LOCUS       sequence-51015-e        4282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4282)
  AUTHORS   Huang DC, Holtz WJ, Maharbiz MM
  TITLE     A genetic bistable switch utilizing nonlinear protein degradation.
  JOURNAL   J Biol Eng. 2012 Jul 9;6(1):9.
  PUBMED    22776405
REFERENCE   2  (bases 1 to 4282)
  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: "2012-07-9"; volume: "6(1)"; pages: "9"
COMMENT     Sequence Label: PLlacO-1-CImf
FEATURES             Location/Qualifiers
     source          1..4282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(49..1131)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1093..1112
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1132..1209)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1415..1434
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     protein_bind    1502..1518
                     /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    1525..1541
                     /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)."
     CDS             1604..2314
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     terminator      2434..2505
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2521..2548
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2706..3294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2786..2805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3382..3476)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3497..4156)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4071..4090
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4157..4259)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 gacgtcggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
       61 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      121 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
      181 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
      241 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
      301 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt
      361 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
      421 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
      481 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
      541 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
      601 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
      661 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
      721 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
      781 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
      841 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
      901 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
      961 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     1021 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     1081 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     1141 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     1201 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     1261 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     1321 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     1381 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     1441 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     1501 attgtgagcg gataacaatt gacattgtga gcggataaca agatactgag cacgaattca
     1561 aaagatctaa tccaacaata tagcatactc tcgcgacggc ctcatgagca caaaaaagaa
     1621 accattaaca caagagcagc ttgaggacgc acgtcgcctt aaagcaattt atgaaaaaaa
     1681 gaaaaatgaa cttggcttat cccaggaatc tgtcgcagac aagatgggga tggggcagtc
     1741 aggcgttggt gctttattta atggcatcaa tgcattaaat gcttataacg ccgcattgct
     1801 tgcaaaaatt ctcaaagtta gcgttgaaga atttagccct tcaatcgcca gagaaatcta
     1861 cgagatgtat gaagcggtta gtatgcagcc gtcacttaga agtgagtatg agtaccctgt
     1921 tttttctcat gttcaggcag ggatgttctc acctgagctt agaaccttta ccaaaggtga
     1981 tgcggagaga tgggtaagca caaccaaaaa agccagtgat tctgcattct ggcttgaggt
     2041 tgaaggtaat tccatgaccg caccaacagg ctccaagcca agctttcctg acggaatgtt
     2101 aattctcgtt gaccctgagc aggctgttga gccaggtgat ttctgcatag ccagacttgg
     2161 gggtgatgag tttaccttca agaaactgat cagggatagc ggtcaggtgt ttttacaacc
     2221 actaaaccca cagtacccaa tgatcccatg caatgagagt tgttccgttg tggggaaagt
     2281 tatcgctagt cagtggcctg aagagacgtt tggcgcggcg aataaaaatg aagaaaatac
     2341 caatgaagtg ccgaccttta tgctgaatgc gggccaggcg aattatgcgt ttgcgtaata
     2401 aggatccaaa ctcgagtaag gatctccagg catcaaataa aacgaaaggc tcagtcgaaa
     2461 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact
     2521 ggctcacctt cgggtgggcc tttctgcgtt tatacctagg gcgttcggct gcggcgagcg
     2581 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2641 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2701 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2761 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2821 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2881 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2941 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3001 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3061 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3121 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     3181 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3241 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3301 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3361 ttggtcatga ctagtgcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt
     3421 tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag
     3481 cgagctcgat atcaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat
     3541 taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg
     3601 gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga
     3661 agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg
     3721 agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac
     3781 acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc
     3841 agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat
     3901 cccatatcac cagctcaccg tctttcattg ccatacgaaa ttccggatga gcattcatca
     3961 ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct
     4021 ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact
     4081 gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag
     4141 tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     4201 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     4261 cgtctcattt tcgccagata tc
//
