LOCUS       sequence-249736-        5862 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes NLS::dCAS9::NLS 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 5862)
  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 5862)
  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_030
FEATURES             Location/Qualifiers
     source          1..5862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      80..668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     569..588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             789..809
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4926..4946
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4956..4976
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      complement(4996..5030)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(5101..5760)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5675..5694
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 caatcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
       61 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
      121 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
      181 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      241 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      301 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      361 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
      421 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      481 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      541 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      601 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      661 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      721 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtagt cggtctcatt
      781 ctgacgtccc aaagaagaag agaaaggtag gcagcgacaa gaagtattct atcggactgg
      841 ccatcgggac taatagcgtc gggtgggccg tgatcactga cgagtacaag gtgccctcta
      901 agaagttcaa ggtgctcggg aacaccgacc ggcattccat caagaaaaat ctgatcggag
      961 ctctcctctt tgattcaggg gaaactgctg aagcaacccg cctcaagcgg actgctagac
     1021 ggcggtacac caggaggaag aaccggattt gttaccttca agagatattc tccaacgaaa
     1081 tggcaaaggt cgacgacagc ttcttccata ggctggaaga atcattcctc gtggaagagg
     1141 ataagaagca tgaacggcat cccatcttcg gtaatatcgt cgacgaggtg gcctatcacg
     1201 agaaataccc aaccatctac catcttcgca aaaagctggt ggactcaacc gacaaggcag
     1261 acctccggct tatctacctg gccctggccc acatgatcaa gttcagaggc cacttcctga
     1321 tcgagggcga cctcaatcct gacaatagcg atgtggataa actgttcatc cagctggtgc
     1381 agacttacaa ccagctcttt gaagagaacc ccatcaatgc aagcggagtc gatgccaagg
     1441 ccattctgtc agcccggctg tcaaagagcc gcagacttga gaatcttatc gctcagctgc
     1501 cgggtgaaaa gaaaaatgga ctgttcggga acctgattgc tctttcactt gggctgactc
     1561 ccaatttcaa gtctaatttc gacctggcag aggatgccaa gctgcaactg tccaaggaca
     1621 cctatgatga cgatctcgac aacctcctgg cccagatcgg tgaccaatac gccgaccttt
     1681 tccttgctgc taagaatctt tctgacgcca tcctgctgtc tgacattctc cgcgtgaaca
     1741 ctgaaatcac caaggcccct ctttcagctt caatgattaa gcggtatgat gagcaccacc
     1801 aggacctgac cctgcttaag gcactcgtcc ggcagcagct tccggagaag tacaaggaaa
     1861 tcttctttga ccagtcaaag aatggatacg ccggctacat cgacggaggt gcctcccaag
     1921 aggaatttta taagtttatc aaacctatcc ttgagaagat ggacggcacc gaagagctcc
     1981 tcgtgaaact gaatcgggag gatctgctgc ggaagcagcg cactttcgac aatgggagca
     2041 ttccccacca gatccatctt ggggagcttc acgccatcct tcggcgccaa gaggacttct
     2101 acccctttct taaggacaac agggagaaga ttgagaaaat tctcactttc cgcatcccct
     2161 actacgtggg acccctcgcc agaggaaata gccggtttgc ttggatgacc agaaagtcag
     2221 aagaaactat cactccctgg aacttcgaag aggtggtgga caagggagcc agcgctcagt
     2281 cattcatcga acggatgact aacttcgata agaacctccc caatgagaag gtcctgccga
     2341 aacattccct gctctacgag tactttaccg tgtacaacga gctgaccaag gtgaaatatg
     2401 tcaccgaagg gatgaggaag cccgcattcc tgtcaggcga acaaaagaag gcaattgtgg
     2461 accttctgtt caagaccaat agaaaggtga ccgtgaagca gctgaaggag gactatttca
     2521 agaaaattga atgcttcgac tctgtggaga ttagcggggt cgaagatcgg ttcaacgcaa
