LOCUS       sequence-389811-        2722 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses lambda 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 2722)
  AUTHORS   Sarkar K, Bonnerjee D, Srivastava R, Bagh S
  TITLE     A single layer artificial neural network type architecture with 
            molecular engineered bacteria for reversible and irreversible 
            computing.
  JOURNAL   Chem Sci. 2021 Nov 9;12(48):15821-15832. doi: 10.1039/d1sc01505b. 
            eCollection 2021 Dec 15.
  PUBMED    35024106
REFERENCE   2  (bases 1 to 2722)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1039/d1sc01505b"; journalName: "Chem Sci"; date: "2021-11-9- 9";
            volume: "12"; issue: "48"; pages: "15821-15832"
COMMENT     Sequence Label: pTC3RBSC1cI
FEATURES             Location/Qualifiers
     source          1..2722
                     /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"
     CDS             108..821
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     terminator      833..919
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(954..971)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1088..1633)
                     /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(1168..1187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1747..1841)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1862..2521)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2436..2455
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2522..2624)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     2690..2708
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagtccc tatcagtgat agagattgac atccctatca gtgatagaga tactgagcac
       61 atcagcagga cgcactgacc gaattcattt ttgaggagtt cggtaccatg agcacaaaaa
      121 agaaaccatt aacacaagag cagcttgagg acgcacgtcg ccttaaagca atttatgaaa
      181 aaaagaaaaa tgaacttggc ttatcccagg aatctgtcgc agacaagatg gggatggggc
      241 agtcaggcgt tggtgcttta tttaatggca tcaatgcatt aaatgcttat aacgccgcat
      301 tgcttgcaaa aattctcaaa gttagcgttg aagaatttag cccttcaatc gccagagaaa
      361 tctacgagat gtatgaagcg gttagtatgc agccgtcact tagaagtgag tatgagtacc
      421 ctgttttttc tcatgttcag gcagggatgt tctcacctga gcttagaacc tttaccaaag
      481 gtgatgcgga gagatgggta agcacaacca aaaaagccag tgattctgca ttctggcttg
      541 aggttgaagg taattccatg accgcaccaa caggctccaa gccaagcttt cctgacggaa
      601 tgttaattct cgttgaccct gagcaggctg ttgagccagg tgatttctgc atagccagac
      661 ttgggggtga tgagtttacc ttcaagaaac tgatcaggga tagcggtcag gtgtttttac
      721 aaccactaaa cccacagtac ccaatgatcc catgcaatga gagttgttcc gttgtgggga
      781 aagttatcgc tagtcagtgg cctgaagaga cgtttggctg atctagaggc atcaaataaa
      841 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
      901 tctcctgagt aggacaaatc cgccgcccta gacctaggga tatattccgc ttcctcgctc
      961 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
     1021 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
     1081 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
     1141 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
     1201 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     1261 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     1321 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     1381 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     1441 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     1501 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     1561 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     1621 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca
     1681 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgttactagt
     1741 gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag
     1801 atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc tcgatatcaa
     1861 attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga
     1921 catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt
     1981 cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg
     2041 ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat
     2101 tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg
     2161 aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg
     2221 tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct
     2281 caccgtcttt cattgccata cggaattccg gatgagcatt catcaggcgg gcaagaatgt
     2341 gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa
     2401 tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat
     2461 gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca
     2521 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc
     2581 ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaacgtct cattttcgcc
     2641 agatatcgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     2701 cacgaggccc tttcgtcttc ac
//
