LOCUS       sequence-251628-        2094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the spCas9 pC120 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 2094)
  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 2094)
  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_009
FEATURES             Location/Qualifiers
     source          1..2094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      128..716
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     617..636
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(1025..1058)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1062..1082
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     terminator      complement(1276..1310)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1381..2040)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1955..1974
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tgatcttatt tcattatggt gaaagttgga acctcttacg tgcccgatca atcatgacca
       61 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
      121 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
      181 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      241 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
      301 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      361 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      421 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      481 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
      541 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
      601 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
      661 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
      721 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
      781 ttcctgcgtt atcccctgat tctgtggata accgtagtcg gtctcaaacg gatccgacgc
      841 gccatctcta ggcccgcgcc ggccccctcg cacggacttg tgggagaagc tcggctactc
      901 ccctgccccg gttaatttgc atataatatt tcctagtaac tatagaggct taatgtgcga
      961 taaaagacag ataatctgtt ctttttaata ctagctacat tttacatgat aggcttggat
     1021 ttctataact tcgtatagca tacattatac gaagttataa acagcacaaa aggaaactca
     1081 ccctaactgt aaagtaattg tgtgttttga gactataagt atcccttgga gaaccacctt
     1141 gtttggcctt tagtccatgc gttatgttta agagctaagc tggaaacagc atagcaagtt
     1201 taaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttgc
     1261 tgtgagacca gaccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
     1321 tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa
     1381 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac
     1441 atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
     1501 gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc
     1561 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgaaacga aaaacatatt
     1621 ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga
     1681 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt
     1741 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc
     1801 accgtctttc attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg
     1861 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat
     1921 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg
     1981 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat
     2041 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtag
//
