LOCUS       sequence-97758-e        6174 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains PIq driving expression RbsR-T, and PI driving expression of
            TreR-T..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6174)
  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 6174)
  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-T,TreR-T]
FEATURES             Location/Qualifiers
     source          1..6174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1053..1071
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1136..1213
                     /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    2487..2718
                     /gene="attP"
                     /label=lambda attP
                     /bound_moiety="lambda integrase"
                     /note="integrase from phage lambda"
     terminator      2753..2839
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3374..4324)
                     /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      4372..4594
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5086..5180)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(5210..6001,6002..6070))
                     /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     5833..5852
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6071..6162)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcaggacagg gtggcgggga tgatgatttg ttgcggttcg ctgcgcccgg ttacctgcgc
       61 gatcaactgg caagccgcct ggcgtccagc ttcggcgtaa ccgggatcta cggttacgat
      121 ctccggatgg aggaatttca ttaacggcgt attaccgacg ctcgccagtt gcaaggtgtc
      181 gatgcgttgc tcttgcaggt atttacttgc gccaagtgcc agcgtgtcgg ttgcgcacag
      241 taaggcggta gtttcaggcg taatcacttt tgcaacgttc tcatagcctt gcttcatagc
      301 aagccctggc agggcggcaa cgggatgcag tttatgcgct ttgcagaacg ccaggtaggc
      361 ttcgtgacgt cgcttaccgg ttgtcacgtc actgtgcggc acgccgagat aactgatatt
      421 acgatgcccc tggtcataca gccgttgcat caggattttg attgcccctt cgtcgtcata
      481 acagaccgaa gcaaagcctt ttgcgtcacg cgccagcaga accagcgatg actgccagtg
      541 ggctaacatt tcttctgtta tgccagtaaa accgaacagc actacgccgt cgatattacg
      601 ccgtttcagc actcccaaat gttcggcaac taattgcggg gaaaactgac tttccatcat
      661 gattgggtcg taaccttgtt catagaacgc tggcagcatg gtttgaacgg cgagattttc
      721 tgacaacgaa tccagacggg taacaatgat ggcgaccact ttgctctgtt tgcccgccag
      781 ttgttgtgcg gcgcggttgg gaatgtaatt gagctccgcc atcgccgctt ccactttttc
      841 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg ttggaaacgg tcgccgtaga
      901 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
      961 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcg ctagcctaag
     1021 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     1081 ttcacctcga caattcgcgc gcgaaggcga agcggcatgc atttacgttg gtaccgacac
     1141 catcgaatgg tgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt
     1201 cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc
     1261 tacggcgacc gtttccaacg tggtgaacca ggccagccac gtttctgcga aaacgcggga
     1321 aaaagtggaa gcggcgatgg cggagctcaa ttacattccc aaccgcgtgg cacaacaact
     1381 ggcgggcaaa gcgagccata ccattggcat gttgatcact gccagtacca atcctttcta
     1441 ttcagaactg gtgcgtggcg ttgaacgcag ctgcttcgaa cgcggttata gtctcgtcct
     1501 ttgcaatacc gaaggcgatg aacagcggat gaatcgcaat ctggaaacgc tgatgcaaaa
     1561 acgcgttgat ggcttgctgt tactgtgcac cgaaacgcat caaccttcgc gtgaaatcat
     1621 gcaacgttat ccgacagtgc ctactgtgat gatggactgg gctccgttcg atggcgacag
     1681 cgatcttatt caggataact cgttgctggg cggagactta gcaacgcaat atctgatcga
     1741 taaaggtcat acccgtatcg cctgtattac cggcccgctg gataaaactc cggcgcgcct
     1801 gcggttggaa ggttatcggg cggcgatgaa acgtgcgggt ctcaacattc ctgatggcta
     1861 tgaagtcact ggtgattttg aatttaacgg cgggtttgac gctatgcgcc aactgctatc
     1921 acatccgctg cgtcctcagg ccgtctttac cggaaatgac gctatggctg ttggcgttta
     1981 ccaggcgtta tatcaggcag agttacaggt tccgcaggat atcgcggtga ttggctatga
     2041 cgatatcgaa ctggcaagct ttatgacgcc accattaacc actatccacc aaccgaaaga
     2101 tgaactgggg gagctggcga ttgatgtact catccatcgg ataacccagc cgacccttca
     2161 gcaacaacga ttacaactta ctccgattct gatggaacgc ggttcggctt agccatgggc
     2221 gcaacgcaat taatgtaagt tagcgcgaat tgtcgaggga aatcataaaa aatttatttg
     2281 ctttcaggaa aatttttctg tataatagat tcataaattt gagagaagag tttaaatatg
     2341 gctggttctc gcagaaagaa acatatccat gaaatcccgc cccgaattca tatgtctaga
     2401 tagctgagta gcctagagtg cgcgacaggt ttgatgacaa aaaattagcg caagaagaca
