LOCUS       sequence-249767-        2488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a nuclear localized 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 2488)
  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 2488)
  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_010
FEATURES             Location/Qualifiers
     source          1..2488
                     /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(1598..1632)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1703..2362)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2277..2296
                     /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 gtaactcgag tggctgagac cagaccaata aaaaacgccc ggcggcaacc
     1621 gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag
     1681 tccaagcgag ctcgatatca aattacgccc cgccctgcca ctcatcgcag tactgttgta
     1741 attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg
     1801 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg
     1861 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     1921 tggctgaaac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     1981 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     2041 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     2101 cactatccca tatcaccagc tcaccgtctt tcattgccat acgaaattcc ggatgagcat
     2161 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     2221 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     2281 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     2341 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa
     2401 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     2461 cgatcaatca tgaccaaaat cccttaac
//
