LOCUS       sequence-249077-        5832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes dspCAS9 tagged with NLS as a Type 3a part to be used in the 
            MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5832)
  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 5832)
  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: MTK3a_025
FEATURES             Location/Qualifiers
     source          1..5832
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             20..40
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4157..4177
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      complement(4197..4231)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(4302..4961)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4876..4895
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      5143..5731
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5632..5651
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcggtctcat atggacgtcc caaagaagaa gagaaaggta ggcagcgaca agaagtattc
       61 tatcggactg gccatcggga ctaatagcgt cgggtgggcc gtgatcactg acgagtacaa
      121 ggtgccctct aagaagttca aggtgctcgg gaacaccgac cggcattcca tcaagaaaaa
      181 tctgatcgga gctctcctct ttgattcagg ggaaactgct gaagcaaccc gcctcaagcg
      241 gactgctaga cggcggtaca ccaggaggaa gaaccggatt tgttaccttc aagagatatt
      301 ctccaacgaa atggcaaagg tcgacgacag cttcttccat aggctggaag aatcattcct
      361 cgtggaagag gataagaagc atgaacggca tcccatcttc ggtaatatcg tcgacgaggt
      421 ggcctatcac gagaaatacc caaccatcta ccatcttcgc aaaaagctgg tggactcaac
      481 cgacaaggca gacctccggc ttatctacct ggccctggcc cacatgatca agttcagagg
      541 ccacttcctg atcgagggcg acctcaatcc tgacaatagc gatgtggata aactgttcat
      601 ccagctggtg cagacttaca accagctctt tgaagagaac cccatcaatg caagcggagt
      661 cgatgccaag gccattctgt cagcccggct gtcaaagagc cgcagacttg agaatcttat
      721 cgctcagctg ccgggtgaaa agaaaaatgg actgttcggg aacctgattg ctctttcact
      781 tgggctgact cccaatttca agtctaattt cgacctggca gaggatgcca agctgcaact
      841 gtccaaggac acctatgatg acgatctcga caacctcctg gcccagatcg gtgaccaata
      901 cgccgacctt ttccttgctg ctaagaatct ttctgacgcc atcctgctgt ctgacattct
      961 ccgcgtgaac actgaaatca ccaaggcccc tctttcagct tcaatgatta agcggtatga
     1021 tgagcaccac caggacctga ccctgcttaa ggcactcgtc cggcagcagc ttccggagaa
     1081 gtacaaggaa atcttctttg accagtcaaa gaatggatac gccggctaca tcgacggagg
     1141 tgcctcccaa gaggaatttt ataagtttat caaacctatc cttgagaaga tggacggcac
     1201 cgaagagctc ctcgtgaaac tgaatcggga ggatctgctg cggaagcagc gcactttcga
     1261 caatgggagc attccccacc agatccatct tggggagctt cacgccatcc ttcggcgcca
     1321 agaggacttc tacccctttc ttaaggacaa cagggagaag attgagaaaa ttctcacttt
     1381 ccgcatcccc tactacgtgg gacccctcgc cagaggaaat agccggtttg cttggatgac
     1441 cagaaagtca gaagaaacta tcactccctg gaacttcgaa gaggtggtgg acaagggagc
     1501 cagcgctcag tcattcatcg aacggatgac taacttcgat aagaacctcc ccaatgagaa
     1561 ggtcctgccg aaacattccc tgctctacga gtactttacc gtgtacaacg agctgaccaa
     1621 ggtgaaatat gtcaccgaag ggatgaggaa gcccgcattc ctgtcaggcg aacaaaagaa
     1681 ggcaattgtg gaccttctgt tcaagaccaa tagaaaggtg accgtgaagc agctgaagga
     1741 ggactatttc aagaaaattg aatgcttcga ctctgtggag attagcgggg tcgaagatcg
     1801 gttcaacgca agcctgggta cctaccatga tctgcttaag atcatcaagg acaaggattt
     1861 tctggacaat gaggagaaag aggacatcct tgaggacatt gtcctgactc tcactctgtt
     1921 cgaggaccgg gaaatgatcg aggagaggct taagacctac gcccatctgt tcgacgataa
     1981 agtgatgaag caacttaaac ggagaagata taccggatgg ggacgcctta gccgcaaact
     2041 catcaacgga atccgggaca aacagagcgg aaagaccatt cttgatttcc ttaagagcga
     2101 cggattcgct aatcgcaact tcatgcaact tatccatgat gattccctga cctttaagga
     2161 ggacatccag aaggcccaag tgtctggaca aggtgactca ctgcacgagc atatcgcaaa
     2221 tctggctggt tcacccgcta ttaagaaggg tattctccag accgtgaaag tcgtggacga
     2281 gctggtcaag gtgatgggtc gccataaacc agagaacatt gtcatcgaga tggccaggga
     2341 aaaccagact acccagaagg gacagaagaa cagcagggag cggatgaaaa gaattgagga
