LOCUS       sequence-97739-e        6290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains PIq driving expression RbsR-L, and PI driving expression of
            LacI-L..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6290)
  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 6290)
  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[RbsR-L,LacI-L]
FEATURES             Location/Qualifiers
     source          1..6290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1009..1086)
                     /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     1130..1148
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1207..1284
                     /gene="lacI"
                     /label=lacI promoter
     protein_bind    2609..2840
                     /gene="attP"
                     /label=lambda attP
                     /bound_moiety="lambda integrase"
                     /note="integrase from phage lambda"
     terminator      2875..2961
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3496..4446)
                     /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      4494..4716
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5208..5302)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(5332..6123,6124..6192))
                     /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     5955..5974
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6193..6284)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctcgagttac taagccgaac cgcgttccat cagaatcgga gtaagttgta atcgttgttg
       61 ctgaagggtc ggctgggtta tccgatggat gagtacatca atcgccagct cccccagttc
      121 atctttcggt tggtggatag tggttaatgg tggcgtcata aagcttgcca gttcgatatc
      181 gtcatagcca atcaccgcga tatcctgcgg aacctgtaac tctgcctgat ataacgcctg
      241 gtaaacgcca acagccatag cgtcatttcc ggtaaagacg gcctgaggac gcagcggatg
      301 tgatagcagt tggcgcatag cgtcaaaccc gccgttaaat tcaaaatcac cagtgacttc
      361 atagccatca ggaatgttga gacccgcacg tttcatcgcc gcccgataac cttccaaccg
      421 caggcgcgcc ggagttttat ccagcgggcc ggtaatacag gcgatacggg tatgaccttt
      481 atcgatcaga tattgcgttg ctaagtctcc gcccagcaac gagttatcct gaataagatc
      541 gctgtcgcca tcgaacggag cccagtccat catcacagta ggcactgtcg gataacgttg
      601 catgatttca cgcgaaggtt gatgcgtttc ggtgcacagt aacagcaagc catcaacgcg
      661 tttttgcatc agcgtttcca gattgcgatt catccgctgt tcatcgcctt cggtattgca
      721 aaggacgaga ctataaccgc gttcgaagca gctgcgttca acgccacgca ccagttctga
      781 atagaaagga ttggtactgg cagtgatcaa catgccaatg gtatggctcg ctttgcccgc
      841 cagttgttgt gccacgcggt tgggaatgta attgagctcc gccatcgccg cttccacttt
      901 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
      961 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     1021 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc accattcgat
     1081 ggtgtcgcta gcctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     1141 cacgaggccc tttcgtcttc acctcgacaa ttcgcgcgcg aaggcgaagc ggcatgcatt
     1201 tacgttgaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa
     1261 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
     1321 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
     1381 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
     1441 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
     1501 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
     1561 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
     1621 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
     1681 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
     1741 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
     1801 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
     1861 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
     1921 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
     1981 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
     2041 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
     2101 tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat
     2161 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     2221 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     2281 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct gtgaccatgg
     2341 gcgcaacgca attaatgtaa gttagcgcga attgtcgagg gaaatcataa aaaatttatt
     2401 tgctttcagg aaaatttttc tgtataatag attcataaat ttgagagaag agtttaaata
     2461 tggctggttc tcgcagaaag aaacatatcc atgaaatccc gccccgaatt catatgtcta
