LOCUS       sequence-97746-e        6296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains PIq driving expression GalS-L, and PIq driving expression 
            of FruR-L..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6296)
  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 6296)
  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[GalS-L,FruR-L]
FEATURES             Location/Qualifiers
     source          1..6296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1054..1131)
                     /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     1175..1193
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1258..1335
                     /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    2615..2846
                     /gene="attP"
                     /label=lambda attP
                     /bound_moiety="lambda integrase"
                     /note="integrase from phage lambda"
     terminator      2881..2967
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3502..4452)
                     /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      4500..4722
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5214..5308)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(5338..6129,6130..6198))
                     /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     5961..5980
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6199..6290)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctcgagttac atcgcctgat ttgttgagtt agtgatcgcc gccgcattct ggcgtgttgc
       61 gacagaatgg cgacgcacta acgtcggcat aaaacagtgg ctggcacgag gatcaatatt
      121 gcctgctgcc ccctgcaagg ccagttcggt ggctaatttc gccattgaag caatgggata
      181 acgcacggtc gttaattgcg ggtcggtgta acgggcgatg ggaatatcat cgaacccgat
      241 gattgagaga tgtaacggaa tcgcaatgcc attatctttt aatgctgtca gtgcgccagc
      301 ggccatattg tcgttataag caaatacagc ggtaagttgt agattgcgcc ccagcagttc
      361 aaccatcgcc gcctcaccgc ccggcatgtc cggcgtacca gcgccaatcc agctttccgg
      421 cggaataata tcctgctctt tcaacgcact catccagcct gctttacgca tggcgtcatc
      481 ttcaatgccg tggctggaag aaagataacc aatacgttga tgaccgttat tcagcaacat
      541 gcgcgtcgcc attcgggcac cgctgagatt atccaggcaa acgcaacgat gggcgtaccc
      601 cggcacaacg cggttgatta acaccatacc gggaatgtta tccataaatt gcgccagttc
      661 atcgtcactc aatgcttttg agtgaacaat caacgcatta caacgctggc gaattaacac
      721 ctcaatggcg tgacgctctt tttccgcttc atgatagcta ttgccgatta gcacgtattt
      781 ctgatgctgc tgagcgacca gatccaccgc ttttaccagc gcgccgaaaa acgcatcaga
      841 aacgtccatc accaccacgc caatggtgtc gctctgtttg cccgccagtt gggctgccac
      901 gcggttggga atgtaattga gctccgccat cgccgcttcc actttttccc gcgttttcgc
      961 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     1021 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     1081 cgggcgctat catgccatac cgcgaaaggt tttgcaccat tcgatggtgt cgctagccta
     1141 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     1201 tcttcacctc gacaattcgc gcgcgaaggc gaagcggcat gcatttacgt tggtaccgac
     1261 accatcgaat ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa
     1321 ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc
     1381 tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg
     1441 gaaaaagtgg aagcggcgat ggcggagctc aattacattc ccaaccgcgt ggcacaacaa
     1501 ctggcgggca aaagtagccg ttctattggt cttgtgatcc ccgatctgga gaacaccagc
     1561 tatacccgca tcgctaacta tcttgaacgc caggcgcggc aacggggtta tcaactgctg
     1621 attgcctgct cagaagatca gccagacaac gaaatgcggt gcattgagca ccttttacag
     1681 cgtcaggttg atgccattat tgtttcgacg tcgttgcctc ctgagcatcc tttttatcaa
     1741 cgctgggcta acgacccgtt cccgattgtc gcgctggacc gcgccctcga tcgtgaacac
     1801 ttcaccagcg tggttggtgc cgatcaggat gatgccgaaa tgctggcgga agagttacgt
     1861 aagtttcccg ccgagacggt gctttatctt ggtgcgctac cggagctttc tgtcagcttc
     1921 ctgcgtgaac aaggtttccg tactgcctgg aaagatgatc cgcgcgaagt gcatttcctg
     1981 tatgccaaca gctatgagcg ggaggcggct gcccagttat tcgaaaaatg gctggaaacg
     2041 catccgatgc cgcaggcgct gttcacaacg tcgtttgcgt tgttgcaagg agtgatggat
     2101 gtcacgctgc gtcgcgacgg caaactgcct tctgacctgg caattgccac ctttggcgat
     2161 aacgaactgc tcgacttctt acagtgtccg gtgctggcag tggctcaacg tcaccgcgat
     2221 gtcgcagagc gtgtgctgga gattgtcctg gcaagcctgg acgaaccgcg taagccaaaa
     2281 cctggtttaa cgcgcattaa acgtaatctc tatcgccgcg gcgtgctcag ccgtagctaa
     2341 ccatgggcgc aacgcaatta atgtaagtta gcgcgaattg tcgagggaaa tcataaaaaa
     2401 tttatttgct ttcaggaaaa tttttctgta taatagattc ataaatttga gagaagagtt
     2461 taaatatggc tggttctcgc agaaagaaac atatccatga aatcccgccc cgaattcata
