LOCUS       sequence-12603-e        6806 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 6806)
  AUTHORS   Cox RS 3rd, Dunlop MJ, Elowitz MB
  TITLE     A synthetic three-color scaffold for monitoring genetic regulation 
            and noise.
  JOURNAL   J Biol Eng. 2010 Jul 21;4:10. doi: 10.1186/1754-1611-4-10.
  PUBMED    20646328
REFERENCE   2  (bases 1 to 6806)
  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: "2010-07-21"; volume: "4:10. doi"; pages: " 
            10.1186/1754-1611-4-10"
COMMENT     Sequence Label: pZS2-123
FEATURES             Location/Qualifiers
     source          1..6806
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(1013..1038,1039..1044,1045..1061,1062..1067,
                     1068..1086))
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1043..1061
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1068..1086
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1325..1341
                     /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    1348..1364
                     /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)."
     terminator      complement(2455..2482)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2498..2569)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             3303..3323
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3364..3380
                     /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    3394..3413
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     CDS             3686..4480
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3740..3759)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4350..4369
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      4511..4605
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      5097..5319
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             5367..6317
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 gcggccgcaa aaggaaaaga tccggcaaac aaaccaccgt tggtagcggt ggtttttttg
       61 tttggatcga caatcttcgt aagcgtcatc aataagcgta aaaaaaccgg gcaatgcccg
      121 gttttttaat gagaaatttt acctgtcgta gccgccacca tccggcaaag aagcatacaa
      181 ggcttttggc ttatagctac gtagcgcatt gcgtcgcagc acaatcccgg caccgatcaa
      241 gtcttcgcga tgattattat tatttataca gctcatccat gccatgcgtg atgccagcag
      301 ccgttacaaa ttccagcagc accatgtgat cgcgcttctc attaggatct ttgctcagct
      361 tagattgagt gctcagatag tgattgtccg gcagcagaac aggaccgtcg ccaatcgggg
      421 tattttgttg ataatggtca gccagttgca cagagccgtc ctcgatatta tggcgaattt
      481 taaaatgcgc cttaatacca ttcttttgtt tatcggcggt gatatacaca ttatcgctaa
      541 tcgcattata ttccagttta tggcccagga tattaccatc ttccttaaaa tcgatgccct
      601 tcagctcaat acgattcacc agagtgtcgc cttcaaactt cacctctgcg cgagtcttgt
      661 aattaccatc atctttaaag aaaatggtgc gctcctgaac gtaaccctct ggcatggcgc
      721 tcttaaaaaa atcgtgctgc ttcatgtggt ctggatagcg agcgaagcat tgaacacccc
      781 aggtcagggt ggtaaccagg gtaggccaag gcaccggcag cttacccgtc gtacaaatga
      841 acttcagggt cagcttaccg taggttgcgt cgccttcacc ctcgccgcta acgctgaact
      901 tatgaccgtt gacatcaccg tccagttcga ccaggatagg aaccacacca gtaaacagct
      961 cctcgccctt gctcattttt ttttcctcct tattttctcc aggaagatct tcggtcagtg
     1021 cgtcctgctg atgtgctcag tatctctatc actgataggg atgtcaatct ctatcactga
     1081 tagggagtcg acaaaaataa tgagaatcaa tagaacttcc gagaagttca gccgctaata
     1141 atcgccctgc tccattgtgc gccgcaataa aagtaccggc attacgggtg cattggcgcg
