LOCUS       sequence-249782-        2611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a nuclear localized and destabilized mRuby2 coexpression 
            cassette as a Type 4a part to be used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2611)
  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 2611)
  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: MTK4a_021
FEATURES             Location/Qualifiers
     source          1..2611
                     /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             768..821
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             828..848
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             864..1571
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTHM
                     ALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAVA
                     KFAGLGGGMDELYK"
     terminator      complement(1721..1755)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1826..2485)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2400..2419
                     /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 ctcaatccgg aagcggagag ggcagaggaa
      781 gtctgctaac atgcggtgac gtcgaggaga atcctggacc tggatccccc aagaagaaga
      841 ggaaggtgga ggacgccgga tccgtgtcca aaggagagga gttaatcaag gaaaacatga
      901 gaatgaaagt tgtcatggag ggctccgtta atggtcacca attcaagtgt acaggggaag
      961 gtgaaggtaa tccttacatg ggtacacaaa ctatgagaat taaagtaatt gaaggcggac
     1021 cactaccatt tgcatttgac attctggcaa cgtcattcat gtacggatca cgaactttca
     1081 tcaagtaccc taaaggtata ccagactttt tcaagcaatc ttttccagag ggttttacat
     1141 gggaaagggt tacaagatac gaagatgggg gtgtcgtcac agttatgcaa gatacttcat
     1201 tagaagatgg ctgccttgtc tatcatgtgc aagtaagagg ggtgaatttt ccttctaacg
     1261 gacctgtgat gcagaaaaag accaaaggtt gggaaccaaa tactgaaatg atgtacccag
     1321 ctgatggagg tttgagaggc tacacacaca tggcgcttaa agttgatggt ggaggtcatt
     1381 tgtcttgtag ttttgttacc acttatcgtt ctaaaaagac tgttggcaat atcaaaatgc
     1441 caggaataca tgctgtagac cacagactag aaagactcga agagagcgat aacgaaatgt
     1501 tcgttgtaca gagagagcat gccgtagcca aatttgctgg cttaggcggt ggtatggatg
     1561 aattgtataa gggatcccat ggcttcccgc cggaggtgga ggagcaggat gatggcgcgc
     1621 tgcccatgtc ttgtgcccag gagagcggga tggaccgtca ccctgcagcc tgtgcttctg
     1681 ctaggatcaa tgtgtaactc gagtggctga gaccagacca ataaaaaacg cccggcggca
     1741 accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag
     1801 gagtccaagc gagctcgata tcaaattacg ccccgccctg ccactcatcg cagtactgtt
     1861 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
     1921 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
     1981 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     2041 gattggctga aacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     2101 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
     2161 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
     2221 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacgaaat tccggatgag
     2281 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
     2341 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag
     2401 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
     2461 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact
     2521 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     2581 gcccgatcaa tcatgaccaa aatcccttaa c
//
