LOCUS       sequence-97754-e        6194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains PIq driving expression FruR-T, and PIq driving expression 
            of TreR-T..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6194)
  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 6194)
  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-T,TreR-T]
FEATURES             Location/Qualifiers
     source          1..6194
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1006..1083)
                     /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     1127..1145
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1210..1287
                     /gene="lacI"
                     /label=lacI promoter
     protein_bind    2507..2738
                     /gene="attP"
                     /label=lambda attP
                     /bound_moiety="lambda integrase"
                     /note="integrase from phage lambda"
     terminator      2773..2859
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3394..4344)
                     /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      4392..4614
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5106..5200)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(5230..6021,6022..6090))
                     /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     5853..5872
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6091..6182)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttagctacgg ctgagcacgc cgcggcgata gagattacgt ttaatgcgcg ttaaaccagg
       61 ttttggctta cgcggttcgt ccaggcttgc caggacaatc tccagcacac gctctgcgac
      121 atcgcggtga cgttgagcca ctgccagcac cggacactgt aagaagtcga gcagttcgtt
      181 atcgccaaag gtggcaattg ccaggtcaga aggcagtttg ccgtcgcgac gcagcgtgac
      241 atccatcact ccttgcaaca acgcaaacga cgttgtgaac agcgcctgcg gcatcggatg
      301 cgtttccagc catttttcga ataactgggc agccgcctcc cgctcatagc tgttggcata
      361 caggaaatgc acttcgcgcg gatcatcttt ccaggcagta cggaaacctt gttcacgcag
      421 gaagctgaca gaaagctccg gtagcgcacc aagataaagc accgtctcgg cgggaaactt
      481 acgtaactct tccgccagca tttcggcatc atcctgatcg gcaccaacca cgctggtgaa
      541 gtgttcacga tcgagggcgc ggtccagcgc gacaatcggg aacgggtcgt tagcccagcg
      601 ttgataaaaa ggatgctcag gaggcaacga cgtcgaaaca ataatggcat caacctgacg
      661 ctgtaaaagg tgctcaatgc accgcatttc gttgtctggc tgatcttctg agcaggcaat
      721 cagcagttga taaccccgtt gccgcgcctg gcgttcaaga tagttagcga tgcgggtata
      781 gctggtgttc tccagatcgg ggatcacaag accaatagaa cggctacttt tgcccgccag
      841 ttgttgtgcc acgcggttgg gaatgtaatt gagctccgcc atcgccgctt ccactttttc
      901 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg ttggaaacgg tcgccgtaga
      961 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     1021 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcacc attcgatggt
     1081 gtcgctagcc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     1141 gaggcccttt cgtcttcacc tcgacaattc gcgcgcgaag gcgaagcggc atgcatttac
     1201 gttggtaccg acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg
     1261 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
     1321 tatgccggtg tctctacggc gaccgtttcc aacgtggtga accaggccag ccacgtttct
     1381 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tcaattacat tcccaaccgc
     1441 gccgcacaac aactggcggg caaacagagc aaagtggtcg ccatcattgt tacccgtctg
     1501 gattcgttgt cagaaaatct cgccgttcaa accatgctgc cagcgttcta tgaacaaggt
     1561 tacgacccaa tcatgatgga aagtcagttt tccccgcaat tagttgccga acatttggga
     1621 gtgctgaaac ggcgtaatat cgacggcgta gtgctgttcg gttttactgg cataacagaa
     1681 gaaatgttag cccactggca gtcatcgctg gttctgctgg cgcgtgacgc aaaaggcttt
     1741 gcttcggtct gttatgacga cgaaggggca atcaaaatcc tgatgcaacg gctgtatgac
     1801 caggggcatc gtaatatcag ttatctcggc gtgccgcaca gtgacgtgac aaccggtaag
     1861 cgacgtcacg aagcctacct ggcgttctgc aaagcgcata aactgcatcc cgttgccgcc
     1921 ctgccagggc ttgctatgaa gcaaggctat gagaacgttg caaaagtgat tacgcctgaa
     1981 actaccgcct tactgtgcgc aaccgacacg ctggcacttg gcgcaagtaa atacctgcaa
     2041 gagcaacgca tcgacacctt gcaactggcg agcgtcggta atacgccgtt aatgaaattc
     2101 ctccatccgg agatcgtaac cgtagatccc ggttacgccg aagctggacg ccaggcggct
     2161 tgccagttga tcgcgcaggt aaccgggcgc agcgaaccgc aacaaatcat catccccgcc
     2221 accctgtcct gaccatgggc gcaacgcaat taatgtaagt tagcgcgaat tgtcgaggga
     2281 aatcataaaa aatttatttg ctttcaggaa aatttttctg tataatagat tcataaattt
     2341 gagagaagag tttaaatatg gctggttctc gcagaaagaa acatatccat gaaatcccgc
