LOCUS       sequence-97747-e        6236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains PIq driving expression GalS-L, and PI driving expression of
            TreR-L..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6236)
  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 6236)
  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,TreR-L]
FEATURES             Location/Qualifiers
     source          1..6236
                     /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"
                     /label=lacI promoter
     protein_bind    2555..2786
                     /gene="attP"
                     /label=lambda attP
                     /bound_moiety="lambda integrase"
                     /note="integrase from phage lambda"
     terminator      2821..2907
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3442..4392)
                     /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      4440..4662
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5154..5248)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(5278..6069,6070..6138))
                     /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     5901..5920
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6139..6230)
                     /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 ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa
     1321 ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc
     1381 tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg
     1441 gaaaaagtgg aagcggcgat ggcggagctc aattacattc ccaaccgcgc cgcacaacaa
     1501 ctggcgggca aacagagcaa agtggtcgcc atcattgtta cccgtctgga ttcgttgtca
     1561 gaaaatctcg ccgttcaaac catgctgcca gcgttctatg aacaaggtta cgacccaatc
     1621 atgatggaaa gtcagttttc cccgcaatta gttgccgaac atttgggagt gctgaaacgg
     1681 cgtaatatcg acggcgtagt gctgttcggt tttactggca taacagaaga aatgttagcc
     1741 cactggcagt catcgctggt tctgctggcg cgtgacgcaa aaggctttgc ttcggtctgt
     1801 tatgacgacg aaggggcaat caaaatcctg atgcaacggc tgtatgacca ggggcatcgt
     1861 aatatcagtt atctcggcgt gccgcacagt gacgtgacaa ccggtaagcg acgtcacgaa
     1921 gcctacctgg cgttctgcaa agcgcataaa ctgcatcccg ttgccgccct gccagggctt
     1981 gctatgaagc aaggctatga gaacgttgca aaagtgatta cgcctgaaac taccgcctta
     2041 ctgtgcgcaa ccgacacgct ggcacttggc gcaagtaaat acctgcaaga gcaacgcatc
     2101 gacaccttgc aactggcgag cgtcggtaat acgccgttaa tgaaattcct ccatccggag
     2161 atcgtaaccg tagatcccgg ttacgccgaa gctggacgcc aggcggcttg ccagttgatc
     2221 gcgcaggtaa ccgggcgcag cgaaccgcaa caaatcatca tccccgccac cctgtcctga
     2281 ccatgggcgc aacgcaatta atgtaagtta gcgcgaattg tcgagggaaa tcataaaaaa
     2341 tttatttgct ttcaggaaaa tttttctgta taatagattc ataaatttga gagaagagtt
     2401 taaatatggc tggttctcgc agaaagaaac atatccatga aatcccgccc cgaattcata
     2461 tgtctagata gctgagtagc ctagagtgcg cgacaggttt gatgacaaaa aattagcgca
     2521 agaagacaaa aatcaccttg cgctaatgct ctgttacagg tcactaatac catctaagta
     2581 gttgattcat agtgactgca tatgttgtgt tttacagtat tatgtagtct gttttttatg
     2641 caaaatctaa tttaatatat tgatatttat atcattttac gtttctcgtt cagctttttt
     2701 atactaagtt ggcattataa aaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2761 tatcagtcaa aataaaatca ttatttgatt tcaattttgt cccactcccc ctagaggcat
     2821 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     2881 gtgaacgctc tcctgagtag gacaaatccg ccgccctaga cctagggtac gggttttgct
     2941 gcccgcaaac gggctgttct ggtgttgcta gtttgttatc agaatcgcag atccggcttc
     3001 aggtttgccg gctgaaagcg ctatttcttc cagaattgcc atgatttttt ccccacggga
     3061 ggcgtcactg gctcccgtgt tgtcggcagc tttgattcga taagcagcat cgcctgtttc
     3121 aggctgtcta tgtgtgactg ttgagctgta acaagttgtc tcaggtgttc aatttcatgt
