LOCUS       sequence-97742-e        6233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains PI driving expression TreR-L, and PI driving expression of 
            LacI-L.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6233)
  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 6233)
  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[TreR-L,LacI-L]
FEATURES             Location/Qualifiers
     source          1..6233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(952..1029)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1073..1091
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1150..1227
                     /gene="lacI"
                     /label=lacI promoter
     protein_bind    2552..2783
                     /gene="attP"
                     /label=lambda attP
                     /bound_moiety="lambda integrase"
                     /note="integrase from phage lambda"
     terminator      2818..2904
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3439..4389)
                     /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      4437..4659
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5151..5245)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(5275..6066,6067..6135))
                     /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     5898..5917
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6136..6227)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctcgagtcag gacagggtgg cggggatgat gatttgttgc ggttcgctgc gcccggttac
       61 ctgcgcgatc aactggcaag ccgcctggcg tccagcttcg gcgtaaccgg gatctacggt
      121 tacgatctcc ggatggagga atttcattaa cggcgtatta ccgacgctcg ccagttgcaa
      181 ggtgtcgatg cgttgctctt gcaggtattt acttgcgcca agtgccagcg tgtcggttgc
      241 gcacagtaag gcggtagttt caggcgtaat cacttttgca acgttctcat agccttgctt
      301 catagcaagc cctggcaggg cggcaacggg atgcagttta tgcgctttgc agaacgccag
      361 gtaggcttcg tgacgtcgct taccggttgt cacgtcactg tgcggcacgc cgagataact
      421 gatattacga tgcccctggt catacagccg ttgcatcagg attttgattg ccccttcgtc
      481 gtcataacag accgaagcaa agccttttgc gtcacgcgcc agcagaacca gcgatgactg
      541 ccagtgggct aacatttctt ctgttatgcc agtaaaaccg aacagcacta cgccgtcgat
      601 attacgccgt ttcagcactc ccaaatgttc ggcaactaat tgcggggaaa actgactttc
      661 catcatgatt gggtcgtaac cttgttcata gaacgctggc agcatggttt gaacggcgag
      721 attttctgac aacgaatcca gacgggtaac aatgatggcg accactttgc tctgtttgcc
      781 cgccagttgt tgtgcggcgc ggttgggaat gtaattgagc tccgccatcg ccgcttccac
      841 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
      901 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
      961 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     1021 gatggtgtcg ctagcctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     1081 tatcacgagg ccctttcgtc ttcacctcga caattcgcgc gcgaaggcga agcggcatgc
     1141 atttacgttg acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg
     1201 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
     1261 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
     1321 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
     1381 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
     1441 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
     1501 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
     1561 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
     1621 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
     1681 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
     1741 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
     1801 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
     1861 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
     1921 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
     1981 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
     2041 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag
     2101 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
     2161 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg
     2221 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctgtgacca
     2281 tgggcgcaac gcaattaatg taagttagcg cgaattgtcg agggaaatca taaaaaattt
     2341 atttgctttc aggaaaattt ttctgtataa tagattcata aatttgagag aagagtttaa
     2401 atatggctgg ttctcgcaga aagaaacata tccatgaaat cccgccccga attcatatgt
     2461 ctagatagct gagtagccta gagtgcgcga caggtttgat gacaaaaaat tagcgcaaga
