LOCUS       sequence-250044-        2083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the saCas9 pTRE 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 2083)
  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 2083)
  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_016
FEATURES             Location/Qualifiers
     source          1..2083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      127..715
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     616..635
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(1024..1057)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1061..1081
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     misc_RNA        1164..1239
                     /label=Sa gRNA scaffold
                     /note="guide RNA scaffold for the Staphylococcus aureus 
                     CRISPR/Cas9 system"
     terminator      complement(1264..1298)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1369..2028)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1943..1962
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gatcttattt cattatggtg aaagttggaa cctcttacgt gcccgatcaa tcatgaccaa
       61 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
      121 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
      181 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      241 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
      301 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      361 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      421 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
      481 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
      541 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
      601 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
      661 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
      721 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
      781 tcctgcgtta tcccctgatt ctgtggataa ccgtagtcgg tctcaaacgg atccgacgcg
      841 ccatctctag gcccgcgccg gccccctcgc acggacttgt gggagaagct cggctactcc
      901 cctgccccgg ttaatttgca tataatattt cctagtaact atagaggctt aatgtgcgat
      961 aaaagacaga taatctgttc tttttaatac tagctacatt ttacatgata ggcttggatt
     1021 tctataactt cgtatagcat acattatacg aagttataaa cagcacaaaa ggaaactcac
     1081 cctaactgta aagtaattgt gtgttttgag actataagta tcccttggag aaccaccttg
     1141 ttggttactc cctatcagtg atagttatag tactctggaa acagaatcta ctataacaag
     1201 gcaaaatgcc gtgtttatct cgtcaacttg ttggcgagat ttttttgctg tgagaccaga
     1261 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag
     1321 gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt acgccccgcc
     1381 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     1441 acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     1501 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     1561 aaaactggtg aaactcaccc agggattggc tgaaacgaaa aacatattct caataaaccc
     1621 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     1681 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc
     1741 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     1801 tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     1861 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     1921 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     1981 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     2041 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agt
//
