LOCUS       sequence-97741-e        6374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains PIq driving expression GalS-L, and PI driving expression of
            LacI-L..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6374)
  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 6374)
  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,LacI-L]
FEATURES             Location/Qualifiers
     source          1..6374
                     /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
     CDS             1336..2418
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    2693..2924
                     /gene="attP"
                     /label=lambda attP
                     /bound_moiety="lambda integrase"
                     /note="integrase from phage lambda"
     terminator      2959..3045
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3580..4530)
                     /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      4578..4800
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5292..5386)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(5416..6207,6208..6276))
                     /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     6039..6058
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6277..6368)
                     /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 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa
     1501 ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg
     1561 ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg
     1621 gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc
     1681 gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct
     1741 gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc
     1801 atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc
     1861 gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt
     1921 ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa
     1981 cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag
     2041 ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc
     2101 gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat
     2161 accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg
     2221 ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc
     2281 aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa
     2341 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     2401 ctggaaagcg ggcagtgacc atgggcgcaa cgcaattaat gtaagttagc gcgaattgtc
     2461 gagggaaatc ataaaaaatt tatttgcttt caggaaaatt tttctgtata atagattcat
     2521 aaatttgaga gaagagttta aatatggctg gttctcgcag aaagaaacat atccatgaaa
     2581 tcccgccccg aattcatatg tctagatagc tgagtagcct agagtgcgcg acaggtttga
     2641 tgacaaaaaa ttagcgcaag aagacaaaaa tcaccttgcg ctaatgctct gttacaggtc
     2701 actaatacca tctaagtagt tgattcatag tgactgcata tgttgtgttt tacagtatta
     2761 tgtagtctgt tttttatgca aaatctaatt taatatattg atatttatat cattttacgt
     2821 ttctcgttca gcttttttat actaagttgg cattataaaa aagcattgct tatcaatttg
     2881 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgatttc aattttgtcc
     2941 cactccccct agaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     3001 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gccctagacc
     3061 tagggtacgg gttttgctgc ccgcaaacgg gctgttctgg tgttgctagt ttgttatcag
     3121 aatcgcagat ccggcttcag gtttgccggc tgaaagcgct atttcttcca gaattgccat
     3181 gattttttcc ccacgggagg cgtcactggc tcccgtgttg tcggcagctt tgattcgata
     3241 agcagcatcg cctgtttcag gctgtctatg tgtgactgtt gagctgtaac aagttgtctc
     3301 aggtgttcaa tttcatgttc tagttgcttt gttttactgg tttcacctgt tctattaggt
     3361 gttacatgct gttcatctgt tacattgtcg atctgttcat ggtgaacagc tttaaatgca
     3421 ccaaaaactc gtaaaagctc tgatgtatct atctttttta caccgttttc atctgtgcat
     3481 atggacagtt ttccctttga tatctaacgg tgaacagttg ttctactttt gtttgttagt
     3541 cttgatgctt cactgataga tacaagagcc ataagaacct cagatccttc cgtatttagc
     3601 cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg aaccattgag
     3661 atcatgctta ctttgcatgt cactcaaaaa ttttgcctca aaactggtga gctgaatttt
     3721 tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta cgtaggtagg aatctgatgt
     3781 aatggttgtt ggtattttgt caccattcat ttttatctgg ttgttctcaa gttcggttac
     3841 gagatccatt tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg ggcggcctcg
     3901 cttatcaacc accaatttca tattgctgta agtgtttaaa tctttactta ttggtttcaa
     3961 aacccattgg ttaagccttt taaactcatg gtagttattt tcaagcatta acatgaactt
     4021 aaattcatca aggctaatct ctatatttgc cttgtgagtt ttcttttgtg ttagttcttt
     4081 taataaccac tcataaatcc tcatagagta tttgttttca aaagacttaa catgttccag
     4141 attatatttt atgaattttt ttaactggaa aagataaggc aatatctctt cactaaaaac
     4201 taattctaat ttttcgcttg agaacttggc atagtttgtc cactggaaaa tctcaaagcc
     4261 tttaaccaaa ggattcctga tttccacagt tctcgtcatc agctctctgg ttgctttagc
     4321 taatacacca taagcatttt ccctactgat gttcatcatc tgagcgtatt ggttataagt
     4381 gaacgatacc gtccgttctt tccttgtagg gttttcaatc gtggggttga gtagtgccac
     4441 acagcataaa attagcttgg tttcatgctc cgttaagtca tagcgactaa tcgctagttc
     4501 atttgctttg aaaacaacta attcagacat acatctcaat tggtctaggt gattttaatc
     4561 actataccaa ttgagatggg ctagtcaatg ataattacta gtccttttcc tttgagttgt
     4621 gggtatctgt aaattctgct agacctttgc tggaaaactt gtaaattctg ctagaccctc
     4681 tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat attcaagtgg ttataattta
     4741 tagaataaag aaagaataaa aaaagataaa aagaatagat cccagccctg tgtataactc
     4801 actactttag tcagttccgc agtattacaa aaggatgtcg caaacgctgt ttgctcctct
     4861 acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc ctcgcaagct cgggcaaatc
     4921 gctgaatatt ccttttgtct ccgaccatca ggcacctgag tcgctgtctt tttcgtgaca
     4981 ttcagttcgc tgcgctcacg gctctggcag tgaatggggg taaatggcac tacaggcgcc
     5041 ttttatggat tcatgcaagg aaactaccca taatacaaga aaagcccgtc acgggcttct
     5101 cagggcgttt tatggcgggt ctgctatgtg gtgctatctg actttttgct gttcagcagt
     5161 tcctgccctc tgattttcca gtctgaccac ttcggattat cccgtgacag gtcattcaga
     5221 ctggctaatg cacccagtaa ggcagcggta tcatcaacag gcttacccgt cttactgtcc
     5281 ctagtgcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa
     5341 tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcgta
     5401 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5461 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5521 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5581 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5641 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5701 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5761 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5821 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5881 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5941 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6001 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6061 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6121 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6181 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6241 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6301 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6361 taaacaaata gggg
//
