LOCUS       sequence-236622-        4920 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes dsaCAS9 as a Type 3b part to be used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4920)
  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 4920)
  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: MTK3b_019
FEATURES             Location/Qualifiers
     source          1..4920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             117..137
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3318..3365
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      complement(3385..3419)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(3490..4149)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4064..4083
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      4331..4919
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4820..4839
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
       61 tctttcctgc gttatcccct gattctgtgg ataaccgtag tcggtctcat tctgccccaa
      121 agaagaagcg gaaggtcggt atccacggag tcccagcagc caagcggaac tacatcctgg
      181 gcctggccat cggcatcacc agcgtgggct acggcatcat cgactacgag acacgggacg
      241 tgatcgatgc cggcgtgcgg ctgttcaaag aggccaacgt ggaaaacaac gagggcaggc
      301 ggagcaagag aggcgccaga aggctgaagc ggcggaggcg gcatagaatc cagagagtga
      361 agaagctgct gttcgactac aacctgctga ccgaccacag cgagctgagc ggcatcaacc
      421 cctacgaggc cagagtgaag ggcctgagcc agaagctgag cgaggaagag ttctctgccg
      481 ccctgctgca cctggccaag agaagaggcg tgcacaacgt gaacgaggtg gaagaggaca
      541 ccggcaacga gctgtccacc aaagagcaga tcagccggaa cagcaaggcc ctggaagaga
      601 aatacgtggc cgaactgcag ctggaacggc tgaagaaaga cggcgaagtg cggggcagca
      661 tcaacagatt caagaccagc gactacgtga aagaagccaa acagctgctg aaggtgcaga
      721 aggcctacca ccagctggac cagagcttca tcgacaccta catcgacctg ctggaaaccc
      781 ggcggaccta ctatgaggga cctggcgagg gcagcccctt cggctggaag gacatcaaag
      841 aatggtacga gatgctgatg ggccactgca cctacttccc cgaggaactg cggagcgtga
      901 agtacgccta caacgccgac ctgtacaacg ccctgaacga cctgaacaat ctcgtgatca
      961 ccagggacga gaacgagaag ctggaatatt acgagaagtt ccagatcatc gagaacgtgt
     1021 tcaagcagaa gaagaagccc accctgaagc agatcgccaa agaaatcctc gtgaacgaag
     1081 aggatattaa gggctacaga gtgaccagca ccggcaagcc cgagttcacc aacctgaagg
     1141 tgtaccacga catcaaggac attaccgccc ggaaagagat tattgagaac gccgagctgc
     1201 tggatcagat tgccaagatc ctgaccatct accagagcag cgaggacatc caggaagaac
     1261 tgaccaatct gaactccgag ctgacccagg aagagatcga gcagatctct aatctgaagg
     1321 gctataccgg cacccacaac ctgagcctga aggccatcaa cctgatcctg gacgagctgt
     1381 ggcacaccaa cgacaaccag atcgctatct tcaaccggct gaagctggtg cccaagaagg
     1441 tggacctgtc ccagcagaaa gagatcccca ccaccctggt ggacgacttc atcctgagcc
     1501 ccgtcgtgaa gagaagcttc atccagagca tcaaagtgat caacgccatc atcaagaagt
     1561 acggcctgcc caacgacatc attatcgagc tggcccgcga gaagaactcc aaggacgccc
     1621 agaaaatgat caacgagatg cagaagcgga accggcagac caacgagcgg atcgaggaaa
     1681 tcatccggac caccggcaaa gagaacgcca agtacctgat cgagaagatc aagctgcacg
     1741 acatgcagga aggcaagtgc ctgtacagcc tggaagccat ccctctggaa gatctgctga
     1801 acaacccctt caactatgag gtggaccaca tcatccccag aagcgtgtcc ttcgacaaca
     1861 gcttcaacaa caaggtgctc gtgaagcagg aagaagccag caagaagggc aaccggaccc
     1921 cattccagta cctgagcagc agcgacagca agatcagcta cgaaaccttc aagaagcaca
     1981 tcctgaatct ggccaagggc aagggcagaa tcagcaagac caagaaagag tatctgctgg
     2041 aagaacggga catcaacagg ttctccgtgc agaaagactt catcaaccgg aacctggtgg
     2101 ataccagata cgccaccaga ggcctgatga acctgctgcg gagctacttc agagtgaaca
     2161 acctggacgt gaaagtgaag tccatcaatg gcggcttcac cagctttctg cggcggaagt
     2221 ggaagtttaa gaaagagcgg aacaaggggt acaagcacca cgccgaggac gccctgatca
     2281 ttgccaacgc cgatttcatc ttcaaagagt ggaagaaact ggacaaggcc aaaaaagtga
     2341 tggaaaacca gatgttcgag gaaaagcagg ccgagagcat gcccgagatc gaaaccgagc
     2401 aggagtacaa agagatcttc atcacccccc accagatcaa gcacattaag gacttcaagg
     2461 actacaagta cagccaccgg gtggacaaga agcctaatag agagctgatt aacgacaccc
     2521 tgtactccac ccggaaggac gacaagggca acaccctgat cgtgaacaat ctgaacggcc
     2581 tgtacgacaa ggacaatgac aagctgaaaa agctgatcaa caagagcccc gaaaagctgc
     2641 tgatgtacca ccacgacccc cagacctacc agaaactgaa gctgattatg gaacagtacg
     2701 gcgacgagaa gaatcccctg tacaagtact acgaggaaac cgggaactac ctgaccaagt
     2761 actccaaaaa ggacaacggc cccgtgatca agaagattaa gtattacggc aacaaactga
     2821 acgcccatct ggacatcacc gacgactacc ccaacagcag aaacaaggtc gtgaagctgt
     2881 ccctgaagcc ctacagattc gacgtgtacc tggacaatgg cgtgtacaag ttcgtgaccg
     2941 tgaagaatct ggatgtgatc aaaaaagaaa actactacga agtgaatagc aagtgctatg
     3001 aggaagctaa gaagctgaag aagatcagca accaggccga gtttatcgcc tccttctaca
     3061 acaacgatct gatcaagatc aacggcgagc tgtatagagt gatcggcgtg aacaacgacc
     3121 tgctgaaccg gatcgaagtg aacatgatcg acatcaccta ccgcgagtac ctggaaaaca
     3181 tgaacgacaa gaggcccccc aggatcatta agacaatcgc ctccaagacc cagagcatta
     3241 agaagtacag cacagacatt ctgggcaacc tgtatgaagt gaaatctaag aagcaccctc
     3301 agatcatcaa aaagggcaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga
     3361 aaaagggatc ctgagaccag accaataaaa aacgcccggc ggcaaccgag cgttctgaac
     3421 aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc
     3481 gatatcaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat
     3541 tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag
     3601 caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc
     3661 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgaaacgaa
     3721 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac
     3781 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
     3841 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
     3901 caccagctca ccgtctttca ttgccatacg aaattccgga tgagcattca tcaggcgggc
     3961 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa
     4021 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc
     4081 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
     4141 tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     4201 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgcccga tcaatcatga
     4261 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4321 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4381 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4441 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4501 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4561 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4621 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4681 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4741 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4801 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4861 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
//
