LOCUS       sequence-97748-e        6185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains PIq driving expression RbsR-L, and PI driving expression of
            TreR-L..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6185)
  AUTHORS   Shis DL, Hussain F, Meinhardt S, Swint-Kruse L, Bennett MR
  TITLE     Modular, Multi-Input Transcriptional Logic Gating with Orthogonal 
            LacI/GalR Family Chimeras.
  JOURNAL   ACS Synth Biol. 2014 Sep 19;3(9):645-51. doi: 10.1021/sb500262f. 
            Epub 2014 Jul 28.
  PUBMED    25035932
REFERENCE   2  (bases 1 to 6185)
  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"; doi: "10.1021/sb500262f";
            journalName: "ACS Synth Biol"; date: "2014-09-19- 19"; volume: "3"; 
            issue: "9"; pages: "645-51"
COMMENT     Sequence Label: pZS1[RbsR-L,TreR-L]
FEATURES             Location/Qualifiers
     source          1..6185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1009..1086)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     1130..1148
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1207..1284
                     /gene="lacI"
                     /label=lacI promoter
     protein_bind    2504..2735
                     /gene="attP"
                     /label=lambda attP
                     /bound_moiety="lambda integrase"
                     /note="integrase from phage lambda"
     terminator      2770..2856
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3391..4341)
                     /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      4389..4611
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5103..5197)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(5227..6018,6019..6087))
                     /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     5850..5869
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6088..6179)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctcgagttac taagccgaac cgcgttccat cagaatcgga gtaagttgta atcgttgttg
       61 ctgaagggtc ggctgggtta tccgatggat gagtacatca atcgccagct cccccagttc
      121 atctttcggt tggtggatag tggttaatgg tggcgtcata aagcttgcca gttcgatatc
      181 gtcatagcca atcaccgcga tatcctgcgg aacctgtaac tctgcctgat ataacgcctg
      241 gtaaacgcca acagccatag cgtcatttcc ggtaaagacg gcctgaggac gcagcggatg
      301 tgatagcagt tggcgcatag cgtcaaaccc gccgttaaat tcaaaatcac cagtgacttc
      361 atagccatca ggaatgttga gacccgcacg tttcatcgcc gcccgataac cttccaaccg
      421 caggcgcgcc ggagttttat ccagcgggcc ggtaatacag gcgatacggg tatgaccttt
      481 atcgatcaga tattgcgttg ctaagtctcc gcccagcaac gagttatcct gaataagatc
      541 gctgtcgcca tcgaacggag cccagtccat catcacagta ggcactgtcg gataacgttg
      601 catgatttca cgcgaaggtt gatgcgtttc ggtgcacagt aacagcaagc catcaacgcg
      661 tttttgcatc agcgtttcca gattgcgatt catccgctgt tcatcgcctt cggtattgca
      721 aaggacgaga ctataaccgc gttcgaagca gctgcgttca acgccacgca ccagttctga
      781 atagaaagga ttggtactgg cagtgatcaa catgccaatg gtatggctcg ctttgcccgc
      841 cagttgttgt gccacgcggt tgggaatgta attgagctcc gccatcgccg cttccacttt
      901 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
      961 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     1021 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc accattcgat
     1081 ggtgtcgcta gcctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     1141 cacgaggccc tttcgtcttc acctcgacaa ttcgcgcgcg aaggcgaagc ggcatgcatt
     1201 tacgttgaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa
     1261 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
     1321 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
     1381 aaaacgcggg aaaaagtgga agcggcgatg gcggagctca attacattcc caaccgcgcc
     1441 gcacaacaac tggcgggcaa acagagcaaa gtggtcgcca tcattgttac ccgtctggat
     1501 tcgttgtcag aaaatctcgc cgttcaaacc atgctgccag cgttctatga acaaggttac
     1561 gacccaatca tgatggaaag tcagttttcc ccgcaattag ttgccgaaca tttgggagtg
     1621 ctgaaacggc gtaatatcga cggcgtagtg ctgttcggtt ttactggcat aacagaagaa
     1681 atgttagccc actggcagtc atcgctggtt ctgctggcgc gtgacgcaaa aggctttgct
     1741 tcggtctgtt atgacgacga aggggcaatc aaaatcctga tgcaacggct gtatgaccag
     1801 gggcatcgta atatcagtta tctcggcgtg ccgcacagtg acgtgacaac cggtaagcga
     1861 cgtcacgaag cctacctggc gttctgcaaa gcgcataaac tgcatcccgt tgccgccctg
     1921 ccagggcttg ctatgaagca aggctatgag aacgttgcaa aagtgattac gcctgaaact
     1981 accgccttac tgtgcgcaac cgacacgctg gcacttggcg caagtaaata cctgcaagag
     2041 caacgcatcg acaccttgca actggcgagc gtcggtaata cgccgttaat gaaattcctc
     2101 catccggaga tcgtaaccgt agatcccggt tacgccgaag ctggacgcca ggcggcttgc
     2161 cagttgatcg cgcaggtaac cgggcgcagc gaaccgcaac aaatcatcat ccccgccacc
