LOCUS       sequence-248668-        3537 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes rapamycin and tamoxifen activated split-dCAS9 (N-terminal) 
            as a Type 3 part to be used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3537)
  AUTHORS   Fonseca JP, Bonny AR, Kumar GR, Ng AH, Town J, Wu QC, Aslankoohi E, 
            Chen SY, Dods G, Harrigan P, Osimiri LC, Kistler AL, El-Samad H
  TITLE     A Toolkit for Rapid Modular Construction of Biological Circuits in 
            Mammalian Cells.
  JOURNAL   ACS Synth Biol. 2019 Nov 15;8(11):2593-2606. doi: 
            10.1021/acssynbio.9b00322. Epub 2019 Nov 5.
  PUBMED    31686495
REFERENCE   2  (bases 1 to 3537)
  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/acssynbio.9b00322"; journalName: "ACS Synth Biol"; date: 
            "2019-11-15- 15"; volume: "8"; issue: "11"; pages: "2593-2606"
COMMENT     Sequence Label: MTK3_018
FEATURES             Location/Qualifiers
     source          1..3537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      163..751
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     652..671
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1505..1774
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQ"
     CDS             1796..2731
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     terminator      complement(2754..2788)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(2859..3518)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3433..3452
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
       61 ttggaacctc ttacgtgccc gatcaatcat gaccaaaatc ccttaacgtg agttttcgtt
      121 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
      181 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
      241 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
      301 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
      361 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
      421 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
      481 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
      541 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
      601 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
      661 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
      721 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
      781 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
      841 ggataaccgt agtcggtctc atatggacaa gaagtacagc atcggcctgg ccatcggcac
      901 caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa
      961 ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt
     1021 cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac
     1081 cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt
     1141 ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca
     1201 cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc
     1261 caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct
     1321 gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga
     1381 cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa
     1441 ccagctgttc gaggaaaacc ccatcaacgc cagcggaggt ggcggaagcg gcggtggggg
     1501 aagcgagatg tggcacgagg gactcgaaga ggccagcagg ctgtactttg gcgagaggaa
     1561 cgtcaagggc atgttcgaag tgctggagcc cctccatgcg atgatggaaa ggggcccaca
     1621 gaccctgaag gaaaccagct tcaaccaggc ttacggcagg gacctgatgg aggcacagga
     1681 atggtgcagg aagtacatga agagcggcaa cgtgaaagac ctgacccagg cgtgggacct
     1741 ctactaccac gtgttcagga ggatcagcaa gcagggaggt agcctcgagc catctgctgg
     1801 agacatgaga gctgccaacc tttggccaag cccgctcatg atcaaacgct ctaagaagaa
     1861 cagcctggcc ttgtccctga cggccgacca gatggtcagt gccttgttgg atgctgagcc
     1921 ccccatactc tattccgagt atgatcctac cagacccttc agtgaagctt cgatgatggg
     1981 cttactgacc aacctggcag acagggagct ggttcacatg atcaactggg cgaagagggt
     2041 gccaggcttt gtggatttga ccctccatga tcaggtccac cttctagaat gtgcctggct
     2101 agagatcctg atgattggtc ttgtctggcg ctccatggag cacccagtga agctactgtt
     2161 tgctcctaac ttgctcttgg acaggaacca gggaaaatgt gtagagggca tggtggagat
     2221 cttcgacatg ctgctggcta catcatctcg gttccgcatg atgaatctgc agggagagga
     2281 gtttgtgtgc ctcaaatcta ttattttgct taattctgga gtgtacacat ttctgtccag
     2341 caccctgaag tctctggaag agaaggacca tatccaccga gtcctggaca agatcacaga
     2401 cactttgatc cacctgatgg ccaaggcagg cctgaccctg cagcagcagc accagcggct
     2461 ggcccagctc ctcctcatcc tctcccacat caggcacatg agtaacaaag gcatggagca
     2521 tctgtacagc atgaagtgca agaacgtggt gcccctctat gacctgctgc tggaggcggc
     2581 ggacgcccac cgcctacatg cgcccactag ccgtggaggg gcatccgtgg aggagacaga
     2641 ccaaagccac ttggccactg cgggctctac ttcatcgcat tccttgcaaa agtattacat
     2701 cacgggggag gcagagggtt tccctgccac agctggatcc tgagaccaga ccaataaaaa
     2761 acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg
     2821 gatctatcaa caggagtcca agcgagctcg atatcaaatt acgccccgcc ctgccactca
     2881 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca
     2941 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
     3001 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
     3061 aaactcaccc agggattggc tgaaacgaaa aacatattct caataaaccc tttagggaaa
     3121 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
     3181 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
     3241 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacga
     3301 aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
     3361 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
     3421 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
     3481 atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt agctcct
//
