LOCUS       sequence-202795-        5496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCKTRBS derivative carrying LmrR-BzdB1_nSP chimera, also known as 
            ChBz02.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5496)
  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 5496)
  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-LmrR-BzdB1_nSP
FEATURES             Location/Qualifiers
     source          1..5496
                     /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(1916..2575)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2490..2509
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2576..2678)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      3023..3245
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             3293..4243
                     /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    4769..4790
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(4805..5431)
                     /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     5336..5355
                     /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 cagaaatacc aaaagaaatg ttacgagccc aaaccaatgt aattttgctc
      181 aatgtcctaa aacaaggaga taattatgtt tacggtatta tcaaacaagt caaggaagcc
      241 tcgaatggag aaatggaact taatgaagcc acgctctata cgatctttaa aagacttgaa
      301 aaggatggga ttatcagttc ttattgggga gatgaaagtc aaggcgggcg tcgcaagtat
      361 taccgactgc agagcgcaaa taaaattcgt gtgggtttaa tgctgcctta caccggtacc
      421 tatgcgagcc tgggtaatgc cattaccaat ggctttaaac tggccgttga acaaggtggt
      481 ggtaaactgg gtggtcgcga gattgagtac ttcaccgtgg atgatgaaag cgatgccgca
      541 aaagcgccgg aaaatgccaa caaactgatt aaacgtgata atgtggatgt gctgattggc
      601 accgttcata gcggcgttgc actggcaatg acccgtgttg cacgtgaaac caaaacactg
      661 ctgatcgttc cgaatgcggg tgcagatgaa attaccggtc cgctgtgtag cccgaatatt
      721 tttcgtacaa gctttagcgc atggcagccg gcatatgcaa tgggtcaggt gatggccgaa
      781 aaaaaacata aaaatgtggt gaccctgagc tggaaatata gcttcggcga acagagcgtg
      841 gcaggtttca aggaagcctt tgaaaaaggc ggcggcaaag tggttaaaga aatgtatctg
      901 ccgtttccga acgtggaatt tcagccgtat ctgacggaaa ttgccgcact gaaaccggat
      961 gccgtgtttg tgttttttgc cggcggtggt gcagtgaaat ttgtgaaaga ttatgacgcc
     1021 gcaggtctga aaaaaaccat tccgctgtat ggtagcggtt ttctgaccga tggtaccctg
     1081 gaagcacagg gtgatagcgc agaaggtatt ctgaccaccc tgcattatgc agatggtctg
     1141 gatattgcaa aagataaagc ctttcgtacc gcctatgcca tggccttcaa aacccagccg
     1201 gatgtttata gcgttcaggg ttatgatgca gcacagcttt ttgcagcagg tttggcgggt
     1261 gttgccggtg atgttaccaa acgtgaagca ctgattgcag caatggaaaa agcaaccatt
     1321 gatagcccgc gtggtcgttt taccctgagt aaagcacata atccggtgca ggatatttac
     1381 ctgcggaaag tggaagccaa acagaataaa gtgatcgcag tggcagcaaa agccctggca
     1441 gatccggcac gtggttgtcg tatgtaagat ctctagagtg gaccttggtt gtcgtatgta
     1501 agatcctcta gagtggacct cgtggttgtc gtatgtaaga tcctctagag tggacctcgt
     1561 ggttgtcgta tgtaagatcc tctagagtgg acctgcaggc atgcaagctt gcggccgccc
     1621 gtttttatca ggctctggga ggcagaataa atgatcatat cgtcaattat tacctccacg
     1681 gggagagcct gagcaaactg gcctcaggca tttgagaagc acacggtcac actgcttccg
     1741 gtagtcaata aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct
     1801 gaaccgacga ccgggtcgaa tttgctttcg aatttctgcc attcatccgc ttattatcac
     1861 ttattcaggc gtagcaacca ggcgtttaag ggcaccaata actgccttaa aaaaattacg
     1921 ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga
     1981 agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt
     2041 gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt
     2101 ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa
     2161 taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata
     2221 tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag
     2281 tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt
     2341 ctttcattgc catacgaaat tccggatgag cattcatcag gcgggcaaga atgtgaataa
