LOCUS       sequence-252146-        2094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the spCas9 sgRNA1 hROSA26 (site 1) 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_012
FEATURES             Location/Qualifiers
     source          1..2094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     416..435
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(824..857)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     861..881
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     terminator      complement(1075..1109)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1180..1839)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1754..1773
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      join(2021..2094,1..515)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
       61 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
      121 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
      181 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
      241 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
      301 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
      361 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
      421 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      481 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      541 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
      601 ctgtggataa ccgtagtcgg tctcaaacgg atccgacgcg ccatctctag gcccgcgccg
      661 gccccctcgc acggacttgt gggagaagct cggctactcc cctgccccgg ttaatttgca
      721 tataatattt cctagtaact atagaggctt aatgtgcgat aaaagacaga taatctgttc
      781 tttttaatac tagctacatt ttacatgata ggcttggatt tctataactt cgtatagcat
      841 acattatacg aagttataaa cagcacaaaa ggaaactcac cctaactgta aagtaattgt
      901 gtgttttgag actataagta tcccttggag aaccaccttg tttgcgttgg tacggtcctc
      961 cccggtttaa gagctaagct ggaaacagca tagcaagttt aaataaggct agtccgttat
     1021 caacttgaaa aagtggcacc gagtcggtgc tttttttgct gtgagaccag accaataaaa
     1081 aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact
     1141 ggatctatca acaggagtcc aagcgagctc gatatcaaat tacgccccgc cctgccactc
     1201 atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc
     1261 atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc
     1321 catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat caaaactggt
     1381 gaaactcacc cagggattgg ctgaaacgaa aaacatattc tcaataaacc ctttagggaa
     1441 ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta gaaactgccg
     1501 gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct catggaaaac
     1561 ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca ttgccatacg
     1621 aaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
     1681 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
     1741 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
     1801 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
     1861 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
     1921 ggaacctctt acgtgcccga tcaatcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1981 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2041 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtt
//
