LOCUS       sequence-252138-        2092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the spCas9 Safetargeting Chr11 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 2092)
  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 2092)
  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_058
FEATURES             Location/Qualifiers
     source          1..2092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(84..118)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(189..848)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     763..782
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1030..1618
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1519..1538
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(1927..1960)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1964..1984
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
ORIGIN
        1 agcaagttta aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
       61 ttttttgctg tgagaccaga ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca
      121 aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg
      181 atatcaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
      241 ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc
      301 accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc
      361 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgaaacgaaa
      421 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
      481 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
      541 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
      601 accagctcac cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca
      661 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
      721 gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc
      781 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
      841 ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt
      901 agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgcccgat caatcatgac
      961 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1021 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1081 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1141 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     1201 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1261 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1321 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1381 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1441 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1501 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1561 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     1621 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     1681 ctttcctgcg ttatcccctg attctgtgga taaccgtagt cggtctcaaa cggatccgac
     1741 gcgccatctc taggcccgcg ccggccccct cgcacggact tgtgggagaa gctcggctac
     1801 tcccctgccc cggttaattt gcatataata tttcctagta actatagagg cttaatgtgc
     1861 gataaaagac agataatctg ttctttttaa tactagctac attttacatg ataggcttgg
     1921 atttctataa cttcgtatag catacattat acgaagttat aaacagcaca aaaggaaact
     1981 caccctaact gtaaagtaat tgtgtgtttt gagactataa gtatcccttg gagaaccacc
     2041 ttgtttggat gagaactagt cataggttta agagctaagc tggaaacagc at
//