     2461 aaaatcacct tgcgctaatg ctctgttaca ggtcactaat accatctaag tagttgattc
     2521 atagtgactg catatgttgt gttttacagt attatgtagt ctgtttttta tgcaaaatct
     2581 aatttaatat attgatattt atatcatttt acgtttctcg ttcagctttt ttatactaag
     2641 ttggcattat aaaaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
     2701 aaaataaaat cattatttga tttcaatttt gtcccactcc ccctagaggc atcaaataaa
     2761 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     2821 tctcctgagt aggacaaatc cgccgcccta gacctagggt acgggttttg ctgcccgcaa
     2881 acgggctgtt ctggtgttgc tagtttgtta tcagaatcgc agatccggct tcaggtttgc
     2941 cggctgaaag cgctatttct tccagaattg ccatgatttt ttccccacgg gaggcgtcac
     3001 tggctcccgt gttgtcggca gctttgattc gataagcagc atcgcctgtt tcaggctgtc
     3061 tatgtgtgac tgttgagctg taacaagttg tctcaggtgt tcaatttcat gttctagttg
     3121 ctttgtttta ctggtttcac ctgttctatt aggtgttaca tgctgttcat ctgttacatt
     3181 gtcgatctgt tcatggtgaa cagctttaaa tgcaccaaaa actcgtaaaa gctctgatgt
     3241 atctatcttt tttacaccgt tttcatctgt gcatatggac agttttccct ttgatatcta
     3301 acggtgaaca gttgttctac ttttgtttgt tagtcttgat gcttcactga tagatacaag
     3361 agccataaga acctcagatc cttccgtatt tagccagtat gttctctagt gtggttcgtt
     3421 gtttttgcgt gagccatgag aacgaaccat tgagatcatg cttactttgc atgtcactca
     3481 aaaattttgc ctcaaaactg gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt
     3541 ttcttagtcc gttacgtagg taggaatctg atgtaatggt tgttggtatt ttgtcaccat
     3601 tcatttttat ctggttgttc tcaagttcgg ttacgagatc catttgtcta tctagttcaa
     3661 cttggaaaat caacgtatca gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc
     3721 tgtaagtgtt taaatcttta cttattggtt tcaaaaccca ttggttaagc cttttaaact
     3781 catggtagtt attttcaagc attaacatga acttaaattc atcaaggcta atctctatat
     3841 ttgccttgtg agttttcttt tgtgttagtt cttttaataa ccactcataa atcctcatag
     3901 agtatttgtt ttcaaaagac ttaacatgtt ccagattata ttttatgaat ttttttaact
     3961 ggaaaagata aggcaatatc tcttcactaa aaactaattc taatttttcg cttgagaact
     4021 tggcatagtt tgtccactgg aaaatctcaa agcctttaac caaaggattc ctgatttcca
     4081 cagttctcgt catcagctct ctggttgctt tagctaatac accataagca ttttccctac
     4141 tgatgttcat catctgagcg tattggttat aagtgaacga taccgtccgt tctttccttg
     4201 tagggttttc aatcgtgggg ttgagtagtg ccacacagca taaaattagc ttggtttcat
     4261 gctccgttaa gtcatagcga ctaatcgcta gttcatttgc tttgaaaaca actaattcag
     4321 acatacatct caattggtct aggtgatttt aatcactata ccaattgaga tgggctagtc
     4381 aatgataatt actagtcctt ttcctttgag ttgtgggtat ctgtaaattc tgctagacct
     4441 ttgctggaaa acttgtaaat tctgctagac cctctgtaaa ttccgctaga cctttgtgtg
     4501 ttttttttgt ttatattcaa gtggttataa tttatagaat aaagaaagaa taaaaaaaga
     4561 taaaaagaat agatcccagc cctgtgtata actcactact ttagtcagtt ccgcagtatt
     4621 acaaaaggat gtcgcaaacg ctgtttgctc ctctacaaaa cagaccttaa aaccctaaag
     4681 gcttaagtag caccctcgca agctcgggca aatcgctgaa tattcctttt gtctccgacc
     4741 atcaggcacc tgagtcgctg tctttttcgt gacattcagt tcgctgcgct cacggctctg
     4801 gcagtgaatg ggggtaaatg gcactacagg cgccttttat ggattcatgc aaggaaacta
     4861 cccataatac aagaaaagcc cgtcacgggc ttctcagggc gttttatggc gggtctgcta
     4921 tgtggtgcta tctgactttt tgctgttcag cagttcctgc cctctgattt tccagtctga
     4981 ccacttcgga ttatcccgtg acaggtcatt cagactggct aatgcaccca gtaaggcagc
     5041 ggtatcatca acaggcttac ccgtcttact gtccctagtg cttggattct caccaataaa
     5101 aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac
     5161 tggatctatc aacaggagtc caagcgagct cgtaaacttg gtctgacagt taccaatgct
     5221 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5281 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5341 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5401 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5461 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5521 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5581 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5641 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5701 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5761 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5821 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5881 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5941 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6001 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6061 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6121 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggct cgag
//