     2521 gatagctgag tagcctagag tgcgcgacag gtttgatgac aaaaaattag cgcaagaaga
     2581 caaaaatcac cttgcgctaa tgctctgtta caggtcacta ataccatcta agtagttgat
     2641 tcatagtgac tgcatatgtt gtgttttaca gtattatgta gtctgttttt tatgcaaaat
     2701 ctaatttaat atattgatat ttatatcatt ttacgtttct cgttcagctt ttttatacta
     2761 agttggcatt ataaaaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     2821 tcaaaataaa atcattattt gatttcaatt ttgtcccact ccccctagag gcatcaaata
     2881 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     2941 gctctcctga gtaggacaaa tccgccgccc tagacctagg gtacgggttt tgctgcccgc
     3001 aaacgggctg ttctggtgtt gctagtttgt tatcagaatc gcagatccgg cttcaggttt
     3061 gccggctgaa agcgctattt cttccagaat tgccatgatt ttttccccac gggaggcgtc
     3121 actggctccc gtgttgtcgg cagctttgat tcgataagca gcatcgcctg tttcaggctg
     3181 tctatgtgtg actgttgagc tgtaacaagt tgtctcaggt gttcaatttc atgttctagt
     3241 tgctttgttt tactggtttc acctgttcta ttaggtgtta catgctgttc atctgttaca
     3301 ttgtcgatct gttcatggtg aacagcttta aatgcaccaa aaactcgtaa aagctctgat
     3361 gtatctatct tttttacacc gttttcatct gtgcatatgg acagttttcc ctttgatatc
     3421 taacggtgaa cagttgttct acttttgttt gttagtcttg atgcttcact gatagataca
     3481 agagccataa gaacctcaga tccttccgta tttagccagt atgttctcta gtgtggttcg
     3541 ttgtttttgc gtgagccatg agaacgaacc attgagatca tgcttacttt gcatgtcact
     3601 caaaaatttt gcctcaaaac tggtgagctg aatttttgca gttaaagcat cgtgtagtgt
     3661 ttttcttagt ccgttacgta ggtaggaatc tgatgtaatg gttgttggta ttttgtcacc
     3721 attcattttt atctggttgt tctcaagttc ggttacgaga tccatttgtc tatctagttc
     3781 aacttggaaa atcaacgtat cagtcgggcg gcctcgctta tcaaccacca atttcatatt
     3841 gctgtaagtg tttaaatctt tacttattgg tttcaaaacc cattggttaa gccttttaaa
     3901 ctcatggtag ttattttcaa gcattaacat gaacttaaat tcatcaaggc taatctctat
     3961 atttgccttg tgagttttct tttgtgttag ttcttttaat aaccactcat aaatcctcat
     4021 agagtatttg ttttcaaaag acttaacatg ttccagatta tattttatga atttttttaa
     4081 ctggaaaaga taaggcaata tctcttcact aaaaactaat tctaattttt cgcttgagaa
     4141 cttggcatag tttgtccact ggaaaatctc aaagccttta accaaaggat tcctgatttc
     4201 cacagttctc gtcatcagct ctctggttgc tttagctaat acaccataag cattttccct
     4261 actgatgttc atcatctgag cgtattggtt ataagtgaac gataccgtcc gttctttcct
     4321 tgtagggttt tcaatcgtgg ggttgagtag tgccacacag cataaaatta gcttggtttc
     4381 atgctccgtt aagtcatagc gactaatcgc tagttcattt gctttgaaaa caactaattc
     4441 agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga gatgggctag
     4501 tcaatgataa ttactagtcc ttttcctttg agttgtgggt atctgtaaat tctgctagac
     4561 ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta gacctttgtg
     4621 tgtttttttt gtttatattc aagtggttat aatttataga ataaagaaag aataaaaaaa
     4681 gataaaaaga atagatccca gccctgtgta taactcacta ctttagtcag ttccgcagta
     4741 ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt aaaaccctaa
     4801 aggcttaagt agcaccctcg caagctcggg caaatcgctg aatattcctt ttgtctccga
     4861 ccatcaggca cctgagtcgc tgtctttttc gtgacattca gttcgctgcg ctcacggctc
     4921 tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt atggattcat gcaaggaaac
     4981 tacccataat acaagaaaag cccgtcacgg gcttctcagg gcgttttatg gcgggtctgc
     5041 tatgtggtgc tatctgactt tttgctgttc agcagttcct gccctctgat tttccagtct
     5101 gaccacttcg gattatcccg tgacaggtca ttcagactgg ctaatgcacc cagtaaggca
     5161 gcggtatcat caacaggctt acccgtctta ctgtccctag tgcttggatt ctcaccaata
     5221 aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt
     5281 actggatcta tcaacaggag tccaagcgag ctcgtaaact tggtctgaca gttaccaatg
     5341 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5401 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5461 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5521 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5581 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5641 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5701 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5761 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5821 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5881 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5941 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6001 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6061 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6121 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6181 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6241 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
//
