LOCUS       sequence-400056-        6980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SR latch circuit on pLW555 backbone: PTet-A1, PTac-P2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6980)
  AUTHORS   Lebovich M, Zeng M, Andrews LB
  TITLE     Algorithmic Programming of Sequential Logic and Genetic Circuits for
            Recording Biochemical Concentration in a Probiotic Bacterium.
  JOURNAL   ACS Synth Biol. 2023 Aug 15. doi: 10.1021/acssynbio.3c00232.
  PUBMED    37581922
REFERENCE   2  (bases 1 to 6980)
  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"; journalName: "ACS Synth 
            Biol. 2023 Aug 15. doi: 10.1021/acssynbio.3c00232."
COMMENT     Sequence Label: pML15
FEATURES             Location/Qualifiers
     source          1..6980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        89..123
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     RBS             132..143
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             999..1010
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1128..2210
                     /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"
     primer_bind     complement(1147..1166)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2229..2852
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(2302..2321)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3035..3054)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(3419..3964)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3499..3518)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4053..4868)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4053..4868)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4776..4795
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4860..4879
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    5120..5138
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5145..5163
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        join(6095..6100,6101..6116,6117..6123)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    6129..6148
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by 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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
ORIGIN
        1 cgaccaaagg aggtaagtga tgccattgcc tctgaaaggg gtaagacgcc ttctgaattg
       61 tgcagcttaa tggtcaccat atatcaagtt tacggctagc tcagtcctag gtactatgct
      121 agctactaga gaaagaggag aaatactaga tggctgaagc gcaaaatgat cccctgctgc
      181 cgggatactc gtttaatgcc catctggtgg cgggtttaac gccgattgag gccaacggtt
      241 atctcgattt ttttatcgac cgaccgctgg gaatgaaagg ttatattctc aatctcacca
      301 ttcgcggtca gggggtggtg aaaaatcagg gacgagaatt tgtttgccga ccgggtgata
      361 ttttgctgtt cccgccagga gagattcatc actacggtcg tcatccggag gctcgcgaat
      421 ggtatcacca gtgggtttac tttcgtccgc gcgcctactg gcatgaatgg cttaactggc
      481 cgtcaatatt tgccaatacg gggttctttc gcccggatga agcgcaccag ccgcatttca
      541 gcgacctgtt tgggcaaatc attaacgccg ggcaagggga agggcgctat tcggagctgc
      601 tggcgataaa tctgcttgag caattgttac tgcggcgcat ggaagcgatt aacgagtcgc
      661 tccatccacc gatggataat cgggtacgcg aggcttgtca gtacatcagc gatcacctgg
      721 cagacagcaa ttttgatatc gccagcgtcg cacagcatgt ttgcttgtcg ccgtcgcgtc
      781 tgtcacatct tttccgccag cagttaggga ttagcgtctt aagctggcgc gaggaccaac
      841 gtatcagcca ggcgaagctg cttttgagca ccacccggat gcctatcgcc accgtcggtc
      901 gcaatgttgg ttttgacgat caactctatt tctcgcgggt atttaaaaaa tgcaccgggg
      961 ccagcccgag cgagttccgt gccggttaat aaccaattat tgaaggccgc taacgcagcc
     1021 tttttttgtt tctggtctcc caatggcggc gcgccatcga atggcgcaaa acctttcgcg
     1081 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatatg aaaccagtaa
     1141 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     1201 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     1261 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     1321 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     1381 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     1441 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     1501 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
     1561 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgagg
     1621 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
     1681 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc
     1741 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
     1801 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg
     1861 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
     1921 gtgcggatat ctcggtagtg ggatacgacg ataccgaaga tagctcatgt tatatcccgc
     1981 cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     2041 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgccagtc tcactggtga
     2101 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     2161 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga caatccagga
     2221 ggaaaaaaat gtccagatta gataaaagta aagtgattaa cagcgcatta gagctgctta
     2281 atgaggtcgg aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc
     2341 agcctacatt gtattggcat gtaaaaaata agcgggcttt gctcgacgcc ttagccattg
     2401 agatgttaga taggcaccat actcactttt gccctttaga aggggaaagc tggcaagatt
     2461 ttttacgtaa taacgctaaa agttttagat gtgctttact aagtcatcgc gatggagcaa
     2521 aagtacattt aggtacacgg cctacagaaa aacagtatga aactctcgaa aatcaattag
     2581 cctttttatg ccaacaaggt ttttcactag agaatgcatt atatgcactc agcgctgtgg