     1201 ccaaatgcgg ccatctgtgg gcaacctgtg cggtaagacc caaacttagt gtaataggta
     1261 tcctatgatt attttttcat ttgatgccaa aaagacaatg aacccccgct cgagcggtcg
     1321 agaattgtga gcggataaca attgacattg tgagcggata acaagatact gagcacatca
     1381 gcaggacgca ctgaccgcgg gatcccggtg cagaaaataa ggaggaaaaa aaaatgagca
     1441 aaggtgaaga actgttcacc ggcgttgtgc caattctggt tgagctggat ggtgacgtga
     1501 atggccacaa attttccgtg tctggtgaag gcgagggtga tgctacttat ggcaaactga
     1561 ctctgaaact gatctgtacc accggcaaac tgcctgttcc gtggccaact ctggtcacta
     1621 ctctgggtta cggcctgatg tgttttgcgc gttacccgga tcacatgaaa cagcatgact
     1681 tcttcaaatc tgccatgccg gaaggctatg tccaagaacg tacgatcttt ttcaaggacg
     1741 acggcaacta taaaacccgt gccgaagtta aattcgaggg tgacaccctg gttaaccgca
     1801 tcgaactgaa aggcattgac ttcaaagagg acggcaacat tctgggtcac aagctggaat
     1861 acaactacaa ctcccacaac gtttacatta ctgctgacaa gcagaaaaac ggcatcaaag
     1921 caaacttcaa gatccgtcac aacattgaag atggtggcgt acagctggca gatcactacc
     1981 agcagaacac tccaatcggt gatggcccag tactgctgcc agataaccat tacctgtcct
     2041 accagagcaa actgtctaaa gacccgaacg aaaaacgtga ccacatggta ctgctggaat
     2101 ttgttaccgc ggcaggcatt acccacggta tggacgaact gtataaataa tgcaggtcgt
     2161 ctcggatcga gaaggacacg gttaatacta ggcctgctgg ctggtaatcg ccagcaggcc
     2221 tttttatttg ggggagaggg aagtcatgaa aaaactaacc tttgaaattc gatctccacc
     2281 acatcagctc tgaagcaacg taaaaaaacc cgccccggcg ggttttttta tacccgtagt
     2341 atccccactt atctacaata gctgtcctta attaaggttg aataaataaa aacagccgtt
     2401 gccagaaaga ggcacggctg tttttatttt ctagtgagac cgggagcagt taaacgcaga
     2461 aaggcccacc cgaaggtgag ccagtgtgac tctagtagag agcgttcacc gacaaacaac
     2521 agataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggtta
     2581 ttattatttg tacagctcat ccatgccacc ggtagaatga cgaccctccg cgcgctcata
     2641 ttgctctacg atcgtataat cttcattatg agaggtgatg tccagtttaa tattcacatt
     2701 gtacgcgcca ggcagctgca caggtttctt ggctttgtac gtggttttca cttcagcgtc
     2761 ataatggccg ccatctttca gtttcaggcg ctgtttaatt tcgcctttca gcgcaccatc
     2821 ttccggatac atgcgttcgc tagacgcctc ccaacccatc gtctttttct gcatcaccgg
     2881 gccatcagat ggaaaattag taccacgcag tttaacttta tagatgaact cgccatcctg
     2941 cagggaggag tcctgagtga cggtcacgac accaccatct tcaaaattca ttacgcgttc
     3001 ccacttgaaa ccttccggaa aagacagctt cagatagtcc gggatatccg ctgggtgttt
     3061 aacatacgct ttagaaccgt acataaattg cgggctcagg atgtcccacg caaaaggcag
     3121 cgggccgcct ttagtcactt tcagtttggc ggtctgggtg ccttcatacg gacggccctc
     3181 gccttcgcct tcgatctcga actcgtgacc gttaacagaa ccctccatgt gaactttgaa
     3241 gcgcatgaac tctttaatga tagccatgtt atcctcctcg cccttggaaa ccatggcagg
     3301 tgctccttct taaagttaaa caaaattatt tctagatttt ctcaagccta ggtctgtgtg
     3361 aaattgttat ccgctcacaa ttgaatctat cataattgtg agcgctcaca attgtaaagg
     3421 ttagatccgc taatcttatg gataaaaatg ctatgttccc cccgggggga tatcaacagg
     3481 agtccaagcg accggtggtt gcatgtctag ctagctagaa caggactagc taatggtttc
     3541 ttagacgtcg gaattgccag ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt
     3601 aaactggatg gctttcttgc cgccaaggat ctgatggcgc aggggatcaa gatctgatca
