LOCUS       sequence-97743-e        6251 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains PIq driving expression TreR-L, and PIq driving expression 
            of RbsR-L.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6251)
  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 6251)
  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[FruR-L,RbsR-L]
FEATURES             Location/Qualifiers
     source          1..6251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1012..1089)
                     /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     1133..1151
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1216..1293
                     /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."
     protein_bind    2570..2801
                     /gene="attP"
                     /label=lambda attP
                     /bound_moiety="lambda integrase"
                     /note="integrase from phage lambda"
     terminator      2836..2922
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3457..4407)
                     /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      4455..4677
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5169..5263)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(5293..6084,6085..6153))
                     /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     5916..5935
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6154..6245)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctcgagttag ctacggctga gcacgccgcg gcgatagaga ttacgtttaa tgcgcgttaa
       61 accaggtttt ggcttacgcg gttcgtccag gcttgccagg acaatctcca gcacacgctc
      121 tgcgacatcg cggtgacgtt gagccactgc cagcaccgga cactgtaaga agtcgagcag
      181 ttcgttatcg ccaaaggtgg caattgccag gtcagaaggc agtttgccgt cgcgacgcag
      241 cgtgacatcc atcactcctt gcaacaacgc aaacgacgtt gtgaacagcg cctgcggcat
      301 cggatgcgtt tccagccatt tttcgaataa ctgggcagcc gcctcccgct catagctgtt
      361 ggcatacagg aaatgcactt cgcgcggatc atctttccag gcagtacgga aaccttgttc
      421 acgcaggaag ctgacagaaa gctccggtag cgcaccaaga taaagcaccg tctcggcggg
      481 aaacttacgt aactcttccg ccagcatttc ggcatcatcc tgatcggcac caaccacgct
      541 ggtgaagtgt tcacgatcga gggcgcggtc cagcgcgaca atcgggaacg ggtcgttagc
      601 ccagcgttga taaaaaggat gctcaggagg caacgacgtc gaaacaataa tggcatcaac
      661 ctgacgctgt aaaaggtgct caatgcaccg catttcgttg tctggctgat cttctgagca
      721 ggcaatcagc agttgataac cccgttgccg cgcctggcgt tcaagatagt tagcgatgcg
      781 ggtatagctg gtgttctcca gatcggggat cacaagacca atagaacggc tacttttgcc
      841 cgccagttgt tgtgccacgc ggttgggaat gtaattgagc tccgccatcg ccgcttccac
      901 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
      961 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     1021 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcaccattc
     1081 gatggtgtcg ctagcctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     1141 tatcacgagg ccctttcgtc ttcacctcga caattcgcgc gcgaaggcga agcggcatgc
     1201 atttacgttg gtaccgacac catcgaatgg tgcaaaacct ttcgcggtat ggcatgatag
     1261 cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc
     1321 gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac
     1381 gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctcaa ttacattccc
     1441 aaccgcgtgg cacaacaact ggcgggcaaa gcgagccata ccattggcat gttgatcact
     1501 gccagtacca atcctttcta ttcagaactg gtgcgtggcg ttgaacgcag ctgcttcgaa
     1561 cgcggttata gtctcgtcct ttgcaatacc gaaggcgatg aacagcggat gaatcgcaat
     1621 ctggaaacgc tgatgcaaaa acgcgttgat ggcttgctgt tactgtgcac cgaaacgcat
     1681 caaccttcgc gtgaaatcat gcaacgttat ccgacagtgc ctactgtgat gatggactgg
     1741 gctccgttcg atggcgacag cgatcttatt caggataact cgttgctggg cggagactta
     1801 gcaacgcaat atctgatcga taaaggtcat acccgtatcg cctgtattac cggcccgctg
     1861 gataaaactc cggcgcgcct gcggttggaa ggttatcggg cggcgatgaa acgtgcgggt
     1921 ctcaacattc ctgatggcta tgaagtcact ggtgattttg aatttaacgg cgggtttgac
     1981 gctatgcgcc aactgctatc acatccgctg cgtcctcagg ccgtctttac cggaaatgac
     2041 gctatggctg ttggcgttta ccaggcgtta tatcaggcag agttacaggt tccgcaggat
     2101 atcgcggtga ttggctatga cgatatcgaa ctggcaagct ttatgacgcc accattaacc
     2161 actatccacc aaccgaaaga tgaactgggg gagctggcga ttgatgtact catccatcgg
     2221 ataacccagc cgacccttca gcaacaacga ttacaactta ctccgattct gatggaacgc
     2281 ggttcggctt agtaaccatg ggcgcaacgc aattaatgta agttagcgcg aattgtcgag
     2341 ggaaatcata aaaaatttat ttgctttcag gaaaattttt ctgtataata gattcataaa
     2401 tttgagagaa gagtttaaat atggctggtt ctcgcagaaa gaaacatatc catgaaatcc
