LOCUS       sequence-252153-        2094 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Encodes the spCas9 sgRNA1 hROSA26 (site 2) 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 Apr 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_013
FEATURES             Location/Qualifiers
     source          1..2094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     415..434
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(823..856)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     860..880
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     terminator      complement(1074..1108)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1179..1838)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1753..1772
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      join(2020..2094,1..514)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
       61 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
      121 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
      181 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
      241 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
      301 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
      361 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
      421 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
      481 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
      541 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
      601 tgtggataac cgtagtcggt ctcaaacgga tccgacgcgc catctctagg cccgcgccgg
      661 ccccctcgca cggacttgtg ggagaagctc ggctactccc ctgccccggt taatttgcat
      721 ataatatttc ctagtaacta tagaggctta atgtgcgata aaagacagat aatctgttct
      781 ttttaatact agctacattt tacatgatag gcttggattt ctataacttc gtatagcata
      841 cattatacga agttataaac agcacaaaag gaaactcacc ctaactgtaa agtaattgtg
      901 tgttttgaga ctataagtat cccttggaga accaccttgt ttgggcgatg acgagatcac
      961 gcggtttaag agctaagctg gaaacagcat agcaagttta aataaggcta gtccgttatc
     1021 aacttgaaaa agtggcaccg agtcggtgct ttttttgctg tgagaccaga ccaataaaaa
     1081 acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg
     1141 gatctatcaa caggagtcca agcgagctcg atatcaaatt acgccccgcc ctgccactca
     1201 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca
     1261 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
     1321 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
     1381 aaactcaccc agggattggc tgaaacgaaa aacatattct caataaaccc tttagggaaa
     1441 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
     1501 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
     1561 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacga
     1621 aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
     1681 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
     1741 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
     1801 atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa
     1861 aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg
     1921 gaacctctta cgtgcccgat caatcatgac caaaatccct taacgtgagt tttcgttcca
     1981 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2041 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gttt
//
