LOCUS       sequence-389837-        3466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses frame-shifted CI under the control of PLTATAA synthetic 
            promoter, carries a synthetic fragment with additional restriction 
            sites, ColE1 origin of replication, Ampicillin selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3466)
  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 3466)
  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: pDA2ScIfm
FEATURES             Location/Qualifiers
     source          1..3466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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..48
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    62..80
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    86..102
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    110..126
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             138..149
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             161..868
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="TKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQSG
                     VGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEYPV
                     FSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPDGM
                     LILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSVVG
                     KVIASQWPEETFG"
     terminator      880..966
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1131..1719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1211..1230)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1807..1901)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(1931..2722,2723..2791))
                     /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     2554..2573
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2792..2896)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2964..2982
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagtccc tatcagtgat agagattgac attgtgagcg gataacaaga tactgagcac
       61 atccctatca gtgatagaga gataattgtg agcggataac aattgataat tgtgagcgga
      121 taacaattga attcattaaa gaggagaaag gtaccatggc acaaaaaaga aaccattaac
      181 acaagagcag cttgaggacg cacgtcgcct taaagcaatt tatgaaaaaa agaaaaatga
      241 acttggctta tcccaggaat ctgtcgcaga caagatgggg atggggcagt caggcgttgg
      301 tgctttattt aatggcatca atgcattaaa tgcttataac gccgcattgc ttgcaaaaat
      361 tctcaaagtt agcgttgaag aatttagccc ttcaatcgcc agagaaatct acgagatgta
      421 tgaagcggtt agtatgcagc cgtcacttag aagtgagtat gagtaccctg ttttttctca
      481 tgttcaggca gggatgttct cacctgagct tagaaccttt accaaaggtg atgcggagag
      541 atgggtaagc acaaccaaaa aagccagtga ttctgcattc tggcttgagg ttgaaggtaa
      601 ttccatgacc gcaccaacag gctccaagcc aagctttcct gacggaatgt taattctcgt
      661 tgaccctgag caggctgttg agccaggtga tttctgcata gccagacttg ggggtgatga
      721 gtttaccttc aagaaactga tcagggatag cggtcaggtg tttttacaac cactaaaccc
      781 acagtaccca atgatcccat gcaatgagag ttgttccgtt gtggggaaag ttatcgctag
      841 tcagtggcct gaagagacgt ttggctgatc tagaggcatc aaataaaacg aaaggctcag
      901 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
      961 acaaatccgc cgccctagac ctagggcgtt cggctgcggc gagcggtatc agctcactca
     1021 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1081 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1141 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1201 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1261 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1321 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1381 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1441 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1501 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1561 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1621 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1681 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1741 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgactagt
     1801 gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag
     1861 atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc tcgtaaactt
     1921 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     1981 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     2041 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     2101 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     2161 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     2221 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     2281 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     2341 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     2401 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     2461 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     2521 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     2581 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     2641 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     2701 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     2761 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     2821 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     2881 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     2941 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcacttgt
     3001 tttggagcac ggaaagacga tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt
     3061 aacaaccgcc aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct
     3121 taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca agaaggatcc
     3181 tggcagaagc tatgaaattc tttatgccgg ctgcagcaga ttctcgcatg ccagagatcc
     3241 tatttttggc aatcaaatca ttccggatac tgcgatttta agtgttgttc cattccatca
     3301 cggttttgga atgtttacta cactcggata tttgatatgt ggatttcgag tcgtcttaat
     3361 gtatagattt gaagaagagc tgtttttacg atcccttcag gattacaaaa ttcaaagtgc
     3421 gttgctagta ccaaccctat tttcattctt cgccaaaagc actctg
//