     2461 cgccccgaat tcatatgtct agatagctga gtagcctaga gtgcgcgaca ggtttgatga
     2521 caaaaaatta gcgcaagaag acaaaaatca ccttgcgcta atgctctgtt acaggtcact
     2581 aataccatct aagtagttga ttcatagtga ctgcatatgt tgtgttttac agtattatgt
     2641 agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc
     2701 tcgttcagct tttttatact aagttggcat tataaaaaag cattgcttat caatttgttg
     2761 caacgaacag gtcactatca gtcaaaataa aatcattatt tgatttcaat tttgtcccac
     2821 tccccctaga ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     2881 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccgcc ctagacctag
     2941 ggtacgggtt ttgctgcccg caaacgggct gttctggtgt tgctagtttg ttatcagaat
     3001 cgcagatccg gcttcaggtt tgccggctga aagcgctatt tcttccagaa ttgccatgat
     3061 tttttcccca cgggaggcgt cactggctcc cgtgttgtcg gcagctttga ttcgataagc
     3121 agcatcgcct gtttcaggct gtctatgtgt gactgttgag ctgtaacaag ttgtctcagg
     3181 tgttcaattt catgttctag ttgctttgtt ttactggttt cacctgttct attaggtgtt
     3241 acatgctgtt catctgttac attgtcgatc tgttcatggt gaacagcttt aaatgcacca
     3301 aaaactcgta aaagctctga tgtatctatc ttttttacac cgttttcatc tgtgcatatg
     3361 gacagttttc cctttgatat ctaacggtga acagttgttc tacttttgtt tgttagtctt
     3421 gatgcttcac tgatagatac aagagccata agaacctcag atccttccgt atttagccag
     3481 tatgttctct agtgtggttc gttgtttttg cgtgagccat gagaacgaac cattgagatc
     3541 atgcttactt tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc
     3601 agttaaagca tcgtgtagtg tttttcttag tccgttacgt aggtaggaat ctgatgtaat
     3661 ggttgttggt attttgtcac cattcatttt tatctggttg ttctcaagtt cggttacgag
     3721 atccatttgt ctatctagtt caacttggaa aatcaacgta tcagtcgggc ggcctcgctt
     3781 atcaaccacc aatttcatat tgctgtaagt gtttaaatct ttacttattg gtttcaaaac
     3841 ccattggtta agccttttaa actcatggta gttattttca agcattaaca tgaacttaaa
     3901 ttcatcaagg ctaatctcta tatttgcctt gtgagttttc ttttgtgtta gttcttttaa
     3961 taaccactca taaatcctca tagagtattt gttttcaaaa gacttaacat gttccagatt
     4021 atattttatg aattttttta actggaaaag ataaggcaat atctcttcac taaaaactaa
     4081 ttctaatttt tcgcttgaga acttggcata gtttgtccac tggaaaatct caaagccttt
     4141 aaccaaagga ttcctgattt ccacagttct cgtcatcagc tctctggttg ctttagctaa
     4201 tacaccataa gcattttccc tactgatgtt catcatctga gcgtattggt tataagtgaa
     4261 cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca
     4321 gcataaaatt agcttggttt catgctccgt taagtcatag cgactaatcg ctagttcatt
     4381 tgctttgaaa acaactaatt cagacataca tctcaattgg tctaggtgat tttaatcact
     4441 ataccaattg agatgggcta gtcaatgata attactagtc cttttccttt gagttgtggg
     4501 tatctgtaaa ttctgctaga cctttgctgg aaaacttgta aattctgcta gaccctctgt
     4561 aaattccgct agacctttgt gtgttttttt tgtttatatt caagtggtta taatttatag
     4621 aataaagaaa gaataaaaaa agataaaaag aatagatccc agccctgtgt ataactcact
     4681 actttagtca gttccgcagt attacaaaag gatgtcgcaa acgctgtttg ctcctctaca
     4741 aaacagacct taaaacccta aaggcttaag tagcaccctc gcaagctcgg gcaaatcgct
     4801 gaatattcct tttgtctccg accatcaggc acctgagtcg ctgtcttttt cgtgacattc
     4861 agttcgctgc gctcacggct ctggcagtga atgggggtaa atggcactac aggcgccttt
     4921 tatggattca tgcaaggaaa ctacccataa tacaagaaaa gcccgtcacg ggcttctcag
     4981 ggcgttttat ggcgggtctg ctatgtggtg ctatctgact ttttgctgtt cagcagttcc
     5041 tgccctctga ttttccagtc tgaccacttc ggattatccc gtgacaggtc attcagactg
     5101 gctaatgcac ccagtaaggc agcggtatca tcaacaggct tacccgtctt actgtcccta
     5161 gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
     5221 agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgtaaac
     5281 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5341 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5401 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5461 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5521 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5581 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5641 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5701 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5761 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5821 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5881 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5941 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6001 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6061 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6121 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6181 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6241 acaaataggg g
//