     2401 agggattaag gagctcgggt cacagatcct taaagagcac ccggtggaaa acacccagct
     2461 tcagaatgag aagctctatc tgtactacct tcaaaatgga cgcgatatgt atgtggacca
     2521 agagcttgat atcaacaggc tctcagacta cgacgtggac gccatcgtcc ctcagagctt
     2581 cctcaaagac gactcaattg acaataaggt gctgactcgc tcagacaaga accggggaaa
     2641 gtcagataac gtgccctcag aggaagtcgt gaaaaagatg aagaactatt ggcgccagct
     2701 tctgaacgca aagctgatca ctcagcggaa gttcgacaat ctcactaagg ctgagagggg
     2761 cggactgagc gaactggaca aagcaggatt cattaaacgg caacttgtgg agactcggca
     2821 gattactaaa catgtagccc aaatccttga ctcacgcatg aataccaagt acgacgaaaa
     2881 cgacaaactt atccgcgagg tgaaggtgat taccctgaag tccaagctgg tcagcgattt
     2941 cagaaaggac tttcaattct acaaagtgcg ggagatcaat aactatcatc atgctcatga
     3001 cgcatatctg aatgccgtgg tgggaaccgc cctgatcaag aagtacccaa agctggaaag
     3061 cgagttcgtg tacggagact acaaggtcta cgacgtgcgc aagatgattg ccaaatctga
     3121 gcaggagatc ggaaaggcca ccgcaaagta cttcttctac agcaacatca tgaatttctt
     3181 caagaccgaa atcacccttg caaacggtga gatccggaag aggccgctca tcgagactaa
     3241 tggggagact ggcgaaatcg tgtgggacaa gggcagagat ttcgctaccg tgcgcaaagt
     3301 gctttctatg cctcaagtga acatcgtgaa gaaaaccgag gtgcaaaccg gaggcttttc
     3361 taaggaatca atcctcccca agcgcaactc cgacaagctc attgcaagga agaaggattg
     3421 ggaccctaag aagtacggcg gattcgattc accaactgtg gcttattctg tcctggtcgt
     3481 ggctaaggtg gaaaaaggaa agtctaagaa gctcaagagc gtgaaggaac tgctgggtat
     3541 caccattatg gagcgcagct ccttcgagaa gaacccaatt gactttctcg aagccaaagg
     3601 ttacaaggaa gtcaagaagg accttatcat caagctccca aagtatagcc tgttcgaact
     3661 ggagaatggg cggaagcgga tgctcgcctc cgctggcgaa cttcagaagg gtaatgagct
     3721 ggctctcccc tccaagtacg tgaatttcct ctaccttgca agccattacg agaagctgaa
     3781 ggggagcccc gaggacaacg agcaaaagca actgtttgtg gagcagcata agcattatct
     3841 ggacgagatc attgagcaga tttccgagtt ttctaaacgc gtcattctcg ctgatgccaa
     3901 cctcgataaa gtccttagcg catacaataa gcacagagac aaaccaattc gggagcaggc
     3961 tgagaatatc atccacctgt tcaccctcac caatcttggt gcccctgccg cattcaagta
     4021 cttcgacacc accatcgacc ggaaacgcta tacctccacc aaagaagtgc tggacgccac
     4081 cctcatccac cagagcatca ccggacttta cgaaactcgg attgacctct cacagctcgg
     4141 aggggatgag ggagctccca agaaaaagcg caaggtaggt tcttgagacc agaccaataa
     4201 aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta
     4261 ctggatctat caacaggagt ccaagcgagc tcgatatcaa attacgcccc gccctgccac
     4321 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg
     4381 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     4441 cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
     4501 gtgaaactca cccagggatt ggctgaaacg aaaaacatat tctcaataaa ccctttaggg
     4561 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc
     4621 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa
     4681 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
     4741 cgaaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac
     4801 ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg
     4861 ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg
     4921 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct
     4981 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     5041 ttggaacctc ttacgtgccc gatcaatcat gaccaaaatc ccttaacgtg agttttcgtt
     5101 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5161 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5221 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5281 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5341 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5401 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5461 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5521 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5581 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5641 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5701 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5761 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5821 ggataaccgt ag
//
