LOCUS       sequence-12518-e        6664 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6664)
  AUTHORS   Elowitz MB, Leibler S
  TITLE     A synthetic oscillatory network of transcriptional regulators.
  JOURNAL   Nature. 2000 Jan 20. 403(6767):335-8.
  PUBMED    10659856
REFERENCE   2  (bases 1 to 6664)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2000 Jan 20. 403(6767):335-8."
COMMENT     Sequence Label: repressilator (pZS1-lTlrLLtCL)
FEATURES             Location/Qualifiers
     source          1..6664
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    8..24
                     /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    31..47
                     /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             89..100
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             107..727
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(180..199)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      775..861
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             968..979
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             997..2076
                     /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     complement(1016..1035)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      2141..2227
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(2258..2276,2277..2282,2283..2299,2300..2305,
                     2306..2331)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    2258..2276
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2283..2301
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             2341..2352
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2359..3069
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     terminator      3141..3227
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3762..4712)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      4760..4982
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5474..5568)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(5598..6389,6390..6458))
                     /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     6221..6240
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6459..6563)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6631..6649
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgagaattg tgagcggata acaattgaca ttgtgagcgg ataacaagat actgagcaca
       61 tcagcaggac gcactgaccg aattcattaa agaggagaaa ggtaccatgt ccagattaga
      121 taaaagtaaa gtgattaaca gcgcattaga gctgcttaat gaggtcggaa tcgaaggttt
      181 aacaacccgt aaactcgccc agaagctagg tgtagagcag cctacattgt attggcatgt
      241 aaaaaataag cgggctttgc tcgacgcctt agccattgag atgttagata ggcaccatac
      301 tcacttttgc cctttagaag gggaaagctg gcaagatttt ttacgtaata acgctaaaag
      361 ttttagatgt gctttactaa gtcatcgcga tggagcaaaa gtacatttag gtacacggcc
      421 tacagaaaaa cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc aacaaggttt
      481 ttcactagag aatgcattat atgcactcag cgctgtgggg cattttactt taggttgcgt
      541 attggaagat caagagcatc aagtcgctaa agaagaaagg gaaacaccta ctactgatag
      601 tatgccgcca ttattacgac aagctatcga attatttgat caccaaggtg cagagccagc
      661 cttcttattc ggccttgaat tgatcatatg cggattagaa aaacaactta aatgtgaaag
      721 tgggtctgca gcaaacgacg aaaactacgc tttagcagct taatctagag gcatcaaata
      781 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
      841 gctctcctga gtaggacaaa tccgccgccc tagacctagc tgcaggtcga ggataaatat
      901 ctaacaccgt gcgtgttgac tattttacct ctggcggtga taatggttgc atgtactaga
      961 attcattaaa gaggagaaag gtaccatatg gtgaatgtga aaccagtaac gttatacgat
     1021 gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc
     1081 cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt
     1141 cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc
     1201 tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat
     1261 caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa
     1321 gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg
     1381 gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt
     1441 gatgtctctg accagacacc catcaacagt attattttct cccatgaaga cggtacgcga
     1501 ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca
     1561 ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat
     1621 caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa
     1681 accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag
     1741 atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc
     1801 tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc gttaaccacc
     1861 atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct
     1921 cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc
     1981 accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     2041 ctggcacgac aggtttcccg actggaaagc gggcaggcag caaacgacga aaactacgct
     2101 ttagcagctt aaaagcttaa ttagctgagt ctagaggcat caaataaaac gaaaggctca
     2161 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     2221 gacaaatccg ccgccctaga cctagctgca ggtcgagtcc ctatcagtga tagagattga
     2281 catccctatc agtgatagag atactgagca catcagcagg acgcactgac cgaattcatt
