LOCUS       sequence-251646-        2093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the spCas9 pPLMP1 gRNA expression cassette as a type 234 
            part to be used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2093)
  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 2093)
  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: MTK234_011
FEATURES             Location/Qualifiers
     source          1..2093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     415..434
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(823..856)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     860..880
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     terminator      complement(1073..1107)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1178..1837)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1752..1771
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      join(2019..2093,1..514)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
       61 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
      121 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
      181 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
      241 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
      301 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
      361 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
      421 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
      481 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
      541 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
      601 tgtggataac cgtagtcggt ctcaaacgga tccgacgcgc catctctagg cccgcgccgg
      661 ccccctcgca cggacttgtg ggagaagctc ggctactccc ctgccccggt taatttgcat
      721 ataatatttc ctagtaacta tagaggctta atgtgcgata aaagacagat aatctgttct
      781 ttttaatact agctacattt tacatgatag gcttggattt ctataacttc gtatagcata
      841 cattatacga agttataaac agcacaaaag gaaactcacc ctaactgtaa agtaattgtg
      901 tgttttgaga ctataagtat cccttggaga accaccttgt ttgtactgta tgtacataca
      961 gtgtttaaga gctaagctgg aaacagcata gcaagtttaa ataaggctag tccgttatca
     1021 acttgaaaaa gtggcaccga gtcggtgctt tttttgctgt gagaccagac caataaaaaa
     1081 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg
     1141 atctatcaac aggagtccaa gcgagctcga tatcaaatta cgccccgccc tgccactcat
     1201 cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
     1261 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
     1321 tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga
     1381 aactcaccca gggattggct gaaacgaaaa acatattctc aataaaccct ttagggaaat
     1441 aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga
     1501 aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg
     1561 tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa
     1621 attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt
     1681 gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat
     1741 aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata
     1801 tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa
     1861 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     1921 aacctcttac gtgcccgatc aatcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1981 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2041 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg ttt
//