     3661 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3721 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3781 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3841 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     3901 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3961 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4021 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4081 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4141 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4201 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4261 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4321 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4381 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4441 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgag
     4501 agctcgcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
     4561 ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag cactagggac
     4621 agtaagacgg gtaagcctgt tgatgatacc gctgccttac tgggtgcatt agccagtctg
     4681 aatgacctgt cacgggataa tccgaagtgg tcagactgga aaatcagagg gcaggaactg
     4741 ctgaacagca aaaagtcaga tagcaccaca tagcagaccc gccataaaac gccctgagaa
     4801 gcccgtgacg ggcttttctt gtattatggg tagtttcctt gcatgaatcc ataaaaggcg
     4861 cctgtagtgc catttacccc cattcactgc cagagccgtg agcgcagcga actgaatgtc
     4921 acgaaaaaga cagcgactca ggtgcctgat ggtcggagac aaaaggaata ttcagcgatt
     4981 tgcccgagct tgcgagggtg ctacttaagc ctttagggtt ttaaggtctg ttttgtagag
     5041 gagcaaacag cgtttgcgac atccttttgt aatactgcgg aactgactaa agtagtgagt
     5101 tatacacagg gctgggatct attcttttta tcttttttta ttctttcttt attctataaa
     5161 ttataaccac ttgaatataa acaaaaaaaa cacacaaagg tctagcggaa tttacagagg
     5221 gtctagcaga atttacaagt tttccagcaa aggtctagca gaatttacag atacccacaa
     5281 ctcaaaggaa aaggactagt aattatcatt gactagccca tctcaattgg tatagtgatt
     5341 aaaatcacct agaccaattg agatgtatgt ctgaattagt tgttttcaaa gcaaatgaac
     5401 tagcgattag tcgctatgac ttaacggagc atgaaaccaa gctaatttta tgctgtgtgg
     5461 cactactcaa ccccacgatt gaaaacccta caaggaaaga acggacggta tcgttcactt
     5521 ataaccaata cgctcagatg atgaacatca gtagggaaaa tgcttatggt gtattagcta
     5581 aagcaaccag agagctgatg acgagaactg tggaaatcag gaatcctttg gttaaaggct
     5641 ttgagatttt ccagtggaca aactatgcca agttctcaag cgaaaaatta gaattagttt
     5701 ttagtgaaga gatattgcct tatcttttcc agttaaaaaa attcataaaa tataatctgg
     5761 aacatgttaa gtcttttgaa aacaaatact ctatgaggat ttatgagtgg ttattaaaag
     5821 aactaacaca aaagaaaact cacaaggcaa atatagagat tagccttgat gaatttaagt
     5881 tcatgttaat gcttgaaaat aactaccatg agtttaaaag gcttaaccaa tgggttttga
     5941 aaccaataag taaagattta aacacttaca gcaatatgaa attggtggtt gataagcgag
     6001 gccgcccgac tgatacgttg attttccaag ttgaactaga tagacaaatg gatctcgtaa
     6061 ccgaacttga gaacaaccag ataaaaatga atggtgacaa aataccaaca accattacat
     6121 cagattccta cctacgtaac ggactaagaa aaacactaca cgatgcttta actgcaaaaa
     6181 ttcagctcac cagttttgag gcaaaatttt tgagtgacat gcaaagtaag catgatctca
     6241 atggttcgtt ctcatggctc acgcaaaaac aacgaaccac actagagaac atactggcta
     6301 aatacggaag gatctgaggt tcttatggct cttgtatcta tcagtgaagc atcaagacta
     6361 acaaacaaaa gtagaacaac tgttcaccgt tagatatcaa agggaaaact gtccatatgc
     6421 acagatgaaa acggtgtaaa aaagatagat acatcagagc ttttacgagt ttttggtgca
     6481 tttaaagctg ttcaccatga acagatcgac aatgtaacag atgaacagca tgtaacacct
     6541 aatagaacag gtgaaaccag taaaacaaag caactagaac atgaaattga acacctgaga
     6601 caacttgtta cagctcaaca gtcacacata gacagcctga aacaggcgat gctgcttatc
     6661 gaatcaaagc tgccgacaac acgggagcca gtgacgcctc ccgtggggaa aaaatcatgg
     6721 caattctgga agaaatagcg ctttcagccg gcaaacctga agccggatct gcgattctga
     6781 taacaaacta gcaacaccag aacagc
//
