LOCUS       sequence-248669-        3258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes 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 3258)
  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 3258)
  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_019
FEATURES             Location/Qualifiers
     source          1..3258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             665..1600
                     /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(1623..1657)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1728..2387)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2302..2321
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2569..3157
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3058..3077
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcggtctcat atggacaaga agtacagcat cggcctggcc atcggcacca actctgtggg
       61 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
      121 caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga
      181 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
      241 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
      301 cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc
      361 catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca
      421 cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc
      481 cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga
      541 caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga
      601 ggaaaacccc atcaacgcca gcggaggtgg cggaagcggc ggtgggggaa gcctcgagcc
      661 atctgctgga gacatgagag ctgccaacct ttggccaagc ccgctcatga tcaaacgctc
      721 taagaagaac agcctggcct tgtccctgac ggccgaccag atggtcagtg ccttgttgga
      781 tgctgagccc cccatactct attccgagta tgatcctacc agacccttca gtgaagcttc
      841 gatgatgggc ttactgacca acctggcaga cagggagctg gttcacatga tcaactgggc
      901 gaagagggtg ccaggctttg tggatttgac cctccatgat caggtccacc ttctagaatg
      961 tgcctggcta gagatcctga tgattggtct agtctggcgc tccatggagc acccagtgaa
     1021 gctactgttt gctcctaact tgctcttgga caggaaccag ggaaaatgtg tagagggcat
     1081 ggtggagatc ttcgacatgc tgctggctac atcatctcgg ttccgcatga tgaatctgca
     1141 gggagaggag tttgtgtgcc tcaaatctat tattttgctt aattctggag tgtacacatt
     1201 tctgtccagc accctgaagt ctctggaaga gaaggaccat atccaccgag tcctggacaa
     1261 gatcacagac actttgatcc acctgatggc caaggcaggc ctgaccctgc agcagcagca
     1321 ccagcggctg gcccagctcc tcctcatcct ctcccacatc aggcacatga gtaacaaagg
     1381 catggagcat ctgtacagca tgaagtgcaa gaacgtggtg cccctctatg acctgctgct
     1441 ggaggcggcg gacgcccacc gcctacatgc gcccactagc cgtggagggg catccgtgga
     1501 ggagacagac caaagccact tggccactgc gggctctact tcatcgcatt ccttgcaaaa
     1561 gtattacatc acgggggagg cagagggttt ccctgccaca gctggatcct gagaccagac
     1621 caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg
     1681 tcattactgg atctatcaac aggagtccaa gcgagctcga tatcaaatta cgccccgccc
     1741 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
     1801 caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
     1861 tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
     1921 aaactggtga aactcaccca gggattggct gaaacgaaaa acatattctc aataaaccct
     1981 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
     2041 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
     2101 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
     2161 gccatacgaa attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga
     2221 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg
     2281 gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc
     2341 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
     2401 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
     2461 tgaaagttgg aacctcttac gtgcccgatc aatcatgacc aaaatccctt aacgtgagtt
     2521 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2581 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2641 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2701 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2761 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2821 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2881 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2941 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3001 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3061 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3121 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3181 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3241 ttctgtggat aaccgtag
//
