LOCUS       sequence-333658-        4178 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ptac single-level gene expression controller (SLC).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4178)
  AUTHORS   Greco FV, Pandi A, Erb TJ, Grierson CS, Gorochowski TE
  TITLE     Harnessing the central dogma for stringent multi-level control of 
            gene expression.
  JOURNAL   Nat Commun. 2021 Mar 19;12(1):1738. doi: 10.1038/s41467-021-21995-7.
  PUBMED    33741937
REFERENCE   2  (bases 1 to 4178)
  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: "Nat 
            Commun."; date: "2021-03-19"; volume: "12(1):1738. doi"; pages: " 
            10.1038/s41467-021-21995-7"
COMMENT     Sequence Label: pPtac
FEATURES             Location/Qualifiers
     source          1..4178
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(719..1534)
                     /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(719..1534)
                     /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     1442..1461
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1526..1545
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1656..2244
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2145..2164
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2363..2368,2369..2384,2385..2391)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2397..2416
                     /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."
     terminator      complement(3305..3332)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3424..3510)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             join(3688..4178,1..592)
                     /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="MKPVTLYDVAEYAGVSYMTVSRVVNQASHVSAKTREKVEAAMVEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIYSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3707..3726)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ggacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct
       61 gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata
      121 tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc
      181 cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt
      241 tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt
      301 tggtgcggat atctcggtag tgggatacga cgataccgaa gatagctcat gttatatccc
      361 gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt
      421 gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgccag tctcactggt
      481 gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
      541 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagatgtggg
      601 gtccgatcta actaaaaagg ccgctctgcg gccttttttc ttttcacttc accgcggaac
      661 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt
      721 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
      781 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
      841 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
      901 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
      961 aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc
     1021 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     1081 cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
     1141 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     1201 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
     1261 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     1321 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     1381 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
     1441 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     1501 tagagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
     1561 aagcagacag ttttattgtt catgaccaaa atcccttaac gtgagttttc gttccactga
     1621 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1681 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1741 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1801 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1861 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1921 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     1981 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2041 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2101 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2161 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2221 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2281 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     2341 cgtattaccg cctttgggca tgttgacaat taatcatcgg ctcgtataat gtgtggaatt
     2401 gtgagcgctc acaattagta gcccggatgc ggatctagga ggaaggatct gatggtgagt
     2461 aaaggagagg agctgtttac aggcgttgta cccatcttgg ttgagttgga tggggacgtt
     2521 aatggtcata agttttcggt tagcggcgag ggcgaaggcg acgctactta cggcaaattg
     2581 acgcttaagt tcatttgtac tacaggaaaa cttccagtcc cctggccaac tctggttaca
     2641 acacttacat acggggtcca atgtttctcg cgttacccag atcacatgaa gcagcacgac
     2701 ttctttaaat ctgcaatgcc agaaggctat gtgcaagagc gcacaatctt cttcaaggat
     2761 gacgggaact ataagacacg cgccgaggtc aagtttgagg gggatacgtt agtgaatcgc
     2821 attgaattga aaggcattga ctttaaagaa gatggcaaca ttttgggaca taaactggag
     2881 tataactata attcgcacaa cgtgtatatc atggctgaca agcagaagaa cgggattaag
     2941 gtaaatttca agattcgtca caacattgag gatggcagcg tgcaattggc tgaccattat
     3001 caacaaaaca cacccattgg cgacggcccg gttttacttc ctgacaatca ctacctgagt
     3061 acacaatcca aattatcgaa ggacccaaat gagaaacgtg atcacatggt ccttttggag
     3121 ttcgtaaccg cggcggggat tactttaggc atggatgagt tgtataagtg aggcggctaa
     3181 acgattccta gaccgacgta cgatacaact agcgctcggt accaaattcc agaaaagagg
     3241 cctcccgaaa ggggggcctt ttttcgtttt ggtccctgag tacagagtct aagggtcggg
     3301 ggccaaaagg ccatccgtca ggatggcctt ctgcttaatt tgatgcctgg cagtttatgg
     3361 cgggcgtcct gcccgccacc ctccgggccg ttgcttcgca acgttcaaat ccgctcccgg
     3421 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     3481 gtctttcgac tgagcctttc gttttataag ctttatcaag gcgaagcttg atgtcccacg
     3541 ggatgaacga gctccagcat acgttaccga gcaaatatct tgactcatgt ctcagattcc
     3601 cagccgcggc gcgccatcga atggtgaaaa acctttcgcg gtatggcatg atagcgcccg
     3661 gaagagagtc aattcatgga ggtgaatatg aaaccagtaa cgttatacga tgtcgcagag
     3721 tatgccggtg tctcttatat gaccgtttcc cgcgtggtga accaggccag ccacgtttct
     3781 gcgaaaacgc gggaaaaagt ggaagcggcg atggtggagc tgaattacat tcccaaccgc
     3841 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
     3901 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
     3961 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
     4021 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
     4081 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
     4141 gaccagacac ccatcaacag tattatttac tcccatga
//