     2521 tgtctagata gctgagtagc ctagagtgcg cgacaggttt gatgacaaaa aattagcgca
     2581 agaagacaaa aatcaccttg cgctaatgct ctgttacagg tcactaatac catctaagta
     2641 gttgattcat agtgactgca tatgttgtgt tttacagtat tatgtagtct gttttttatg
     2701 caaaatctaa tttaatatat tgatatttat atcattttac gtttctcgtt cagctttttt
     2761 atactaagtt ggcattataa aaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2821 tatcagtcaa aataaaatca ttatttgatt tcaattttgt cccactcccc ctagaggcat
     2881 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     2941 gtgaacgctc tcctgagtag gacaaatccg ccgccctaga cctagggtac gggttttgct
     3001 gcccgcaaac gggctgttct ggtgttgcta gtttgttatc agaatcgcag atccggcttc
     3061 aggtttgccg gctgaaagcg ctatttcttc cagaattgcc atgatttttt ccccacggga
     3121 ggcgtcactg gctcccgtgt tgtcggcagc tttgattcga taagcagcat cgcctgtttc
     3181 aggctgtcta tgtgtgactg ttgagctgta acaagttgtc tcaggtgttc aatttcatgt
     3241 tctagttgct ttgttttact ggtttcacct gttctattag gtgttacatg ctgttcatct
     3301 gttacattgt cgatctgttc atggtgaaca gctttaaatg caccaaaaac tcgtaaaagc
     3361 tctgatgtat ctatcttttt tacaccgttt tcatctgtgc atatggacag ttttcccttt
     3421 gatatctaac ggtgaacagt tgttctactt ttgtttgtta gtcttgatgc ttcactgata
     3481 gatacaagag ccataagaac ctcagatcct tccgtattta gccagtatgt tctctagtgt
     3541 ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg agatcatgct tactttgcat
     3601 gtcactcaaa aattttgcct caaaactggt gagctgaatt tttgcagtta aagcatcgtg
     3661 tagtgttttt cttagtccgt tacgtaggta ggaatctgat gtaatggttg ttggtatttt
     3721 gtcaccattc atttttatct ggttgttctc aagttcggtt acgagatcca tttgtctatc
     3781 tagttcaact tggaaaatca acgtatcagt cgggcggcct cgcttatcaa ccaccaattt
     3841 catattgctg taagtgttta aatctttact tattggtttc aaaacccatt ggttaagcct
     3901 tttaaactca tggtagttat tttcaagcat taacatgaac ttaaattcat caaggctaat
     3961 ctctatattt gccttgtgag ttttcttttg tgttagttct tttaataacc actcataaat
     4021 cctcatagag tatttgtttt caaaagactt aacatgttcc agattatatt ttatgaattt
     4081 ttttaactgg aaaagataag gcaatatctc ttcactaaaa actaattcta atttttcgct
     4141 tgagaacttg gcatagtttg tccactggaa aatctcaaag cctttaacca aaggattcct
     4201 gatttccaca gttctcgtca tcagctctct ggttgcttta gctaatacac cataagcatt
     4261 ttccctactg atgttcatca tctgagcgta ttggttataa gtgaacgata ccgtccgttc
     4321 tttccttgta gggttttcaa tcgtggggtt gagtagtgcc acacagcata aaattagctt
     4381 ggtttcatgc tccgttaagt catagcgact aatcgctagt tcatttgctt tgaaaacaac
     4441 taattcagac atacatctca attggtctag gtgattttaa tcactatacc aattgagatg
     4501 ggctagtcaa tgataattac tagtcctttt cctttgagtt gtgggtatct gtaaattctg
     4561 ctagaccttt gctggaaaac ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc
     4621 tttgtgtgtt ttttttgttt atattcaagt ggttataatt tatagaataa agaaagaata
     4681 aaaaaagata aaaagaatag atcccagccc tgtgtataac tcactacttt agtcagttcc
     4741 gcagtattac aaaaggatgt cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa
     4801 ccctaaaggc ttaagtagca ccctcgcaag ctcgggcaaa tcgctgaata ttccttttgt
     4861 ctccgaccat caggcacctg agtcgctgtc tttttcgtga cattcagttc gctgcgctca
     4921 cggctctggc agtgaatggg ggtaaatggc actacaggcg ccttttatgg attcatgcaa
     4981 ggaaactacc cataatacaa gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg
     5041 gtctgctatg tggtgctatc tgactttttg ctgttcagca gttcctgccc tctgattttc
     5101 cagtctgacc acttcggatt atcccgtgac aggtcattca gactggctaa tgcacccagt
     5161 aaggcagcgg tatcatcaac aggcttaccc gtcttactgt ccctagtgct tggattctca
     5221 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag
     5281 gtcattactg gatctatcaa caggagtcca agcgagctcg taaacttggt ctgacagtta
     5341 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5401 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5461 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5521 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5581 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5641 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5701 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5761 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5821 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5881 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5941 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6001 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6061 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6121 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6181 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6241 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa tagggg
//
