LOCUS       sequence-251859-        2092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the spCas9 Safetargeting Chr20 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_057
FEATURES             Location/Qualifiers
     source          1..2092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      34..622
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     523..542
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(931..964)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     968..988
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     terminator      complement(1180..1214)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1285..1944)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1859..1878
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
       61 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
      121 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
      181 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
      241 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
      301 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
      361 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
      421 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
      481 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
      541 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
      601 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
      661 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
      721 tagtcggtct caaacggatc cgacgcgcca tctctaggcc cgcgccggcc ccctcgcacg
      781 gacttgtggg agaagctcgg ctactcccct gccccggtta atttgcatat aatatttcct
      841 agtaactata gaggcttaat gtgcgataaa agacagataa tctgttcttt ttaatactag
      901 ctacatttta catgataggc ttggatttct ataacttcgt atagcataca ttatacgaag
      961 ttataaacag cacaaaagga aactcaccct aactgtaaag taattgtgtg ttttgagact
     1021 ataagtatcc cttggagaac caccttgttt gggcaagcac aatgtactcg tttaagagct
     1081 aagctggaaa cagcatagca agtttaaata aggctagtcc gttatcaact tgaaaaagtg
     1141 gcaccgagtc ggtgcttttt ttgctgtgag accagaccaa taaaaaacgc ccggcggcaa
     1201 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg
     1261 agtccaagcg agctcgatat caaattacgc cccgccctgc cactcatcgc agtactgttg
     1321 taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat
     1381 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg
     1441 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
     1501 attggctgaa acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
     1561 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
     1621 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
     1681 aacactatcc catatcacca gctcaccgtc tttcattgcc atacgaaatt ccggatgagc
     1741 attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt
     1801 tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc
     1861 aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt
     1921 atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc
     1981 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     2041 cccgatcaat catgaccaaa atcccttaac gtgagttttc gttccactga gc
//
