LOCUS       sequence-97750-e        6299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains PIq driving expression FruR-T, and PI driving expression of
            LacI-T..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6299)
  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 6299)
  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,LacI-T]
FEATURES             Location/Qualifiers
     source          1..6299
                     /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"
                     /label=lacI promoter
     protein_bind    2618..2849
                     /gene="attP"
                     /label=lambda attP
                     /bound_moiety="lambda integrase"
                     /note="integrase from phage lambda"
     terminator      2884..2970
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3505..4455)
                     /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      4503..4725
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5217..5311)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(5341..6132,6133..6201))
                     /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     5964..5983
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6202..6293)
                     /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 accacgttgg aaacggtcgc
      961 cgtagagaca 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 cgcaaaacct ttcgcggtat ggcatgatag
     1261 cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc
     1321 gcagagtatg ccggtgtctc tacggcgacc gtttccaacg tggtgaacca ggccagccac
     1381 gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc
     1441 aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc
     1501 agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa
     1561 ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg
     1621 gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat
     1681 gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat
     1741 gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg
     1801 ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta
     1861 agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa
     1921 attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc
     1981 atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg
     2041 gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg
     2101 gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc
     2161 aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag
     2221 ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc
     2281 ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     2341 tgaccatggg cgcaacgcaa ttaatgtaag ttagcgcgaa ttgtcgaggg aaatcataaa
     2401 aaatttattt gctttcagga aaatttttct gtataataga ttcataaatt tgagagaaga
     2461 gtttaaatat ggctggttct cgcagaaaga aacatatcca tgaaatcccg ccccgaattc
     2521 atatgtctag atagctgagt agcctagagt gcgcgacagg tttgatgaca aaaaattagc
     2581 gcaagaagac aaaaatcacc ttgcgctaat gctctgttac aggtcactaa taccatctaa
     2641 gtagttgatt catagtgact gcatatgttg tgttttacag tattatgtag tctgtttttt
     2701 atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc gttcagcttt
     2761 tttatactaa gttggcatta taaaaaagca ttgcttatca atttgttgca acgaacaggt
     2821 cactatcagt caaaataaaa tcattatttg atttcaattt tgtcccactc cccctagagg
     2881 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     2941 tcggtgaacg ctctcctgag taggacaaat ccgccgccct agacctaggg tacgggtttt
     3001 gctgcccgca aacgggctgt tctggtgttg ctagtttgtt atcagaatcg cagatccggc
     3061 ttcaggtttg ccggctgaaa gcgctatttc ttccagaatt gccatgattt tttccccacg
     3121 ggaggcgtca ctggctcccg tgttgtcggc agctttgatt cgataagcag catcgcctgt
     3181 ttcaggctgt ctatgtgtga ctgttgagct gtaacaagtt gtctcaggtg ttcaatttca
     3241 tgttctagtt gctttgtttt actggtttca cctgttctat taggtgttac atgctgttca
     3301 tctgttacat tgtcgatctg ttcatggtga acagctttaa atgcaccaaa aactcgtaaa
     3361 agctctgatg tatctatctt ttttacaccg ttttcatctg tgcatatgga cagttttccc
     3421 tttgatatct aacggtgaac agttgttcta cttttgtttg ttagtcttga tgcttcactg
     3481 atagatacaa gagccataag aacctcagat ccttccgtat ttagccagta tgttctctag
     3541 tgtggttcgt tgtttttgcg tgagccatga gaacgaacca ttgagatcat gcttactttg
     3601 catgtcactc aaaaattttg cctcaaaact ggtgagctga atttttgcag ttaaagcatc
     3661 gtgtagtgtt tttcttagtc cgttacgtag gtaggaatct gatgtaatgg ttgttggtat
     3721 tttgtcacca ttcattttta tctggttgtt ctcaagttcg gttacgagat ccatttgtct
     3781 atctagttca acttggaaaa tcaacgtatc agtcgggcgg cctcgcttat caaccaccaa
     3841 tttcatattg ctgtaagtgt ttaaatcttt acttattggt ttcaaaaccc attggttaag
     3901 ccttttaaac tcatggtagt tattttcaag cattaacatg aacttaaatt catcaaggct
     3961 aatctctata tttgccttgt gagttttctt ttgtgttagt tcttttaata accactcata
     4021 aatcctcata gagtatttgt tttcaaaaga cttaacatgt tccagattat attttatgaa
     4081 tttttttaac tggaaaagat aaggcaatat ctcttcacta aaaactaatt ctaatttttc
     4141 gcttgagaac ttggcatagt ttgtccactg gaaaatctca aagcctttaa ccaaaggatt
     4201 cctgatttcc acagttctcg tcatcagctc tctggttgct ttagctaata caccataagc
     4261 attttcccta ctgatgttca tcatctgagc gtattggtta taagtgaacg ataccgtccg
     4321 ttctttcctt gtagggtttt caatcgtggg gttgagtagt gccacacagc ataaaattag
     4381 cttggtttca tgctccgtta agtcatagcg actaatcgct agttcatttg ctttgaaaac
     4441 aactaattca gacatacatc tcaattggtc taggtgattt taatcactat accaattgag
     4501 atgggctagt caatgataat tactagtcct tttcctttga gttgtgggta tctgtaaatt
     4561 ctgctagacc tttgctggaa aacttgtaaa ttctgctaga ccctctgtaa attccgctag
     4621 acctttgtgt gttttttttg tttatattca agtggttata atttatagaa taaagaaaga
     4681 ataaaaaaag ataaaaagaa tagatcccag ccctgtgtat aactcactac tttagtcagt
     4741 tccgcagtat tacaaaagga tgtcgcaaac gctgtttgct cctctacaaa acagacctta
     4801 aaaccctaaa ggcttaagta gcaccctcgc aagctcgggc aaatcgctga atattccttt
     4861 tgtctccgac catcaggcac ctgagtcgct gtctttttcg tgacattcag ttcgctgcgc
     4921 tcacggctct ggcagtgaat gggggtaaat ggcactacag gcgcctttta tggattcatg
     4981 caaggaaact acccataata caagaaaagc ccgtcacggg cttctcaggg cgttttatgg
     5041 cgggtctgct atgtggtgct atctgacttt ttgctgttca gcagttcctg ccctctgatt
     5101 ttccagtctg accacttcgg attatcccgt gacaggtcat tcagactggc taatgcaccc
     5161 agtaaggcag cggtatcatc aacaggctta cccgtcttac tgtccctagt gcttggattc
     5221 tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct
     5281 gaggtcatta ctggatctat caacaggagt ccaagcgagc tcgtaaactt ggtctgacag
     5341 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5401 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5461 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5521 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5581 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5641 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5701 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5761 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5821 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5881 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5941 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6001 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6061 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6121 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6181 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6241 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaatagggg
//