     2521 agacaaaaat caccttgcgc taatgctctg ttacaggtca ctaataccat ctaagtagtt
     2581 gattcatagt gactgcatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa
     2641 aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag cttttttata
     2701 ctaagttggc attataaaaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     2761 cagtcaaaat aaaatcatta tttgatttca attttgtccc actcccccta gaggcatcaa
     2821 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     2881 aacgctctcc tgagtaggac aaatccgccg ccctagacct agggtacggg ttttgctgcc
     2941 cgcaaacggg ctgttctggt gttgctagtt tgttatcaga atcgcagatc cggcttcagg
     3001 tttgccggct gaaagcgcta tttcttccag aattgccatg attttttccc cacgggaggc
     3061 gtcactggct cccgtgttgt cggcagcttt gattcgataa gcagcatcgc ctgtttcagg
     3121 ctgtctatgt gtgactgttg agctgtaaca agttgtctca ggtgttcaat ttcatgttct
     3181 agttgctttg ttttactggt ttcacctgtt ctattaggtg ttacatgctg ttcatctgtt
     3241 acattgtcga tctgttcatg gtgaacagct ttaaatgcac caaaaactcg taaaagctct
     3301 gatgtatcta tcttttttac accgttttca tctgtgcata tggacagttt tccctttgat
     3361 atctaacggt gaacagttgt tctacttttg tttgttagtc ttgatgcttc actgatagat
     3421 acaagagcca taagaacctc agatccttcc gtatttagcc agtatgttct ctagtgtggt
     3481 tcgttgtttt tgcgtgagcc atgagaacga accattgaga tcatgcttac tttgcatgtc
     3541 actcaaaaat tttgcctcaa aactggtgag ctgaattttt gcagttaaag catcgtgtag
     3601 tgtttttctt agtccgttac gtaggtagga atctgatgta atggttgttg gtattttgtc
     3661 accattcatt tttatctggt tgttctcaag ttcggttacg agatccattt gtctatctag
     3721 ttcaacttgg aaaatcaacg tatcagtcgg gcggcctcgc ttatcaacca ccaatttcat
     3781 attgctgtaa gtgtttaaat ctttacttat tggtttcaaa acccattggt taagcctttt
     3841 aaactcatgg tagttatttt caagcattaa catgaactta aattcatcaa ggctaatctc
     3901 tatatttgcc ttgtgagttt tcttttgtgt tagttctttt aataaccact cataaatcct
     3961 catagagtat ttgttttcaa aagacttaac atgttccaga ttatatttta tgaatttttt
     4021 taactggaaa agataaggca atatctcttc actaaaaact aattctaatt tttcgcttga
     4081 gaacttggca tagtttgtcc actggaaaat ctcaaagcct ttaaccaaag gattcctgat
     4141 ttccacagtt ctcgtcatca gctctctggt tgctttagct aatacaccat aagcattttc
     4201 cctactgatg ttcatcatct gagcgtattg gttataagtg aacgataccg tccgttcttt
     4261 ccttgtaggg ttttcaatcg tggggttgag tagtgccaca cagcataaaa ttagcttggt
     4321 ttcatgctcc gttaagtcat agcgactaat cgctagttca tttgctttga aaacaactaa
     4381 ttcagacata catctcaatt ggtctaggtg attttaatca ctataccaat tgagatgggc
     4441 tagtcaatga taattactag tccttttcct ttgagttgtg ggtatctgta aattctgcta
     4501 gacctttgct ggaaaacttg taaattctgc tagaccctct gtaaattccg ctagaccttt
     4561 gtgtgttttt tttgtttata ttcaagtggt tataatttat agaataaaga aagaataaaa
     4621 aaagataaaa agaatagatc ccagccctgt gtataactca ctactttagt cagttccgca
     4681 gtattacaaa aggatgtcgc aaacgctgtt tgctcctcta caaaacagac cttaaaaccc
     4741 taaaggctta agtagcaccc tcgcaagctc gggcaaatcg ctgaatattc cttttgtctc
     4801 cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat tcagttcgct gcgctcacgg
     4861 ctctggcagt gaatgggggt aaatggcact acaggcgcct tttatggatt catgcaagga
     4921 aactacccat aatacaagaa aagcccgtca cgggcttctc agggcgtttt atggcgggtc
     4981 tgctatgtgg tgctatctga ctttttgctg ttcagcagtt cctgccctct gattttccag
     5041 tctgaccact tcggattatc ccgtgacagg tcattcagac tggctaatgc acccagtaag
     5101 gcagcggtat catcaacagg cttacccgtc ttactgtccc tagtgcttgg attctcacca
     5161 ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
     5221 attactggat ctatcaacag gagtccaagc gagctcgtaa acttggtctg acagttacca
     5281 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5341 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5401 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5461 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5521 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5581 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5641 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5701 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5761 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5821 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5881 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5941 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6001 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6061 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6121 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6181 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag ggg
//