     2581 gcctgggtac ctaccatgat ctgcttaaga tcatcaagga caaggatttt ctggacaatg
     2641 aggagaaaga ggacatcctt gaggacattg tcctgactct cactctgttc gaggaccggg
     2701 aaatgatcga ggagaggctt aagacctacg cccatctgtt cgacgataaa gtgatgaagc
     2761 aacttaaacg gagaagatat accggatggg gacgccttag ccgcaaactc atcaacggaa
     2821 tccgggacaa acagagcgga aagaccattc ttgatttcct taagagcgac ggattcgcta
     2881 atcgcaactt catgcaactt atccatgatg attccctgac ctttaaggag gacatccaga
     2941 aggcccaagt gtctggacaa ggtgactcac tgcacgagca tatcgcaaat ctggctggtt
     3001 cacccgctat taagaagggt attctccaga ccgtgaaagt cgtggacgag ctggtcaagg
     3061 tgatgggtcg ccataaacca gagaacattg tcatcgagat ggccagggaa aaccagacta
     3121 cccagaaggg acagaagaac agcagggagc ggatgaaaag aattgaggaa gggattaagg
     3181 agctcgggtc acagatcctt aaagagcacc cggtggaaaa cacccagctt cagaatgaga
     3241 agctctatct gtactacctt caaaatggac gcgatatgta tgtggaccaa gagcttgata
     3301 tcaacaggct ctcagactac gacgtggacg ccatcgtccc tcagagcttc ctcaaagacg
     3361 actcaattga caataaggtg ctgactcgct cagacaagaa ccggggaaag tcagataacg
     3421 tgccctcaga ggaagtcgtg aaaaagatga agaactattg gcgccagctt ctgaacgcaa
     3481 agctgatcac tcagcggaag ttcgacaatc tcactaaggc tgagaggggc ggactgagcg
     3541 aactggacaa agcaggattc attaaacggc aacttgtgga gactcggcag attactaaac
     3601 atgtagccca aatccttgac tcacgcatga ataccaagta cgacgaaaac gacaaactta
     3661 tccgcgaggt gaaggtgatt accctgaagt ccaagctggt cagcgatttc agaaaggact
     3721 ttcaattcta caaagtgcgg gagatcaata actatcatca tgctcatgac gcatatctga
     3781 atgccgtggt gggaaccgcc ctgatcaaga agtacccaaa gctggaaagc gagttcgtgt
     3841 acggagacta caaggtctac gacgtgcgca agatgattgc caaatctgag caggagatcg
     3901 gaaaggccac cgcaaagtac ttcttctaca gcaacatcat gaatttcttc aagaccgaaa
     3961 tcacccttgc aaacggtgag atccggaaga ggccgctcat cgagactaat ggggagactg
     4021 gcgaaatcgt gtgggacaag ggcagagatt tcgctaccgt gcgcaaagtg ctttctatgc
     4081 ctcaagtgaa catcgtgaag aaaaccgagg tgcaaaccgg aggcttttct aaggaatcaa
     4141 tcctccccaa gcgcaactcc gacaagctca ttgcaaggaa gaaggattgg gaccctaaga
     4201 agtacggcgg attcgattca ccaactgtgg cttattctgt cctggtcgtg gctaaggtgg
     4261 aaaaaggaaa gtctaagaag ctcaagagcg tgaaggaact gctgggtatc accattatgg
     4321 agcgcagctc cttcgagaag aacccaattg actttctcga agccaaaggt tacaaggaag
     4381 tcaagaagga ccttatcatc aagctcccaa agtatagcct gttcgaactg gagaatgggc
     4441 ggaagcggat gctcgcctcc gctggcgaac ttcagaaggg taatgagctg gctctcccct
     4501 ccaagtacgt gaatttcctc taccttgcaa gccattacga gaagctgaag gggagccccg
     4561 aggacaacga gcaaaagcaa ctgtttgtgg agcagcataa gcattatctg gacgagatca
     4621 ttgagcagat ttccgagttt tctaaacgcg tcattctcgc tgatgccaac ctcgataaag
     4681 tccttagcgc atacaataag cacagagaca aaccaattcg ggagcaggct gagaatatca
     4741 tccacctgtt caccctcacc aatcttggtg cccctgccgc attcaagtac ttcgacacca
     4801 ccatcgaccg gaaacgctat acctccacca aagaagtgct ggacgccacc ctcatccacc
     4861 agagcatcac cggactttac gaaactcgga ttgacctctc acagctcgga ggggatgagg
     4921 gagctcccaa gaaaaagcgc aaggtaggtt ctccaccaaa gaagaagaga aaggttggat
     4981 cctgagacca gaccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
     5041 tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa
     5101 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac
     5161 atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
     5221 gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc
     5281 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgaaacga aaaacatatt
     5341 ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga
     5401 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt
     5461 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc
     5521 accgtctttc attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg
     5581 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat
     5641 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg
     5701 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat
     5761 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     5821 tatttcatta tggtgaaagt tggaacctct tacgtgcccg at
//