     3181 tctagttgct ttgttttact ggtttcacct gttctattag gtgttacatg ctgttcatct
     3241 gttacattgt cgatctgttc atggtgaaca gctttaaatg caccaaaaac tcgtaaaagc
     3301 tctgatgtat ctatcttttt tacaccgttt tcatctgtgc atatggacag ttttcccttt
     3361 gatatctaac ggtgaacagt tgttctactt ttgtttgtta gtcttgatgc ttcactgata
     3421 gatacaagag ccataagaac ctcagatcct tccgtattta gccagtatgt tctctagtgt
     3481 ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg agatcatgct tactttgcat
     3541 gtcactcaaa aattttgcct caaaactggt gagctgaatt tttgcagtta aagcatcgtg
     3601 tagtgttttt cttagtccgt tacgtaggta ggaatctgat gtaatggttg ttggtatttt
     3661 gtcaccattc atttttatct ggttgttctc aagttcggtt acgagatcca tttgtctatc
     3721 tagttcaact tggaaaatca acgtatcagt cgggcggcct cgcttatcaa ccaccaattt
     3781 catattgctg taagtgttta aatctttact tattggtttc aaaacccatt ggttaagcct
     3841 tttaaactca tggtagttat tttcaagcat taacatgaac ttaaattcat caaggctaat
     3901 ctctatattt gccttgtgag ttttcttttg tgttagttct tttaataacc actcataaat
     3961 cctcatagag tatttgtttt caaaagactt aacatgttcc agattatatt ttatgaattt
     4021 ttttaactgg aaaagataag gcaatatctc ttcactaaaa actaattcta atttttcgct
     4081 tgagaacttg gcatagtttg tccactggaa aatctcaaag cctttaacca aaggattcct
     4141 gatttccaca gttctcgtca tcagctctct ggttgcttta gctaatacac cataagcatt
     4201 ttccctactg atgttcatca tctgagcgta ttggttataa gtgaacgata ccgtccgttc
     4261 tttccttgta gggttttcaa tcgtggggtt gagtagtgcc acacagcata aaattagctt
     4321 ggtttcatgc tccgttaagt catagcgact aatcgctagt tcatttgctt tgaaaacaac
     4381 taattcagac atacatctca attggtctag gtgattttaa tcactatacc aattgagatg
     4441 ggctagtcaa tgataattac tagtcctttt cctttgagtt gtgggtatct gtaaattctg
     4501 ctagaccttt gctggaaaac ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc
     4561 tttgtgtgtt ttttttgttt atattcaagt ggttataatt tatagaataa agaaagaata
     4621 aaaaaagata aaaagaatag atcccagccc tgtgtataac tcactacttt agtcagttcc
     4681 gcagtattac aaaaggatgt cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa
     4741 ccctaaaggc ttaagtagca ccctcgcaag ctcgggcaaa tcgctgaata ttccttttgt
     4801 ctccgaccat caggcacctg agtcgctgtc tttttcgtga cattcagttc gctgcgctca
     4861 cggctctggc agtgaatggg ggtaaatggc actacaggcg ccttttatgg attcatgcaa
     4921 ggaaactacc cataatacaa gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg
     4981 gtctgctatg tggtgctatc tgactttttg ctgttcagca gttcctgccc tctgattttc
     5041 cagtctgacc acttcggatt atcccgtgac aggtcattca gactggctaa tgcacccagt
     5101 aaggcagcgg tatcatcaac aggcttaccc gtcttactgt ccctagtgct tggattctca
     5161 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag
     5221 gtcattactg gatctatcaa caggagtcca agcgagctcg taaacttggt ctgacagtta
     5281 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5341 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5401 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5461 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5521 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5581 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5641 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5701 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5761 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5821 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5881 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5941 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6001 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6061 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6121 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6181 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa tagggg
//
