LOCUS       sequence-389809-        2721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses frame-shifted cI under the control of PLTetO-1 promoter, 
            p15A origin of replication, Chloramphenicol selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2721)
  AUTHORS   Sarkar K, Mukhopadhyay S, Bonnerjee D, Srivastava R, Bagh S
  TITLE     A frame-shifted gene, which rescued its function by non-natural 
            start codons and its application in constructing synthetic gene 
            circuits.
  JOURNAL   J Biol Eng. 2019 Mar 1;13:20. doi: 10.1186/s13036-019-0151-x. 
            eCollection 2019.
  PUBMED    30867677
REFERENCE   2  (bases 1 to 2721)
  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: "J Biol 
            Eng."; date: "2019-03-1"; volume: "13:20. doi"; pages: " 
            10.1186/s13036-019-0151-x. eCollection 2019"
COMMENT     Sequence Label: pTC3cIfm
FEATURES             Location/Qualifiers
     source          1..2721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(7..25,26..31,32..48,49..54,55..80)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    7..25
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    32..50
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             90..101
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             113..820
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="TKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQSG
                     VGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEYPV
                     FSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPDGM
                     LILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSVVG
                     KVIASQWPEETFG"
     terminator      832..918
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(953..970)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1087..1632)
                     /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(1167..1186)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1746..1840)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1861..2520)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2435..2454
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2521..2623)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     2689..2707
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagtccc tatcagtgat agagattgac atccctatca gtgatagaga tactgagcac
       61 atcagcagga cgcactgacc gaattcatta aagaggagaa aggtaccatg gcacaaaaaa
      121 gaaaccatta acacaagagc agcttgagga cgcacgtcgc cttaaagcaa tttatgaaaa
      181 aaagaaaaat gaacttggct tatcccagga atctgtcgca gacaagatgg ggatggggca
      241 gtcaggcgtt ggtgctttat ttaatggcat caatgcatta aatgcttata acgccgcatt
      301 gcttgcaaaa attctcaaag ttagcgttga agaatttagc ccttcaatcg ccagagaaat
      361 ctacgagatg tatgaagcgg ttagtatgca gccgtcactt agaagtgagt atgagtaccc
      421 tgttttttct catgttcagg cagggatgtt ctcacctgag cttagaacct ttaccaaagg
      481 tgatgcggag agatgggtaa gcacaaccaa aaaagccagt gattctgcat tctggcttga
      541 ggttgaaggt aattccatga ccgcaccaac aggctccaag ccaagctttc ctgacggaat
      601 gttaattctc gttgaccctg agcaggctgt tgagccaggt gatttctgca tagccagact
      661 tgggggtgat gagtttacct tcaagaaact gatcagggat agcggtcagg tgtttttaca
      721 accactaaac ccacagtacc caatgatccc atgcaatgag agttgttccg ttgtggggaa
      781 agttatcgct agtcagtggc ctgaagagac gtttggctga tctagaggca tcaaataaaa
      841 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
      901 ctcctgagta ggacaaatcc gccgccctag acctagggat atattccgct tcctcgctca
      961 ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac gaacggggcg
     1021 gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg ccgcggcaaa
     1081 gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac gctcaaatca
     1141 gtggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gcggctccct
     1201 cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg
     1261 tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact
     1321 gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg
     1381 agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta
     1441 gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt
     1501 gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc
     1561 gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca
     1621 aaacgatctc aagaagatca tcttattaat cagataaaat atttctagat ttcagtgcaa
     1681 tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt gttactagtg
     1741 cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
     1801 tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa
     1861 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac
     1921 atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
     1981 gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc
     2041 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt
     2101 ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga
     2161 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt
     2221 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc
     2281 accgtctttc attgccatac ggaattccgg atgagcattc atcaggcggg caagaatgtg
     2341 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat
     2401 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg
     2461 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat
     2521 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     2581 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attttcgcca
     2641 gatatcgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     2701 acgaggccct ttcgtcttca c
//
