LOCUS       sequence-389812-        3109 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses truncated version of lambda CI named T3 under the control 
            of PLTetO-1 promoter, pUC origin of replication, Ampicillin 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3109)
  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 3109)
  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: pTA1cI-T3
FEATURES             Location/Qualifiers
     source          1..3109
                     /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             108..815
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="TKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQSG
                     VGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEYPV
                     FSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPDGM
                     LILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSVVG
                     KVIASQWPEETFG"
     terminator      827..913
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1014..1602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1094..1113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1920..2014)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2044..2835,2836..2904))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2667..2686
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2905..3009)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3077..3095
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagtccc tatcagtgat agagattgac atccctatca gtgatagaga tactgagcac
       61 atcagcagga cgcactgacc gaattcatta aagaggagaa aggtaccaca aaaaagaaac
      121 cattaacaca agagcagctt gaggacgcac gtcgccttaa agcaatttat gaaaaaaaga
      181 aaaatgaact tggcttatcc caggaatctg tcgcagacaa gatggggatg gggcagtcag
      241 gcgttggtgc tttatttaat ggcatcaatg cattaaatgc ttataacgcc gcattgcttg
      301 caaaaattct caaagttagc gttgaagaat ttagcccttc aatcgccaga gaaatctacg
      361 agatgtatga agcggttagt atgcagccgt cacttagaag tgagtatgag taccctgttt
      421 tttctcatgt tcaggcaggg atgttctcac ctgagcttag aacctttacc aaaggtgatg
      481 cggagagatg ggtaagcaca accaaaaaag ccagtgattc tgcattctgg cttgaggttg
      541 aaggtaattc catgaccgca ccaacaggct ccaagccaag ctttcctgac ggaatgttaa
      601 ttctcgttga ccctgagcag gctgttgagc caggtgattt ctgcatagcc agacttgggg
      661 gtgatgagtt taccttcaag aaactgatca gggatagcgg tcaggtgttt ttacaaccac
      721 taaacccaca gtacccaatg atcccatgca atgagagttg ttccgttgtg gggaaagtta
      781 tcgctagtca gtggcctgaa gagacgtttg gctgatctag aggcatcaaa taaaacgaaa
      841 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
      901 gagtaggaca aatccgccgc cctagaccta gtaggggata acgcaggaaa gaacatgtga
      961 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1021 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1081 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1141 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     1201 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     1261 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     1321 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     1381 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     1441 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     1501 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1561 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1621 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     1681 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     1741 taaagtatat atgagtaacc tgaggctatg gcagggcctg ccgccccgac gttggctgcg
     1801 agccctgggc cttcacccga acttgggggg tggggtgggg aaaaggaaga aacgcgggcg
     1861 tattggcccc aatggggtct cggtggggta tcgacagagt gccagcccct agtgcttgga
     1921 ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt
     1981 tctgaggtca ttactggatc tatcaacagg agtccaagcg agctcgtaaa cttggtctga
     2041 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2101 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     2161 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     2221 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     2281 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     2341 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     2401 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     2461 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     2521 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     2581 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     2641 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     2701 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     2761 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     2821 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     2881 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     2941 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3001 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     3061 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcac
//
