LOCUS       sequence-250040-        2093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the spCas9 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 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_010
FEATURES             Location/Qualifiers
     source          1..2093
                     /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"
     terminator      complement(1274..1308)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1379..2038)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1953..1972
                     /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 ttgtacgttc tctatcactg atagtttaag agctaagctg gaaacagcat agcaagttta
     1201 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttgctg
     1261 tgagaccaga ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
     1321 gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt
     1381 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
     1441 ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
     1501 cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca
     1561 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgaaacgaaa aacatattct
     1621 caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat
     1681 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
     1741 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
     1801 cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa
     1861 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
     1921 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
     1981 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt
     2041 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agt
//
