LOCUS       sequence-97749-e        6330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains PIq driving expression RbsR-T, and PI driving expression of
            TreR-L..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6330)
  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 6330)
  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-T,LacI-T]
FEATURES             Location/Qualifiers
     source          1..6330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1000..1077)
                     /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     1121..1139
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1204..1281
                     /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."
     promoter        1204..1281
                     /gene="lacI"
                     /label=lacI promoter
     protein_bind    2606..2837
                     /gene="attP"
                     /label=lambda attP
                     /bound_moiety="lambda integrase"
                     /note="integrase from phage lambda"
     terminator      2872..2958
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3493..4443)
                     /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      4491..4713
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5205..5299)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(5329..6120,6121..6189))
                     /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     5952..5971
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6190..6281)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaagccgaa ccgcgttcca tcagaatcgg agtaagttgt aatcgttgtt gctgaagggt
       61 cggctgggtt atccgatgga tgagtacatc aatcgccagc tcccccagtt catctttcgg
      121 ttggtggata gtggttaatg gtggcgtcat aaagcttgcc agttcgatat cgtcatagcc
      181 aatcaccgcg atatcctgcg gaacctgtaa ctctgcctga tataacgcct ggtaaacgcc
      241 aacagccata gcgtcatttc cggtaaagac ggcctgagga cgcagcggat gtgatagcag
      301 ttggcgcata gcgtcaaacc cgccgttaaa ttcaaaatca ccagtgactt catagccatc
      361 aggaatgttg agacccgcac gtttcatcgc cgcccgataa ccttccaacc gcaggcgcgc
      421 cggagtttta tccagcgggc cggtaataca ggcgatacgg gtatgacctt tatcgatcag
      481 atattgcgtt gctaagtctc cgcccagcaa cgagttatcc tgaataagat cgctgtcgcc
      541 atcgaacgga gcccagtcca tcatcacagt aggcactgtc ggataacgtt gcatgatttc
      601 acgcgaaggt tgatgcgttt cggtgcacag taacagcaag ccatcaacgc gtttttgcat
      661 cagcgtttcc agattgcgat tcatccgctg ttcatcgcct tcggtattgc aaaggacgag
      721 actataaccg cgttcgaagc agctgcgttc aacgccacgc accagttctg aatagaaagg
      781 attggtactg gcagtgatca acatgccaat ggtatggctc gctttgcccg ccagttgttg
      841 tgccacgcgg ttgggaatgt aattgagctc cgccatcgcc gcttccactt tttcccgcgt
      901 tttcgcagaa acgtggctgg cctggttcac cacgttggaa acggtcgccg tagagacacc
      961 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     1021 ctcttccggg cgctatcatg ccataccgcg aaaggttttg caccattcga tggtgtcgct
     1081 agcctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     1141 ctttcgtctt cacctcgaca attcgcgcgc gaaggcgaag cggcatgcat ttacgttggt
     1201 accgacacca tcgaatgggg caaaaccttt cgcggtatgg catgatagcg cccggaagag
     1261 agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc
     1321 ggtgtctcta cggcgaccgt ttccaacgtg gtgaaccagg ccagccacgt ttctgcgaaa
     1381 acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca
     1441 caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg
     1501 cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc
     1561 gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat
     1621 cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc
     1681 attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag
     1741 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat
     1801 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg
     1861 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
     1921 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
     1981 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
     2041 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac
     2101 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
     2161 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
     2221 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat
     2281 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgtg accatgggcg
     2341 caacgcaatt aatgtaagtt agcgcgaatt gtcgagggaa atcataaaaa atttatttgc
     2401 tttcaggaaa atttttctgt ataatagatt cataaatttg agagaagagt ttaaatatgg
     2461 ctggttctcg cagaaagaaa catatccatg aaatcccgcc ccgaattcat atgtctagat
