LOCUS       sequence-202794-        5607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCKTRBS-OD derivative carrying CbnR-ABE44898-OD chimera, also known 
            as ChBz01.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5607)
  AUTHORS   Juarez JF, Lecube-Azpeitia B, Brown SL, Johnston CD, Church GM
  TITLE     Biosensor libraries harness large classes of binding domains for 
            construction of allosteric transcriptional regulators.
  JOURNAL   Nat Commun. 2018 Aug 6;9(1):3101. doi: 10.1038/s41467-018-05525-6.
  PUBMED    30082754
REFERENCE   2  (bases 1 to 5607)
  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 
            Commun."; date: "2018-08-6"; volume: "9(1):3101. doi"; pages: " 
            10.1038/s41467-018-05525-6"
COMMENT     Sequence Label: pCKTRBS-CbnR-ABE44898-OD
FEATURES             Location/Qualifiers
     source          1..5607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..19,20..25,26..42,43..48,49..74)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1..19
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    26..44
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             113..121
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             complement(2027..2686)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2601..2620
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2687..2789)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      3134..3356
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             3404..4354
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     protein_bind    4880..4901
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(4916..5542)
                     /codon_start=1
                     /gene="tetR"
                     /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="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     5447..5466
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 tccctatcag tgatagagat tgacatccct atcagtgata gagatactga gcacatcagc
       61 aggacgcact gaccggcggc cgcgcgaatt cgagctcggt acccgggtga cctaaggagg
      121 taaataatgg aattccggca gctcaagtat ttcatcgccg tcgcggaagc agggaacatg
      181 gctgcagcag ccaagcgcct gcacgtctca cagcccccca tcacgcggca gatgcaagcc
      241 ctggaagcgg acttgggcgt cgtgcttctg gagcgaagcc accgcgggat cgaactcacc
      301 gccgcaggtc acgctttcct ggaggatgcg cgccgcatcc tggagttggc aggccgttcg
      361 ggtgatcgct cccgcgccgc cgcgcggggc atgacctcac gccgacttgt tttgacccgc
      421 agtgctgccg ttatcggcgc cgcatccact ggcctgctgc tgcccgaaat cgtgcgcgca
      481 cagtcgggca aggtgcgtgt gggcttcatg ttgccctaca ccggcacctt tgcccaactc
      541 ggggtggcca ttgaaaacgg ctttcgcctg gcgatcaatg agcaaggcgg caagctgggc
      601 ggccgcgaga tcgaatactt caaggtcgac gatgagtccg agccgtccaa aggcattgaa
      661 aacgccagca agctggtgca acgcgacaag gtcgatgtac tggtcggcac ggtgcactcc
      721 ggtgtgcaga tgggtatcca gaaggtcgcg cgcgacagcg gcgtactgag cctgattccc
      781 aacgcgggcg tgcatgccgc cacccgcagc ctgtgcgcac ccaatgtgtt ccgtacctcc
      841 ttcagcaact cgcaacctac ccgcgccctg ggtcaggcca tgatcgcaaa aggccataaa
      901 aaagccgtct ggatcacctg gaaatatgca gccggcgacg aggcctttga aggcttcaag
      961 gaaagctaca cagcggccgg cggcaccatt atcaaggaac tgggcctgcc cttccccaac
     1021 gttgaattcc aggcgctgct gaccgaaatc gccgcactca agcccgatgc cgtggcctgc
     1081 ttcttcgccg gcggcggcgc cgcgaagttc atccgcgact acgcagcggc gggcctcaaa
     1141 ggcaagatcc cgctctacgg ttccggcttc ctgaccgaag gcgtgctgga cgcggccggc
     1201 cctgcggcgg acggcatcat caccaccatg cactacagcg actcgctgga cacgccgcgc
     1261 aacaagaaat tccgcctcga ctacgccaaa gccttcaaga cgcaacccga tgtgtacgcg
     1321 gtgcagggct atgacaccgg cctgctgctg atccagggcg ccaacgccgt caagggcgac
     1381 ctggccaaca agcccgcgct ctacaaggcc atggaaggcg ccaccatcga cagcccgcgc
     1441 ggcaagtgga ccatgagcaa ggcacacaac ccggtgcagg acatgtacct gcgtgtggtg
     1501 gaaaacaagg aaaacaaggt gctcggcgtt gcggccaagg cactggccga ctccggcgcc
     1561 ggctgcaaga tgggcaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc
     1621 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     1681 cagtgagatc ctctagagtg gacctgcagg catgcaagct tgcggccgcc cgtttttatc
     1741 aggctctggg aggcagaata aatgatcata tcgtcaatta ttacctccac ggggagagcc
     1801 tgagcaaact ggcctcaggc atttgagaag cacacggtca cactgcttcc ggtagtcaat
     1861 aaaccggtaa accagcaata gacataagcg gctatttaac gaccctgccc tgaaccgacg
     1921 accgggtcga atttgctttc gaatttctgc cattcatccg cttattatca cttattcagg