     2641 ggcattttac tttaggttgc gtattggaag atcaagagca tcaagtcgct aaagaagaaa
     2701 gggaaacacc tactactgat agtatgccgc cattattacg acaagctatc gaattatttg
     2761 atcaccaagg tgcagagcca gccttcttat tcggccttga attgatcata tgcggattag
     2821 aaaaacaact taaatgtgaa agtgggtcct aataattggt aacgaatcag acaattgacg
     2881 gctcgaggga gtagcatagg gtttgcagaa tccctgcttc gtccatttga caggcacatt
     2941 atgcatcgat gataagctgt caaacatgag cagatcctct acgccggacg catcgtggcc
     3001 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg
     3061 gaagatcggg ctcgccactt cgggctcatg agcaaatatt ttatctgagg tgcttcctcg
     3121 ctcactgact cgctgcacga ggcagacctc agcgctagcg gagtgtatac tggcttacta
     3181 tgttggcact gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca
     3241 ccggtgcgtc agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg
     3301 ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc
     3361 ccggaagatg ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt
     3421 tccataggct ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc
     3481 gaaacccgac aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct
     3541 ctcctgttcc tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc
     3601 attccacgcc tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac
     3661 gaaccccccg ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3721 ccggaaagac atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt
     3781 agtcttgaag tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc
     3841 tcctccaagc cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc
     3901 gccctgcaag gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc
     3961 tcaagaagat catcttatta aggggtctga cgctcagtgg aacgaaaaat caatctaaag
     4021 tatatatgag taaacttggt ctgacagtta ccttagaaaa actcatcgag catcaaatga
     4081 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
     4141 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
     4201 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
     4261 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta
     4321 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     4381 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg
     4441 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     4501 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc
     4561 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     4621 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     4681 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     4741 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     4801 aaatcagcat ccatgttgga atttaatcgc ggcctggagc aagacgtttc ccgttgaata
     4861 tggctcataa caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat
     4921 gatatatttt tatcttgtgc aatgtacatc agagattttg agacacaacc aattattgaa
     4981 ggcctcccta acggggggcc tttttttgtt tctggtctcc cgcttggagc gacgtacggt
     5041 ggaatctgat tcgttaccaa ttgacatgat acgaaacgta ccgtatcgtt aaggttacgt
     5101 actccaccgt tggctttttt ccctatcagt gatagagatt gacatcccta tcagtgatag
     5161 agataatgag cacctgaggg tgtctcaagg tgcgtacctt gactgatgag tccgaaagga
     5221 cgaaacaccc ctctacaaat aattttgttt aaaatgttcc ctaataatca gcaaagaggt
     5281 tactagatgg caggcgcagt tggtcgtccg cgtcgtagtg caccgcgtcg tgcaggtaaa
     5341 aatccgcgtg aagaaattct ggatgcaagc gcagaactgt ttacccgtca gggttttgca
     5401 accaccagta cccatcagat tgcagatgca gttggtattc gtcaggcaag cctgtattat
     5461 cattttccga gcaaaaccga aatctttctg accctgctga aaagcaccgt tgaaccgagc
     5521 accgttctgg cagaagatct gagcaccctg gatgcaggtc cggaaatgcg tctgtgggca
     5581 attgttgcaa gcgaagttcg tctgctgctg agcaccaaat ggaatgttgg tcgtctgtat
     5641 cagctgccga ttgttggtag cgaagaattt gcagaatatc atagccagcg tgaagcactg
     5701 accaatgttt ttcgtgatct ggcaaccgaa attgttggtg atgatccgcg tgcagaactg
     5761 ccgtttcata ttaccatgag cgttattgaa atgcgtcgca atgatggtaa aattccgagt
     5821 ccgctgagcg cagatagcct gccggaaacc gcaattatgc tggcagatgc aagcctggca
     5881 gttctgggtg caccgctgcc tgcagatcgt gttgaaaaaa ccctggaact gattaaacag
     5941 gcagatgcaa aataactcgg taccaaagac gaacaataag acgctgaaaa gcgtcttttt
     6001 tcgttttggt ccaggttaag gacgattatg gacagcgggg gggccacgag cctttgagtg
     6061 tctgcgcttc agaggagaac gatcgttggc tgtgttgaca attaatcatc ggctcgtata
     6121 atgtgtggaa ttgtgagcgc tcacaattta cgcttgtcca accaaatgat tcgttaccaa
     6181 ttgacagttt ctatcgatct atagataatg ctagcctgag cggtcaacgc atgtgctttg
     6241 cgttctgatg agacagtgat gtcgaaaccg cctctacaaa taattttgtt taaggagcta
     6301 tggactatgt ttgaaaggct gaaatactag atggcacgta ccccgagccg tagcagcatt
     6361 ggtagcctgc gtagtccgca tacccataaa gcaattctga ccagcaccat tgaaatcctg
     6421 aaagaatgtg gttatagcgg tctgagcatt gaaagcgttg cacgtcgtgc cggtgcaagc
     6481 aaaccgacca tttatcgttg gtggaccaat aaagcagcac tgattgccga agtgtatgaa
     6541 aatgaaagcg aacaggtgcg taaatttccg gatctgggta gctttaaagc cgatctggat
     6601 tttctgctgc gtaatctgtg gaaagtttgg cgtgaaacca tttgtggtga agcatttcgt
     6661 tgtgttattg cagaagcaca gctggaccct gcaaccctga cccagctgaa agatcagttt
     6721 atggaacgtc gtcgtgagat gccgaaaaaa ctggttgaaa atgccattag caatggtgaa
     6781 ctgccgaaag ataccaatcg tgaactgctg ctggatatga tttttggttt ttgttggtat
     6841 cgcctgctga ccgaacagct gaccgttgaa caggatattg aagaatttac cttcctgctg
     6901 attaatggtg tttgtccggg tacacagcgt taaggaaaca cagaaaaaag cccgcacctg
     6961 acagtgcggg cttttttttt
//
