LOCUS       sequence-248656-        2559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes nuclear localized bacterial EL222 fused to activating domain
            of VP16 as Type 3 part to be used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2559)
  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 2559)
  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: MTK3_008
FEATURES             Location/Qualifiers
     source          1..2559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..600
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     501..520
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             718..738
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             739..972
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     terminator      complement(1625..1659)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1730..2389)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2304..2323
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
       61 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      121 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
      181 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      241 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      301 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      361 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      421 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      481 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      541 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      601 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      661 ttctttcctg cgttatcccc tgattctgtg gataaccgta gtcggtctca tatgggccct
      721 aaaaagaagc gtaaagtcgc ccccccgacc gatgtcagcc tgggggacga gctccactta
      781 gacggcgagg acgtggcgat ggcgcatgcc gacgcgctag acgatttcga tctggacatg
      841 ttgggggacg gggattcccc gggtccggga tttacccccc acgactccgc cccctacggc
      901 gctctggata tggccgactt cgagtttgag cagatgttta ccgatgccct tggaattgac
      961 gagtacggtg gggaattcgg ggcagacgac acacgcgttg aggtgcaacc gccggcgcag
     1021 tgggtcctcg acctgatcga ggccagcccg atcgcatcgg tcgtgtccga tccgcgtcta
     1081 gccgacaatc cgctgatcgc catcaaccag gccttcaccg acctgaccgg ctattccgaa
     1141 gaagaatgcg tcggccgcaa ttgccgattc ctggcaggtt ccggcaccga gccgtggctg
     1201 accgacaaga tccgccaagg cgtgcgcgag cacaagccgg tgctggtcga gatcctgaac
     1261 tacaagaagg acggcacgcc gttccgcaat gccgtgctcg ttgcaccgat ctacgatgac
     1321 gacgacgagc ttctctattt cctcggcagc caggtcgaag tcgacgacga ccagcccaac
     1381 atgggcatgg cgcgccgcga acgcgccgcg gaaatgctca agacgctgtc gccgcgccag
     1441 ctcgaggtta cgacgctggt ggcatcgggc ttgcgcaaca aggaagtggc ggcccggctc
     1501 ggcctgtcgg agaaaaccgt caagatgcac cgcgggctgg tgatggaaaa gctcaacctg
     1561 aagaccagtg ccgatctggt gcgcattgcc gtcgaagccg gaatcggatc ctgagaccag
     1621 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
     1681 ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat tacgccccgc
     1741 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
     1801 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
     1861 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
     1921 caaaactggt gaaactcacc cagggattgg ctgaaacgaa aaacatattc tcaataaacc
     1981 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
     2041 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct
     2101 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
     2161 ttgccatacg aaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
     2221 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
     2281 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
     2341 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
     2401 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
     2461 ggtgaaagtt ggaacctctt acgtgcccga tcaatcatga ccaaaatccc ttaacgtgag
     2521 ttttcgttcc actgagcgtc agaccccgta gaaaagatc
//
