LOCUS       sequence-249776-        2473 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a nuclear localized tagBFP 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 2473)
  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 2473)
  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_016
FEATURES             Location/Qualifiers
     source          1..2473
                     /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..824
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             825..845
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             861..1559
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     terminator      complement(1583..1617)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1688..2347)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2262..2281
                     /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 aagcggagct actaacttca
      781 gcctgctgaa gcaggctggt gacgtggagg agaaccctgg acctcccaag aagaagagga
      841 aggtggagga cgccggatcc agcgagctga ttaaggagaa catgcacatg aagctgtaca
      901 tggagggcac cgtggacaac catcacttca agtgcacatc cgagggcgaa ggcaagccct
      961 acgagggcac ccagaccatg agaatcaagg tggtcgaggg cggccctctc cccttcgcct
     1021 tcgacatcct ggctactagc ttcctctacg gcagcaagac cttcatcaac cacacccagg
     1081 gcatccccga cttcttcaag cagtccttcc ctgagggctt cacatgggag agagtcacca
     1141 catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc
     1201 tcatctacaa cgtcaagatc agaggggtga acttcacatc caacggccct gtgatgcaga
     1261 agaaaacact cggctgggag gccttcaccg aaacgctgta ccccgctgac ggcggcctgg
     1321 aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag ccatctgatc gcaaacatca
     1381 agaccacata tagatccaag aaacccgcta agaacctcaa gatgcctggc gtctactatg
     1441 tggactacag actggaaaga atcaaggagg ccaacaacga aacctacgtc gagcagcacg
     1501 aggtggcagt ggccagatac tgcgacctcc ctagcaaact ggggcacaag cttaattaac
     1561 tcgagtggct gagaccagac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa
     1621 atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctcga
     1681 tatcaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
     1741 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
     1801 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
     1861 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gaaacgaaaa
     1921 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
     1981 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg
     2041 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
     2101 ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa
     2161 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
     2221 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
     2281 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
     2341 tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     2401 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgcccgatc aatcatgacc
     2461 aaaatccctt aac
//
