LOCUS       sequence-436815-        5653 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Facilitates genomic integration of tetR expression cassette. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5653)
  AUTHORS   Alcantar MA, English MA, Valeri JA, Collins JJ
  TITLE     A high-throughput synthetic biology approach for studying 
            combinatorial chromatin-based transcriptional regulation.
  JOURNAL   Mol Cell. 2024 Jun 20;84(12):2382-2396.e9. doi: 
            10.1016/j.molcel.2024.05.025.
  PUBMED    38906116
REFERENCE   2  (bases 1 to 5653)
  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: 
            "10.1016/j.molcel.2024.05"; journalName: "Mol Cell"; date: 
            "2024-06-20- 20"; volume: "84"; issue: "12"; pages: "2382-2396.e9"
COMMENT     Sequence Label: pReporter_TetR
FEATURES             Location/Qualifiers
     source          1..5653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1169..1185
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1170..1186
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1215..1835
                     /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(1288..1307)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             1845..1865
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      1881..2068
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1936..1960)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             2481..3284
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     CDS             complement(3926..4741)
                     /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(3926..4741)
                     /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     4649..4668
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4733..4752
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4966..5554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5455..5474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcggccgcca atgctaaatt cgagtgaaac acaggaagat cagaaaatcc tcatttcatc
       61 catattaaca ataatttcaa atgtttattt gcattatttg aaactaggca agacaagcaa
      121 cgaaacgttt ttgaaaattt tgagtatttt caataaattt gtagaggact cagatattga
      181 aaaaaagcta cagcaattaa tacttgataa gaagagtatt gagaagggca acggttcatc
      241 atctcatgga tctgcacatg aacaaacacc agagtcaaac gacgttgaaa ttgaggctac
      301 tgcgccaatt gatgacaata cagacgatga taacaaaccg aagttatctg atgtagaaaa
      361 ggattaaaga tgctaagaga tagtgatgat atttcataaa taatgtaatt ctatatatgt
      421 taattacctt ttttgcgagg catatttatg gtgaaggata agttttgacc atcaaagaag
      481 gttaatgtgg ctgtggtttc agggtccata aagcccacat ggataacatt acccctgaat
      541 tcgcatctag actgatgaga cgtggtagag ccacaaacag ccggtacaag caacgatctc
      601 caggaccatc tgaatcatgc gcggatgaca cgaactcacg acggcgatca cagacattaa
      661 cccacagtac agacactgcg acaacgtggc aattcgtcgc aatacaacgc tcaccctaac
      721 atattttcca attaacctca atatttctct gtcacccggc ctctattttc cattttcttc
      781 tttacccgcc acgcgttttt ttctttcaaa tttttttctt ccttcttctt tttcttccac
      841 gtcctcttgc ataaataaat aaaccgtttt gaaaccaaac tcgcctctct ctctcctttt
      901 tgaaatattt ttgggtttgt ttgatccttt ccttcccaat ctctcttgtt taatatatat
      961 tcatttatat cacgctctct ttttatcttc ctttttttcc tctctcttgt attcttcctt
     1021 cccctttcta ctcaaaccaa gaagaaaaag aaaaggtcaa tctttgttaa agaataggat
     1081 cttctactac atcagctttt agatttttca cgcttactgc ttttttcttc ccaagatcga
     1141 aaatttactg aattaacagg gccccccctc gaggtcgacg gtatcgataa gcttgaagca
     1201 agcctcctga aagtatgtct agattagata aaagtaaagt gattaacagc gcattagagc
     1261 tgcttaatga ggtcggaatc gaaggtttaa caacccgtaa actcgcccag aagctaggtg
     1321 tagagcagcc tacattgtat tggcatgtaa aaaataagcg ggctttgctc gacgccttag
     1381 ccattgagat gttagatagg caccatactc acttttgccc tttagaaggg gaaagctggc
     1441 aagatttttt acgtaataac gctaaaagtt ttagatgtgc tttactaagt catcgcgatg
     1501 gagcaaaagt acatttaggt acacggccta cagaaaaaca gtatgaaact ctcgaaaatc
     1561 aattagcctt tttatgccaa caaggttttt cactagagaa tgcattatat gcactcagcg
     1621 ctgtggggca ttttacttta ggttgcgtat tggaagatca agagcatcaa gtcgctaaag
     1681 aagaaaggga aacacctact actgatagta tgccgccatt attacgacaa gctatcgaat
     1741 tatttgatca ccaaggtgca gagccagcct tcttattcgg ccttgaattg atcatatgcg
     1801 gattagaaaa acaacttaaa tgtgaaagtg gatcaggttc tcctccaaaa aaaaagagaa
     1861 aagttggatc ctaactcgag gcgaatttct tatgatttat gatttttatt attaaataag