     1981 cgtagcaacc aggcgtttaa gggcaccaat aactgcctta aaaaaattac gccccgccct
     2041 gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac
     2101 aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
     2161 atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa
     2221 aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt
     2281 tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa
     2341 actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat
     2401 ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg
     2461 ccatacgaaa ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat
     2521 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg
     2581 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc
     2641 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag
     2701 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     2761 gaaagttgga acctcttacg tgccgatcaa cgtctcattt tcgccaaaag ttggcccagg
     2821 gcttcccggt atcaacaggg acaccaggat ttatttattc tgcgaagtga tcttccgtca
     2881 caggtattta ttcggcgcct gtagtgccat ttacccccat tcactgccag agccgtgagc
     2941 gcagcgaact gaatgtcacg aaaaagacag cgacttaggt gcctgatggt cggagacaaa
     3001 aggaatattc agcgatttgc ccgagcttgc gagggtgcta cttaagcctt tagggtttta
     3061 aggtctgttt tgtagaggag caaacagcgt ttgcgacatc cttttgtaat actgcggaac
     3121 tgactaaagt agtgagttat acacagggct gggatctatt ctttttatct ttttttattc
     3181 tttctttatt ctataaatta taaccacttg aatataaaca aaaaaaacac acaaaggtct
     3241 agcggaattt acagagggtc tagcagaatt tacaagtttt ccagcaaagg tctagcagaa
     3301 tttacagata cccacaactc aaaggaaaag gactagtaat tatcattgac tagcccatct
     3361 caattggtat agtgattaaa atcacctaga ccaattgaga tgtatgtctg aattagttgt
     3421 tttcaaagca aatgaactag cgattagtcg ctatgactta acggagcatg aaaccaagct
     3481 aattttatgc tgtgtggcac tactcaaccc cacgattgaa aaccctacaa ggaaagaacg
     3541 gacggtatcg ttcacttata accaatacgc tcagatgatg aacatcagta gggaaaatgc
     3601 ttatggtgta ttagctaaag caaccagaga gctgatgacg agaactgtgg aaatcaggaa
     3661 tcctttggtt aaaggctttg agattttcca gtggacaaac tatgccaagt tctcaagcga
     3721 aaaattagaa ttagttttta gtgaagagat attgccttat cttttccagt taaaaaaatt
     3781 cataaaatat aatctggaac atgttaagtc ttttgaaaac aaatactcta tgaggattta
     3841 tgagtggtta ttaaaagaac taacacaaaa gaaaactcac aaggcaaata tagagattag
     3901 ccttgatgaa tttaagttca tgttaatgct tgaaaataac taccatgagt ttaaaaggct
     3961 taaccaatgg gttttgaaac caataagtaa agatttaaac acttacagca atatgaaatt
     4021 ggtggttgat aagcgaggcc gcccgactga tacgttgatt ttccaagttg aactagatag
     4081 acaaatggat ctcgtaaccg aacttgagaa caaccagata aaaatgaatg gtgacaaaat
     4141 accaacaacc attacatcag attcctacct acataacgga ctaagaaaaa cactacacga
     4201 tgctttaact gcaaaaattc agctcaccag ttttgaggca aaatttttga gtgacatgca
     4261 aagtaagtat gatctcaatg gttcgttctc atggctcacg caaaaacaac gaaccacact
     4321 agagaacata ctggctaaat acggaaggat ctgaggttct tatggctctt gtatctatca
     4381 gtgaagcatc aagactaaca aacaaaagta gaacaactgt tcaccgttac atatcaaagg
     4441 gaaaactgtc catatgcaca gatgaaaacg gtgtaaaaaa gatagataca tcagagcttt
     4501 tacgagtttt tggtgcattt aaagctgttc accatgaaca gatcgacaat gtaacagatg
     4561 aacagcatgt aacacctaat agaacaggtg aaaccagtaa aacaaagcaa ctagaacatg
     4621 aaattgaaca cctgagacaa cttgttacag ctcaacagtc acacatagac agcctgaaac
     4681 aggcgatgct gcttatcgaa tcaaagctgc cgacaacacg ggagccagtg acgcctcccg
     4741 tggggaaaaa atcatggcaa ttctggaaga aatagcgccc aatacgcaaa ccgcctctcc
     4801 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     4861 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggcttaag
     4921 acccactttc acatttaagt tgtttttcta atccgcatat gatcaattca aggccgaata
     4981 agaaggctgg ctctgcacct tggtgatcaa ataattcgat agcttgtcgt aataatggcg
     5041 gcatactatc agtagtaggt gtttcccttt cttctttagc gacttgatgc tcttgatctt
     5101 ccaatacgca acctaaagta aaatgcccca cagcgctgag tgcatataat gcattctcta
     5161 gtgaaaaacc ttgttggcat aaaaaggcta attgattttc gagagtttca tactgttttt
     5221 ctgtaggccg tgtacctaaa tgtacttttg ctccatcgcg atgacttagt aaagcacatc
     5281 taaaactttt agcgttatta cgtaaaaaat cttgccagct ttccccttct aaagggcaaa
     5341 agtgagtatg gtgcctatct aacatctcaa tggctaaggc gtcgagcaaa gcccgcttat
     5401 tttttacatg ccaatacaat gtaggctgct ctacacctag cttctgggcg agtttacggg
     5461 ttgttaaacc ttcgattccg acctcattaa gcagctctaa tgcgctgtta atcactttac
     5521 ttttatctaa tctagacatc attaattcct aatttttggg tctactctat cattgatagc
     5581 gtgcctttac cactccctat catcgtt
//