     2341 aaagaggaga aaggtaccat gagcacaaaa aagaaaccat taacacaaga gcagcttgag
     2401 gacgcacgtc gccttaaagc aatttatgaa aaaaagaaaa atgaacttgg cttatcccag
     2461 gaatctgtcg cagacaagat ggggatgggg cagtcaggcg ttggtgcttt atttaatggc
     2521 atcaatgcat taaatgctta taacgccgca ttgcttgcaa aaattctcaa agttagcgtt
     2581 gaagaattta gcccttcaat cgccagagaa atctacgaga tgtatgaagc ggttagtatg
     2641 cagccgtcac ttagaagtga gtatgagtac cctgtttttt ctcatgttca ggcagggatg
     2701 ttctcacctg agcttagaac ctttaccaaa ggtgatgcgg agagatgggt aagcacaacc
     2761 aaaaaagcca gtgattctgc attctggctt gaggttgaag gtaattccat gaccgcacca
     2821 acaggctcca agccaagctt tcctgacgga atgttaattc tcgttgaccc tgagcaggct
     2881 gttgagccag gtgatttctg catagccaga cttgggggtg atgagtttac cttcaagaaa
     2941 ctgatcaggg atagcggtca ggtgttttta caaccactaa acccacagta cccaatgatc
     3001 ccatgcaatg agagttgttc cgttgtgggg aaagttatcg ctagtcagtg gcctgaagag
     3061 acgtttggcg cagcaaacga cgaaaactac gctttagcag cttgatctag aggatcccat
     3121 ggtacgcgtg ctagaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     3181 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgccctaga
     3241 cctagggtac gggttttgct gcccgcaaac gggctgttct ggtgttgcta gtttgttatc
     3301 agaatcgcag atccggcttc aggtttgccg gctgaaagcg ctatttcttc cagaattgcc
     3361 atgatttttt ccccacggga ggcgtcactg gctcccgtgt tgtcggcagc tttgattcga
     3421 taagcagcat cgcctgtttc aggctgtcta tgtgtgactg ttgagctgta acaagttgtc
     3481 tcaggtgttc aatttcatgt tctagttgct ttgttttact ggtttcacct gttctattag
     3541 gtgttacatg ctgttcatct gttacattgt cgatctgttc atggtgaaca gctttaaatg
     3601 caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt tacaccgttt tcatctgtgc
     3661 atatggacag ttttcccttt gatatctaac ggtgaacagt tgttctactt ttgtttgtta
     3721 gtcttgatgc ttcactgata gatacaagag ccataagaac ctcagatcct tccgtattta
     3781 gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg
     3841 agatcatgct tactttgcat gtcactcaaa aattttgcct caaaactggt gagctgaatt
     3901 tttgcagtta aagcatcgtg tagtgttttt cttagtccgt tacgtaggta ggaatctgat
     3961 gtaatggttg ttggtatttt gtcaccattc atttttatct ggttgttctc aagttcggtt
     4021 acgagatcca tttgtctatc tagttcaact tggaaaatca acgtatcagt cgggcggcct
     4081 cgcttatcaa ccaccaattt catattgctg taagtgttta aatctttact tattggtttc
     4141 aaaacccatt ggttaagcct tttaaactca tggtagttat tttcaagcat taacatgaac
     4201 ttaaattcat caaggctaat ctctatattt gccttgtgag ttttcttttg tgttagttct
     4261 tttaataacc actcataaat cctcatagag tatttgtttt caaaagactt aacatgttcc
     4321 agattatatt ttatgaattt ttttaactgg aaaagataag gcaatatctc ttcactaaaa
     4381 actaattcta atttttcgct tgagaacttg gcatagtttg tccactggaa aatctcaaag
     4441 cctttaacca aaggattcct gatttccaca gttctcgtca tcagctctct ggttgcttta
     4501 gctaatacac cataagcatt ttccctactg atgttcatca tctgagcgta ttggttataa
     4561 gtgaacgata ccgtccgttc tttccttgta gggttttcaa tcgtggggtt gagtagtgcc
     4621 acacagcata aaattagctt ggtttcatgc tccgttaagt catagcgact aatcgctagt
     4681 tcatttgctt tgaaaacaac taattcagac atacatctca attggtctag gtgattttaa
     4741 tcactatacc aattgagatg ggctagtcaa tgataattac tagtcctttt cctttgagtt
     4801 gtgggtatct gtaaattctg ctagaccttt gctggaaaac ttgtaaattc tgctagaccc
     4861 tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt atattcaagt ggttataatt
     4921 tatagaataa agaaagaata aaaaaagata aaaagaatag atcccagccc tgtgtataac
     4981 tcactacttt agtcagttcc gcagtattac aaaaggatgt cgcaaacgct gtttgctcct
     5041 ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag ctcgggcaaa
     5101 tcgctgaata ttccttttgt ctccgaccat caggcacctg agtcgctgtc tttttcgtga
     5161 cattcagttc gctgcgctca cggctctggc agtgaatggg ggtaaatggc actacaggcg
     5221 ccttttatgg attcatgcaa ggaaactacc cataatacaa gaaaagcccg tcacgggctt
     5281 ctcagggcgt tttatggcgg gtctgctatg tggtgctatc tgactttttg ctgttcagca
     5341 gttcctgccc tctgattttc cagtctgacc acttcggatt atcccgtgac aggtcattca
     5401 gactggctaa tgcacccagt aaggcagcgg tatcatcaac aggcttaccc gtcttactgt
     5461 ccctagtgct tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca
     5521 aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg
     5581 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5641 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5701 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5761 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5821 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5881 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5941 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6001 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6061 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6121 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6181 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6241 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6301 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6361 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6421 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6481 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6541 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     6601 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt
     6661 cacc
//
