LOCUS       sequence-249079-        4920 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes dsaCAS9 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 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: MTK3a_026
FEATURES             Location/Qualifiers
     source          1..4920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      56..644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     545..564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             762..782
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3963..4010
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      complement(4030..4064)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(4135..4794)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4709..4728
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
       61 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
      121 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
      181 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
      241 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
      301 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
      361 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
      421 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
      481 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
      541 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
      601 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
      661 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
      721 cccctgattc tgtggataac cgtagtcggt ctcatatggc cccaaagaag aagcggaagg
      781 tcggtatcca cggagtccca gcagccaagc ggaactacat cctgggcctg gccatcggca
      841 tcaccagcgt gggctacggc atcatcgact acgagacacg ggacgtgatc gatgccggcg
      901 tgcggctgtt caaagaggcc aacgtggaaa acaacgaggg caggcggagc aagagaggcg
      961 ccagaaggct gaagcggcgg aggcggcata gaatccagag agtgaagaag ctgctgttcg
     1021 actacaacct gctgaccgac cacagcgagc tgagcggcat caacccctac gaggccagag
     1081 tgaagggcct gagccagaag ctgagcgagg aagagttctc tgccgccctg ctgcacctgg
     1141 ccaagagaag aggcgtgcac aacgtgaacg aggtggaaga ggacaccggc aacgagctgt
     1201 ccaccaaaga gcagatcagc cggaacagca aggccctgga agagaaatac gtggccgaac
     1261 tgcagctgga acggctgaag aaagacggcg aagtgcgggg cagcatcaac agattcaaga
     1321 ccagcgacta cgtgaaagaa gccaaacagc tgctgaaggt gcagaaggcc taccaccagc
     1381 tggaccagag cttcatcgac acctacatcg acctgctgga aacccggcgg acctactatg
     1441 agggacctgg cgagggcagc cccttcggct ggaaggacat caaagaatgg tacgagatgc
     1501 tgatgggcca ctgcacctac ttccccgagg aactgcggag cgtgaagtac gcctacaacg
     1561 ccgacctgta caacgccctg aacgacctga acaatctcgt gatcaccagg gacgagaacg
     1621 agaagctgga atattacgag aagttccaga tcatcgagaa cgtgttcaag cagaagaaga
     1681 agcccaccct gaagcagatc gccaaagaaa tcctcgtgaa cgaagaggat attaagggct
     1741 acagagtgac cagcaccggc aagcccgagt tcaccaacct gaaggtgtac cacgacatca
     1801 aggacattac cgcccggaaa gagattattg agaacgccga gctgctggat cagattgcca
     1861 agatcctgac catctaccag agcagcgagg acatccagga agaactgacc aatctgaact
     1921 ccgagctgac ccaggaagag atcgagcaga tctctaatct gaagggctat accggcaccc
     1981 acaacctgag cctgaaggcc atcaacctga tcctggacga gctgtggcac accaacgaca
     2041 accagatcgc tatcttcaac cggctgaagc tggtgcccaa gaaggtggac ctgtcccagc
     2101 agaaagagat ccccaccacc ctggtggacg acttcatcct gagccccgtc gtgaagagaa
     2161 gcttcatcca gagcatcaaa gtgatcaacg ccatcatcaa gaagtacggc ctgcccaacg
     2221 acatcattat cgagctggcc cgcgagaaga actccaagga cgcccagaaa atgatcaacg
     2281 agatgcagaa gcggaaccgg cagaccaacg agcggatcga ggaaatcatc cggaccaccg
     2341 gcaaagagaa cgccaagtac ctgatcgaga agatcaagct gcacgacatg caggaaggca
     2401 agtgcctgta cagcctggaa gccatccctc tggaagatct gctgaacaac cccttcaact
     2461 atgaggtgga ccacatcatc cccagaagcg tgtccttcga caacagcttc aacaacaagg
     2521 tgctcgtgaa gcaggaagaa gccagcaaga agggcaaccg gaccccattc cagtacctga
     2581 gcagcagcga cagcaagatc agctacgaaa ccttcaagaa gcacatcctg aatctggcca
     2641 agggcaaggg cagaatcagc aagaccaaga aagagtatct gctggaagaa cgggacatca
     2701 acaggttctc cgtgcagaaa gacttcatca accggaacct ggtggatacc agatacgcca
     2761 ccagaggcct gatgaacctg ctgcggagct acttcagagt gaacaacctg gacgtgaaag
     2821 tgaagtccat caatggcggc ttcaccagct ttctgcggcg gaagtggaag tttaagaaag
     2881 agcggaacaa ggggtacaag caccacgccg aggacgccct gatcattgcc aacgccgatt
     2941 tcatcttcaa agagtggaag aaactggaca aggccaaaaa agtgatggaa aaccagatgt
     3001 tcgaggaaaa gcaggccgag agcatgcccg agatcgaaac cgagcaggag tacaaagaga
     3061 tcttcatcac cccccaccag atcaagcaca ttaaggactt caaggactac aagtacagcc
     3121 accgggtgga caagaagcct aatagagagc tgattaacga caccctgtac tccacccgga
     3181 aggacgacaa gggcaacacc ctgatcgtga acaatctgaa cggcctgtac gacaaggaca
     3241 atgacaagct gaaaaagctg atcaacaaga gccccgaaaa gctgctgatg taccaccacg
     3301 acccccagac ctaccagaaa ctgaagctga ttatggaaca gtacggcgac gagaagaatc
     3361 ccctgtacaa gtactacgag gaaaccggga actacctgac caagtactcc aaaaaggaca
     3421 acggccccgt gatcaagaag attaagtatt acggcaacaa actgaacgcc catctggaca
     3481 tcaccgacga ctaccccaac agcagaaaca aggtcgtgaa gctgtccctg aagccctaca
     3541 gattcgacgt gtacctggac aatggcgtgt acaagttcgt gaccgtgaag aatctggatg
     3601 tgatcaaaaa agaaaactac tacgaagtga atagcaagtg ctatgaggaa gctaagaagc
     3661 tgaagaagat cagcaaccag gccgagttta tcgcctcctt ctacaacaac gatctgatca
     3721 agatcaacgg cgagctgtat agagtgatcg gcgtgaacaa cgacctgctg aaccggatcg
     3781 aagtgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc
     3841 cccccaggat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag
     3901 acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg
     3961 gcaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag ggttcttgag
     4021 accagaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt
     4081 tctgaggtca ttactggatc tatcaacagg agtccaagcg agctcgatat caaattacgc
     4141 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
     4201 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
     4261 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
     4321 taaatcaaaa ctggtgaaac tcacccaggg attggctgaa acgaaaaaca tattctcaat
     4381 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
     4441 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
     4501 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
     4561 tttcattgcc atacgaaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
     4621 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
     4681 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
     4741 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
     4801 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
     4861 attatggtga aagttggaac ctcttacgtg cccgatcaat catgaccaaa atcccttaac
//