     2521 agctgagtag cctagagtgc gcgacaggtt tgatgacaaa aaattagcgc aagaagacaa
     2581 aaatcacctt gcgctaatgc tctgttacag gtcactaata ccatctaagt agttgattca
     2641 tagtgactgc atatgttgtg ttttacagta ttatgtagtc tgttttttat gcaaaatcta
     2701 atttaatata ttgatattta tatcatttta cgtttctcgt tcagcttttt tatactaagt
     2761 tggcattata aaaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     2821 aaataaaatc attatttgat ttcaattttg tcccactccc cctagaggca tcaaataaaa
     2881 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     2941 ctcctgagta ggacaaatcc gccgccctag acctagggta cgggttttgc tgcccgcaaa
     3001 cgggctgttc tggtgttgct agtttgttat cagaatcgca gatccggctt caggtttgcc
     3061 ggctgaaagc gctatttctt ccagaattgc catgattttt tccccacggg aggcgtcact
     3121 ggctcccgtg ttgtcggcag ctttgattcg ataagcagca tcgcctgttt caggctgtct
     3181 atgtgtgact gttgagctgt aacaagttgt ctcaggtgtt caatttcatg ttctagttgc
     3241 tttgttttac tggtttcacc tgttctatta ggtgttacat gctgttcatc tgttacattg
     3301 tcgatctgtt catggtgaac agctttaaat gcaccaaaaa ctcgtaaaag ctctgatgta
     3361 tctatctttt ttacaccgtt ttcatctgtg catatggaca gttttccctt tgatatctaa
     3421 cggtgaacag ttgttctact tttgtttgtt agtcttgatg cttcactgat agatacaaga
     3481 gccataagaa cctcagatcc ttccgtattt agccagtatg ttctctagtg tggttcgttg
     3541 tttttgcgtg agccatgaga acgaaccatt gagatcatgc ttactttgca tgtcactcaa
     3601 aaattttgcc tcaaaactgg tgagctgaat ttttgcagtt aaagcatcgt gtagtgtttt
     3661 tcttagtccg ttacgtaggt aggaatctga tgtaatggtt gttggtattt tgtcaccatt
     3721 catttttatc tggttgttct caagttcggt tacgagatcc atttgtctat ctagttcaac
     3781 ttggaaaatc aacgtatcag tcgggcggcc tcgcttatca accaccaatt tcatattgct
     3841 gtaagtgttt aaatctttac ttattggttt caaaacccat tggttaagcc ttttaaactc
     3901 atggtagtta ttttcaagca ttaacatgaa cttaaattca tcaaggctaa tctctatatt
     3961 tgccttgtga gttttctttt gtgttagttc ttttaataac cactcataaa tcctcataga
     4021 gtatttgttt tcaaaagact taacatgttc cagattatat tttatgaatt tttttaactg
     4081 gaaaagataa ggcaatatct cttcactaaa aactaattct aatttttcgc ttgagaactt
     4141 ggcatagttt gtccactgga aaatctcaaa gcctttaacc aaaggattcc tgatttccac
     4201 agttctcgtc atcagctctc tggttgcttt agctaataca ccataagcat tttccctact
     4261 gatgttcatc atctgagcgt attggttata agtgaacgat accgtccgtt ctttccttgt
     4321 agggttttca atcgtggggt tgagtagtgc cacacagcat aaaattagct tggtttcatg
     4381 ctccgttaag tcatagcgac taatcgctag ttcatttgct ttgaaaacaa ctaattcaga
     4441 catacatctc aattggtcta ggtgatttta atcactatac caattgagat gggctagtca
     4501 atgataatta ctagtccttt tcctttgagt tgtgggtatc tgtaaattct gctagacctt
     4561 tgctggaaaa cttgtaaatt ctgctagacc ctctgtaaat tccgctagac ctttgtgtgt
     4621 tttttttgtt tatattcaag tggttataat ttatagaata aagaaagaat aaaaaaagat
     4681 aaaaagaata gatcccagcc ctgtgtataa ctcactactt tagtcagttc cgcagtatta
     4741 caaaaggatg tcgcaaacgc tgtttgctcc tctacaaaac agaccttaaa accctaaagg
     4801 cttaagtagc accctcgcaa gctcgggcaa atcgctgaat attccttttg tctccgacca
     4861 tcaggcacct gagtcgctgt ctttttcgtg acattcagtt cgctgcgctc acggctctgg
     4921 cagtgaatgg gggtaaatgg cactacaggc gccttttatg gattcatgca aggaaactac
     4981 ccataataca agaaaagccc gtcacgggct tctcagggcg ttttatggcg ggtctgctat
     5041 gtggtgctat ctgacttttt gctgttcagc agttcctgcc ctctgatttt ccagtctgac
     5101 cacttcggat tatcccgtga caggtcattc agactggcta atgcacccag taaggcagcg
     5161 gtatcatcaa caggcttacc cgtcttactg tccctagtgc ttggattctc accaataaaa
     5221 aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact
     5281 ggatctatca acaggagtcc aagcgagctc gtaaacttgg tctgacagtt accaatgctt
     5341 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5401 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5461 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5521 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5581 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5641 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5701 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5761 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5821 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5881 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5941 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6001 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6061 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6121 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6181 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6241 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggctc gagttccgcg
     6301 cacatttccc cgaaaagtgc cacctgacgt
//