     2221 ctgtcctgac catgggcgca acgcaattaa tgtaagttag cgcgaattgt cgagggaaat
     2281 cataaaaaat ttatttgctt tcaggaaaat ttttctgtat aatagattca taaatttgag
     2341 agaagagttt aaatatggct ggttctcgca gaaagaaaca tatccatgaa atcccgcccc
     2401 gaattcatat gtctagatag ctgagtagcc tagagtgcgc gacaggtttg atgacaaaaa
     2461 attagcgcaa gaagacaaaa atcaccttgc gctaatgctc tgttacaggt cactaatacc
     2521 atctaagtag ttgattcata gtgactgcat atgttgtgtt ttacagtatt atgtagtctg
     2581 ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg tttctcgttc
     2641 agctttttta tactaagttg gcattataaa aaagcattgc ttatcaattt gttgcaacga
     2701 acaggtcact atcagtcaaa ataaaatcat tatttgattt caattttgtc ccactccccc
     2761 tagaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     2821 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgccctagac ctagggtacg
     2881 ggttttgctg cccgcaaacg ggctgttctg gtgttgctag tttgttatca gaatcgcaga
     2941 tccggcttca ggtttgccgg ctgaaagcgc tatttcttcc agaattgcca tgattttttc
     3001 cccacgggag gcgtcactgg ctcccgtgtt gtcggcagct ttgattcgat aagcagcatc
     3061 gcctgtttca ggctgtctat gtgtgactgt tgagctgtaa caagttgtct caggtgttca
     3121 atttcatgtt ctagttgctt tgttttactg gtttcacctg ttctattagg tgttacatgc
     3181 tgttcatctg ttacattgtc gatctgttca tggtgaacag ctttaaatgc accaaaaact
     3241 cgtaaaagct ctgatgtatc tatctttttt acaccgtttt catctgtgca tatggacagt
     3301 tttccctttg atatctaacg gtgaacagtt gttctacttt tgtttgttag tcttgatgct
     3361 tcactgatag atacaagagc cataagaacc tcagatcctt ccgtatttag ccagtatgtt
     3421 ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac gaaccattga gatcatgctt
     3481 actttgcatg tcactcaaaa attttgcctc aaaactggtg agctgaattt ttgcagttaa
     3541 agcatcgtgt agtgtttttc ttagtccgtt acgtaggtag gaatctgatg taatggttgt
     3601 tggtattttg tcaccattca tttttatctg gttgttctca agttcggtta cgagatccat
     3661 ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac
     3721 caccaatttc atattgctgt aagtgtttaa atctttactt attggtttca aaacccattg
     3781 gttaagcctt ttaaactcat ggtagttatt ttcaagcatt aacatgaact taaattcatc
     3841 aaggctaatc tctatatttg ccttgtgagt tttcttttgt gttagttctt ttaataacca
     3901 ctcataaatc ctcatagagt atttgttttc aaaagactta acatgttcca gattatattt
     3961 tatgaatttt tttaactgga aaagataagg caatatctct tcactaaaaa ctaattctaa
     4021 tttttcgctt gagaacttgg catagtttgt ccactggaaa atctcaaagc ctttaaccaa
     4081 aggattcctg atttccacag ttctcgtcat cagctctctg gttgctttag ctaatacacc
     4141 ataagcattt tccctactga tgttcatcat ctgagcgtat tggttataag tgaacgatac
     4201 cgtccgttct ttccttgtag ggttttcaat cgtggggttg agtagtgcca cacagcataa
     4261 aattagcttg gtttcatgct ccgttaagtc atagcgacta atcgctagtt catttgcttt
     4321 gaaaacaact aattcagaca tacatctcaa ttggtctagg tgattttaat cactatacca
     4381 attgagatgg gctagtcaat gataattact agtccttttc ctttgagttg tgggtatctg
     4441 taaattctgc tagacctttg ctggaaaact tgtaaattct gctagaccct ctgtaaattc
     4501 cgctagacct ttgtgtgttt tttttgttta tattcaagtg gttataattt atagaataaa
     4561 gaaagaataa aaaaagataa aaagaataga tcccagccct gtgtataact cactacttta
     4621 gtcagttccg cagtattaca aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag
     4681 accttaaaac cctaaaggct taagtagcac cctcgcaagc tcgggcaaat cgctgaatat
     4741 tccttttgtc tccgaccatc aggcacctga gtcgctgtct ttttcgtgac attcagttcg
     4801 ctgcgctcac ggctctggca gtgaatgggg gtaaatggca ctacaggcgc cttttatgga
     4861 ttcatgcaag gaaactaccc ataatacaag aaaagcccgt cacgggcttc tcagggcgtt
     4921 ttatggcggg tctgctatgt ggtgctatct gactttttgc tgttcagcag ttcctgccct
     4981 ctgattttcc agtctgacca cttcggatta tcccgtgaca ggtcattcag actggctaat
     5041 gcacccagta aggcagcggt atcatcaaca ggcttacccg tcttactgtc cctagtgctt
     5101 ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg
     5161 agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctcgt aaacttggtc
     5221 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5281 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5341 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5401 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5461 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5521 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5581 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5641 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5701 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5761 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5821 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5881 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5941 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6001 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6061 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6121 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6181 agggg
//