     2401 cccgaattca tatgtctaga tagctgagta gcctagagtg cgcgacaggt ttgatgacaa
     2461 aaaattagcg caagaagaca aaaatcacct tgcgctaatg ctctgttaca ggtcactaat
     2521 accatctaag tagttgattc atagtgactg catatgttgt gttttacagt attatgtagt
     2581 ctgtttttta tgcaaaatct aatttaatat attgatattt atatcatttt acgtttctcg
     2641 ttcagctttt ttatactaag ttggcattat aaaaaagcat tgcttatcaa tttgttgcaa
     2701 cgaacaggtc actatcagtc aaaataaaat cattatttga tttcaatttt gtcccactcc
     2761 ccctagaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     2821 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgcccta gacctagggt
     2881 acgggttttg ctgcccgcaa acgggctgtt ctggtgttgc tagtttgtta tcagaatcgc
     2941 agatccggct tcaggtttgc cggctgaaag cgctatttct tccagaattg ccatgatttt
     3001 ttccccacgg gaggcgtcac tggctcccgt gttgtcggca gctttgattc gataagcagc
     3061 atcgcctgtt tcaggctgtc tatgtgtgac tgttgagctg taacaagttg tctcaggtgt
     3121 tcaatttcat gttctagttg ctttgtttta ctggtttcac ctgttctatt aggtgttaca
     3181 tgctgttcat ctgttacatt gtcgatctgt tcatggtgaa cagctttaaa tgcaccaaaa
     3241 actcgtaaaa gctctgatgt atctatcttt tttacaccgt tttcatctgt gcatatggac
     3301 agttttccct ttgatatcta acggtgaaca gttgttctac ttttgtttgt tagtcttgat
     3361 gcttcactga tagatacaag agccataaga acctcagatc cttccgtatt tagccagtat
     3421 gttctctagt gtggttcgtt gtttttgcgt gagccatgag aacgaaccat tgagatcatg
     3481 cttactttgc atgtcactca aaaattttgc ctcaaaactg gtgagctgaa tttttgcagt
     3541 taaagcatcg tgtagtgttt ttcttagtcc gttacgtagg taggaatctg atgtaatggt
     3601 tgttggtatt ttgtcaccat tcatttttat ctggttgttc tcaagttcgg ttacgagatc
     3661 catttgtcta tctagttcaa cttggaaaat caacgtatca gtcgggcggc ctcgcttatc
     3721 aaccaccaat ttcatattgc tgtaagtgtt taaatcttta cttattggtt tcaaaaccca
     3781 ttggttaagc cttttaaact catggtagtt attttcaagc attaacatga acttaaattc
     3841 atcaaggcta atctctatat ttgccttgtg agttttcttt tgtgttagtt cttttaataa
     3901 ccactcataa atcctcatag agtatttgtt ttcaaaagac ttaacatgtt ccagattata
     3961 ttttatgaat ttttttaact ggaaaagata aggcaatatc tcttcactaa aaactaattc
     4021 taatttttcg cttgagaact tggcatagtt tgtccactgg aaaatctcaa agcctttaac
     4081 caaaggattc ctgatttcca cagttctcgt catcagctct ctggttgctt tagctaatac
     4141 accataagca ttttccctac tgatgttcat catctgagcg tattggttat aagtgaacga
     4201 taccgtccgt tctttccttg tagggttttc aatcgtgggg ttgagtagtg ccacacagca
     4261 taaaattagc ttggtttcat gctccgttaa gtcatagcga ctaatcgcta gttcatttgc
     4321 tttgaaaaca actaattcag acatacatct caattggtct aggtgatttt aatcactata
     4381 ccaattgaga tgggctagtc aatgataatt actagtcctt ttcctttgag ttgtgggtat
     4441 ctgtaaattc tgctagacct ttgctggaaa acttgtaaat tctgctagac cctctgtaaa
     4501 ttccgctaga cctttgtgtg ttttttttgt ttatattcaa gtggttataa tttatagaat
     4561 aaagaaagaa taaaaaaaga taaaaagaat agatcccagc cctgtgtata actcactact
     4621 ttagtcagtt ccgcagtatt acaaaaggat gtcgcaaacg ctgtttgctc ctctacaaaa
     4681 cagaccttaa aaccctaaag gcttaagtag caccctcgca agctcgggca aatcgctgaa
     4741 tattcctttt gtctccgacc atcaggcacc tgagtcgctg tctttttcgt gacattcagt
     4801 tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg gcactacagg cgccttttat
     4861 ggattcatgc aaggaaacta cccataatac aagaaaagcc cgtcacgggc ttctcagggc
     4921 gttttatggc gggtctgcta tgtggtgcta tctgactttt tgctgttcag cagttcctgc
     4981 cctctgattt tccagtctga ccacttcgga ttatcccgtg acaggtcatt cagactggct
     5041 aatgcaccca gtaaggcagc ggtatcatca acaggcttac ccgtcttact gtccctagtg
     5101 cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
     5161 tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct cgtaaacttg
     5221 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5281 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5341 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5401 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5461 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5521 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5581 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5641 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5701 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5761 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5821 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5881 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5941 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6001 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6061 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6121 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6181 aataggggct cgag
//
