LOCUS       sequence-97745-e        6293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains PIq driving expression GalS-L, and PIq driving expression 
            of RbsR-L..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6293)
  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 6293)
  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,RbsR-L]
FEATURES             Location/Qualifiers
     source          1..6293
                     /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    2612..2843
                     /gene="attP"
                     /label=lambda attP
                     /bound_moiety="lambda integrase"
                     /note="integrase from phage lambda"
     terminator      2878..2964
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3499..4449)
                     /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      4497..4719
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5211..5305)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(5335..6126,6127..6195))
                     /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     5958..5977
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6196..6287)
                     /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 aagcgagcca taccattggc atgttgatca ctgccagtac caatcctttc
     1561 tattcagaac tggtgcgtgg cgttgaacgc agctgcttcg aacgcggtta tagtctcgtc
     1621 ctttgcaata ccgaaggcga tgaacagcgg atgaatcgca atctggaaac gctgatgcaa
     1681 aaacgcgttg atggcttgct gttactgtgc accgaaacgc atcaaccttc gcgtgaaatc
     1741 atgcaacgtt atccgacagt gcctactgtg atgatggact gggctccgtt cgatggcgac
     1801 agcgatctta ttcaggataa ctcgttgctg ggcggagact tagcaacgca atatctgatc
     1861 gataaaggtc atacccgtat cgcctgtatt accggcccgc tggataaaac tccggcgcgc
     1921 ctgcggttgg aaggttatcg ggcggcgatg aaacgtgcgg gtctcaacat tcctgatggc
     1981 tatgaagtca ctggtgattt tgaatttaac ggcgggtttg acgctatgcg ccaactgcta
     2041 tcacatccgc tgcgtcctca ggccgtcttt accggaaatg acgctatggc tgttggcgtt
     2101 taccaggcgt tatatcaggc agagttacag gttccgcagg atatcgcggt gattggctat
     2161 gacgatatcg aactggcaag ctttatgacg ccaccattaa ccactatcca ccaaccgaaa
     2221 gatgaactgg gggagctggc gattgatgta ctcatccatc ggataaccca gccgaccctt
     2281 cagcaacaac gattacaact tactccgatt ctgatggaac gcggttcggc ttagtaacca
     2341 tgggcgcaac gcaattaatg taagttagcg cgaattgtcg agggaaatca taaaaaattt
     2401 atttgctttc aggaaaattt ttctgtataa tagattcata aatttgagag aagagtttaa
     2461 atatggctgg ttctcgcaga aagaaacata tccatgaaat cccgccccga attcatatgt
     2521 ctagatagct gagtagccta gagtgcgcga caggtttgat gacaaaaaat tagcgcaaga
     2581 agacaaaaat caccttgcgc taatgctctg ttacaggtca ctaataccat ctaagtagtt
     2641 gattcatagt gactgcatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa
     2701 aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag cttttttata
     2761 ctaagttggc attataaaaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     2821 cagtcaaaat aaaatcatta tttgatttca attttgtccc actcccccta gaggcatcaa
     2881 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     2941 aacgctctcc tgagtaggac aaatccgccg ccctagacct agggtacggg ttttgctgcc
     3001 cgcaaacggg ctgttctggt gttgctagtt tgttatcaga atcgcagatc cggcttcagg
     3061 tttgccggct gaaagcgcta tttcttccag aattgccatg attttttccc cacgggaggc
     3121 gtcactggct cccgtgttgt cggcagcttt gattcgataa gcagcatcgc ctgtttcagg
     3181 ctgtctatgt gtgactgttg agctgtaaca agttgtctca ggtgttcaat ttcatgttct
     3241 agttgctttg ttttactggt ttcacctgtt ctattaggtg ttacatgctg ttcatctgtt
     3301 acattgtcga tctgttcatg gtgaacagct ttaaatgcac caaaaactcg taaaagctct
     3361 gatgtatcta tcttttttac accgttttca tctgtgcata tggacagttt tccctttgat
     3421 atctaacggt gaacagttgt tctacttttg tttgttagtc ttgatgcttc actgatagat
     3481 acaagagcca taagaacctc agatccttcc gtatttagcc agtatgttct ctagtgtggt
     3541 tcgttgtttt tgcgtgagcc atgagaacga accattgaga tcatgcttac tttgcatgtc
     3601 actcaaaaat tttgcctcaa aactggtgag ctgaattttt gcagttaaag catcgtgtag
     3661 tgtttttctt agtccgttac gtaggtagga atctgatgta atggttgttg gtattttgtc
     3721 accattcatt tttatctggt tgttctcaag ttcggttacg agatccattt gtctatctag
     3781 ttcaacttgg aaaatcaacg tatcagtcgg gcggcctcgc ttatcaacca ccaatttcat
     3841 attgctgtaa gtgtttaaat ctttacttat tggtttcaaa acccattggt taagcctttt
     3901 aaactcatgg tagttatttt caagcattaa catgaactta aattcatcaa ggctaatctc
     3961 tatatttgcc ttgtgagttt tcttttgtgt tagttctttt aataaccact cataaatcct
     4021 catagagtat ttgttttcaa aagacttaac atgttccaga ttatatttta tgaatttttt
     4081 taactggaaa agataaggca atatctcttc actaaaaact aattctaatt tttcgcttga
     4141 gaacttggca tagtttgtcc actggaaaat ctcaaagcct ttaaccaaag gattcctgat
     4201 ttccacagtt ctcgtcatca gctctctggt tgctttagct aatacaccat aagcattttc
     4261 cctactgatg ttcatcatct gagcgtattg gttataagtg aacgataccg tccgttcttt
     4321 ccttgtaggg ttttcaatcg tggggttgag tagtgccaca cagcataaaa ttagcttggt
     4381 ttcatgctcc gttaagtcat agcgactaat cgctagttca tttgctttga aaacaactaa
     4441 ttcagacata catctcaatt ggtctaggtg attttaatca ctataccaat tgagatgggc
     4501 tagtcaatga taattactag tccttttcct ttgagttgtg ggtatctgta aattctgcta
     4561 gacctttgct ggaaaacttg taaattctgc tagaccctct gtaaattccg ctagaccttt
     4621 gtgtgttttt tttgtttata ttcaagtggt tataatttat agaataaaga aagaataaaa
     4681 aaagataaaa agaatagatc ccagccctgt gtataactca ctactttagt cagttccgca
     4741 gtattacaaa aggatgtcgc aaacgctgtt tgctcctcta caaaacagac cttaaaaccc
     4801 taaaggctta agtagcaccc tcgcaagctc gggcaaatcg ctgaatattc cttttgtctc
     4861 cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat tcagttcgct gcgctcacgg
     4921 ctctggcagt gaatgggggt aaatggcact acaggcgcct tttatggatt catgcaagga
     4981 aactacccat aatacaagaa aagcccgtca cgggcttctc agggcgtttt atggcgggtc
     5041 tgctatgtgg tgctatctga ctttttgctg ttcagcagtt cctgccctct gattttccag
     5101 tctgaccact tcggattatc ccgtgacagg tcattcagac tggctaatgc acccagtaag
     5161 gcagcggtat catcaacagg cttacccgtc ttactgtccc tagtgcttgg attctcacca
     5221 ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
     5281 attactggat ctatcaacag gagtccaagc gagctcgtaa acttggtctg acagttacca
     5341 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5401 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5461 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5521 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5581 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5641 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5701 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5761 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5821 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5881 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5941 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6001 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6061 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6121 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6181 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6241 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag ggg
//
