtgcgccgc
     3181 tgctcgatct gctgttgatg gcgatcacga tcataacctt cctgtttgtt gcctggcttc
     3241 aaggccccgg caaaaccttg atgagcccgg agcgacaatt cgccttgttt gttcacctca
     3301 ataacgggct gtatctcgac cgctgggtcg aacgcttggc cttccgtttc tggccggaga
     3361 aagttggccg cgcaccgaaa aaatcctgcg ccgtgattcc gcctaaccca tccggtatcg
     3421 agccgtaaat ccctgccaca aaggcacaca ccatttaccc gaggatttcg accatgacga
     3481 ccctgacatc attacaacgc agcgaagcgc aacgaaacca cattgttgac ctgattgaca
     3541 aagcctgcct gcgcatcgcg cccatctggc cactggattc gtttgtcgcc gtaaacccct
     3601 atcttggttt gattgatcaa ccctttgata cggtgggtcg gtatctggaa caaaccgtgg
     3661 gcgaatccct atttatggat cacggctggt ttgccgacaa gatcgcccaa ggggaaatca
     3721 ccgatgacga tttggcgcag gcggcgcagc aactggaccc atcaattagc cttgatacca
     3781 tcaagcagca actggccgtc catcgacaac ctgcacccgc gctgcctttg gtcacaaacg
     3841 agctcgatcg ccgcgacgcg ccgccagtat ctgaatttgt catcgagcaa gtcagccagt
     3901 tcatggccaa ctactatgat cgcgggcaag ccctctggca cttacccaaa gaagccagtg
     3961 ccagtttatt cgcccaatgg cgccgctaca ccttgattga ccgcagcgcc agcgccgtcg
     4021 gcttaaagca ggttcgccaa cacttgcttg ccgtcccaag cgatgccatt gatgcattgt
     4081 tctgggcgct ggaccagatc aatctgcccg aaagtcgcct gcccgactac ctgttcacct
     4141 tattaaaaac catcggtggc tgggcgagct ggtgccgcta tctacacttt caagccggtt
     4201 tgcacggcga gtcgcaacac gatttacgcg atttgctgat cattcgcttg gtctgggttg
     4261 ctttggtgat caaagaaacc agctcggctg gacggcaaca atggcgcgcg aagctaaacg
     4321 attggtttga tccggccaaa ttggtcgcaa gcccttcggc gacagctact gcttcgacta
     4381 aggctcaatc cagccggatc gatgaaatcc tgctggcggc cgccgagcag gcgtttcgac
     4441 gccggatcaa tgccggcttg aaccgccagc ctgccgatgc acccgatcag caagccgaac
     4501 gcccgactgt tcaggccgcg ttttgcattg acgtgcgttc cgaggtgttc cgccgccacc
     4561 tcgaagccag cagcccgggg ctcgaaacca tcggttttgc cggttttttc ggcctgccga
     4621 ttgattattg ccgcatgggc gaatcagagg ctcggctaca aaacccggtg ctcatcaacc
     4681 ccgcctaccg tgcgcaggag acgggcgatc cggccattgc ccaacatcgc catgcccgac
     4741 aaagccgagg ggccatctgg aagcaattca aactttccgc cgcctcgtgc ttcacctttg
     4801 tcgaatccgc aggcttgagc tacgtgccgc gtttgctggc cgattcactg ggttggcatc
     4861 gctcatcgct gccgcctgat gcccccgggc ttacccctga ggagcgtgcg cgtttgcatc
     4921 cccaattggt caaactcgat ggcggcgcac tatcgacgca ggaaaaagtc gatctggccg
     4981 aaaaagtact ccgcgggctg gggttaacgc acacctttgc gcccatcgtc ctgctcgccg
     5041 gacacggcag ctccaccacc aacaacccgc atcgcgcggg gttggattgc ggtgcctgcg
     5101 ccggtcaggc aggcgatgtg aatgctcgcg tcgcggtgca gttgctcaat gaggccgccg
     5161 tgcgactggg gctgatcgaa cggggtattg ccatcccgcg tgacacccgg ttcgtggccg
     5221 ccctgcacga caccaccaca gaccacatcg aactactgga tttggatcaa tcaggcatcg
     5281 agagtgatca attgtcatcc ttgacccagg cgctcaaaca agccggtgag ctcactcgtc
     5341 tggagcgttt ggtgacgctt gaggcccaag tcgataccgt tgatgccgaa aaacaggcca
     5401 cgttccgagg gcgagactgg tcgcaagtac ggccggaatg ggggctagcc ggcaatgctg
     5461 ccttcatcgc cgcaccccgc tggcgcacgc gcgggttgga tcttggcggc cgcgccttcc
     5521 tgcatgatta cgattggcga cacgataagg agtttggcgt tttgaacgtc atcatgaccg
     5581 cgcccttgat cgtcgcaaac tggatcaacc tgcaatacta cggctccacg gtcgataatc
     5641 tgcatcaagg ggcgggcaat aaagtgctgc ataatgtggt cggcggcacc gtcggggtga
     5701 ttgaaggcaa tggcggcgac ctgcgagttg gtcttgccat gcaatcgctg cacgatggtg
     5761 aacagtggcg gcacgaaccg ctgcggctct cggcgtacat tgaggcgccc attgcagaaa
     5821 tcgataaaat tattgccggt cacgacatgc tgaacgcgtt aatcaacaac cgctggatgc
     5881 acatcctgca tatcgatgac aacggcatcc cgcatcgccg tcatgcccac ggtgactggc
     5941 gaccggaacc gatctgaaat gtaactgtat tcagcgatga cgaaattctt agctattgta
     6001 atactctaga ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     6061 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccgcc gctagcagtt
     6121 cgtagacgga aacaaacgca gaatccaagc aaaagggaat aagggcgaca cggaaatgtt
     6181 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tatgtccact
     6241 gggttcgtgc cttcatccgg atatccagga actgttaaga cccactttca catttaagtt
     6301 gtttttctaa tccgcatatg atcaattcaa ggccgaataa gaaggctggc tctgcacctt
     6361 ggtgatcaaa taattcgata gcttgtcgta ataatggcgg catactatca gtagtaggtg
     6421 tttccctttc ttctttagcg acttgatgct cttgatcttc caatacgcaa cctaaagtaa
     6481 aatgccccac agcgctgagt gcatataatg cattctctag tgaaaaacct tgttggcata
     6541 aaaaggctaa ttgattttcg agagtttcat actgtttttc tgtaggccgt gtacctaaat
     6601 gtacttttgc tccatcgcga tgacttagta aagcacatct aaaactttta gcgttattac
     6661 gtaaaaaatc ttgccagctt tccccttcta aagggcaaaa gtgagtatgg tgcctatcta
     6721 acatctcaat ggctaaggcg tcgagcaaag cccgcttatt ttttacatgc caatacaatg
     6781 taggctgctc tacacctagc ttctgggcga gtttacgggt tgttaaacct tcgattccga
     6841 cctcattaag cagctctaat gcgctgttaa tcactttact tttatctaat ctagacatac
     6901 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6961 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgtttccct cgacaattcg
     7021 cgctagacct agggatatat tccgcttcct cgctcactga ctcgctacgc tcggtcgttc
     7081 gactgcggcg agcggaaatg gcttacgaac ggggcggaga tttcctggaa gatgccagga
     7141 agatacttaa ca
//