     2401 aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca
     2461 gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt
     2521 tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag
     2581 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
     2641 cattatggtg aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccaaaagt
     2701 tggcccaggg cttcccggta tcaacaggga caccaggatt tatttattct gcgaagtgat
     2761 cttccgtcac aggtatttat tcggcgcctg tagtgccatt tacccccatt cactgccaga
     2821 gccgtgagcg cagcgaactg aatgtcacga aaaagacagc gacttaggtg cctgatggtc
     2881 ggagacaaaa ggaatattca gcgatttgcc cgagcttgcg agggtgctac ttaagccttt
     2941 agggttttaa ggtctgtttt gtagaggagc aaacagcgtt tgcgacatcc ttttgtaata
     3001 ctgcggaact gactaaagta gtgagttata cacagggctg ggatctattc tttttatctt
     3061 tttttattct ttctttattc tataaattat aaccacttga atataaacaa aaaaaacaca
     3121 caaaggtcta gcggaattta cagagggtct agcagaattt acaagttttc cagcaaaggt
     3181 ctagcagaat ttacagatac ccacaactca aaggaaaagg actagtaatt atcattgact
     3241 agcccatctc aattggtata gtgattaaaa tcacctagac caattgagat gtatgtctga
     3301 attagttgtt ttcaaagcaa atgaactagc gattagtcgc tatgacttaa cggagcatga
     3361 aaccaagcta attttatgct gtgtggcact actcaacccc acgattgaaa accctacaag
     3421 gaaagaacgg acggtatcgt tcacttataa ccaatacgct cagatgatga acatcagtag
     3481 ggaaaatgct tatggtgtat tagctaaagc aaccagagag ctgatgacga gaactgtgga
     3541 aatcaggaat cctttggtta aaggctttga gattttccag tggacaaact atgccaagtt
     3601 ctcaagcgaa aaattagaat tagtttttag tgaagagata ttgccttatc ttttccagtt
     3661 aaaaaaattc ataaaatata atctggaaca tgttaagtct tttgaaaaca aatactctat
     3721 gaggatttat gagtggttat taaaagaact aacacaaaag aaaactcaca aggcaaatat
     3781 agagattagc cttgatgaat ttaagttcat gttaatgctt gaaaataact accatgagtt
     3841 taaaaggctt aaccaatggg ttttgaaacc aataagtaaa gatttaaaca cttacagcaa
     3901 tatgaaattg gtggttgata agcgaggccg cccgactgat acgttgattt tccaagttga
     3961 actagataga caaatggatc tcgtaaccga acttgagaac aaccagataa aaatgaatgg
     4021 tgacaaaata ccaacaacca ttacatcaga ttcctaccta cataacggac taagaaaaac
     4081 actacacgat gctttaactg caaaaattca gctcaccagt tttgaggcaa aatttttgag
     4141 tgacatgcaa agtaagtatg atctcaatgg ttcgttctca tggctcacgc aaaaacaacg
     4201 aaccacacta gagaacatac tggctaaata cggaaggatc tgaggttctt atggctcttg
     4261 tatctatcag tgaagcatca agactaacaa acaaaagtag aacaactgtt caccgttaca
     4321 tatcaaaggg aaaactgtcc atatgcacag atgaaaacgg tgtaaaaaag atagatacat
     4381 cagagctttt acgagttttt ggtgcattta aagctgttca ccatgaacag atcgacaatg
     4441 taacagatga acagcatgta acacctaata gaacaggtga aaccagtaaa acaaagcaac
     4501 tagaacatga aattgaacac ctgagacaac ttgttacagc tcaacagtca cacatagaca
     4561 gcctgaaaca ggcgatgctg cttatcgaat caaagctgcc gacaacacgg gagccagtga
     4621 cgcctcccgt ggggaaaaaa tcatggcaat tctggaagaa atagcgccca atacgcaaac
     4681 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     4741 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     4801 aggcttaaga cccactttca catttaagtt gtttttctaa tccgcatatg atcaattcaa
     4861 ggccgaataa gaaggctggc tctgcacctt ggtgatcaaa taattcgata gcttgtcgta
     4921 ataatggcgg catactatca gtagtaggtg tttccctttc ttctttagcg acttgatgct
     4981 cttgatcttc caatacgcaa cctaaagtaa aatgccccac agcgctgagt gcatataatg
     5041 cattctctag tgaaaaacct tgttggcata aaaaggctaa ttgattttcg agagtttcat
     5101 actgtttttc tgtaggccgt gtacctaaat gtacttttgc tccatcgcga tgacttagta
     5161 aagcacatct aaaactttta gcgttattac gtaaaaaatc ttgccagctt tccccttcta
     5221 aagggcaaaa gtgagtatgg tgcctatcta acatctcaat ggctaaggcg tcgagcaaag
     5281 cccgcttatt ttttacatgc caatacaatg taggctgctc tacacctagc ttctgggcga
     5341 gtttacgggt tgttaaacct tcgattccga cctcattaag cagctctaat gcgctgttaa
     5401 tcactttact tttatctaat ctagacatca ttaattccta atttttgggt ctactctatc
     5461 attgatagcg tgcctttacc actccctatc atcgtt
//