     1921 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
     1981 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagcatgagg
     2041 tcgctcttat tgaccacacc tctaccggca tgccgagcaa atgcctgcaa atcgctcccc
     2101 atttcgctgg aaatctgctc gtcagtggtg ctcacactga cgaatcatgt acagatcata
     2161 ccgatgactg cctggcgact cacaactaag caagacagcc ggaaccagcg ccggcgaaca
     2221 ccactgcata tatggcatat cacaacagtc cacgtctcaa gcagttacag agatgttacg
     2281 aaccactagt gcactgcagt acacggtttc cttgaaattt ttttgattcg gtaatctccg
     2341 aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata cgcatatgta
     2401 gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga acaaaaacgt
     2461 gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct gctactcatc
     2521 ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca aacttgtgtg
     2581 cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca ttaggtccca
     2641 aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg gagggcacag
     2701 ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgag gacagaaaat
     2761 ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac agaatagcag
     2821 aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt agcggtttga
     2881 agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta gcagaattgt
     2941 catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac attgcgaaga
     3001 gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga agagatgaag
     3061 gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga gatgcattgg
     3121 gtcaacagta tagaaccgtg gatgatgtgg tttctacagg atctgacatt attattgttg
     3181 gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag
     3241 caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact gtattataag
     3301 taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat cagttattac
     3361 ccgagtagag cacttgaatc cactgccccg ggaatctcgg tcgtaatgat ttctataatg
     3421 acgaaaaaaa aaaaattgga aagaaaaagc ttcatggcct ttataaaaag gaactatcca
     3481 atacctcgcc agaaccaagt aacagtattt tacggggcac aaatcaagaa caataagaca
     3541 ggactgtaaa gatggacgca ttgaactcca aagaacaaca agagttccaa aaagtagtgg
     3601 aacaaaagca aatgaaggat ttcatgcgtt tgtactctaa tctggtagaa agatgtttca
     3661 cagactgtgt caatgacttc acaacatcaa agctaaccaa taaggaacaa acatgcatca
     3721 tgaagtgctc agaaaagttc ttgaagcata gcgaacgtgt agggcagcgt ttccaagaac
     3781 aaaacgctgc cttgggacaa ggcttgggcc gataaggtgt actggcgtat atatatctaa
     3841 ttatgtatct ctggtgtagc ccatttttag catgtaaata taaagaccga gcggccgcgt
     3901 gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca
     3961 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     4021 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     4081 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     4141 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt
     4201 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     4261 ccaaaccgtt attcattcgt gattgcgcct gagcgaggcg aaatacgcga tcgctgttaa
     4321 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     4381 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga
     4441 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     4501 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     4561 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     4621 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     4681 catccatgtt ggaatttaat cgcggcctgg agcaagacgt ttcccgttga atatggctca
     4741 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat
     4801 ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt tgttgaataa
     4861 atcgaacttt tgctgagttg aaggatcagt catgaccaaa atcccttaac gtgagttttc
     4921 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4981 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5041 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5101 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5161 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5221 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5281 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5341 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5401 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5461 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5521 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5581 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5641 tgtggataac cgt
//
