LOCUS       sequence-249772-        2722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a nuclear localized iRFP713 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 2722)
  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 2722)
  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_013
FEATURES             Location/Qualifiers
     source          1..2722
                     /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"
     terminator      complement(1832..1866)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1937..2596)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2511..2530
                     /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 gctgaaggtt ctgttgctag acaaccagac ttgttgactt
      901 gtgacgacga accaatccac atcccaggtg ctatccaacc acacggtttg ttgttggctt
      961 tggctgctga catgactatc gttgctggtt ctgacaactt gccagaattg actggtttgg
     1021 ctatcggtgc tttgatcggt agatctgctg ctgacgtttt cgactctgaa actcacaaca
     1081 gattgactat cgctttggct gaaccaggtg ctgctgttgg tgctccaatc actgttggtt
     1141 tcactatgag aaaggacgct ggtttcatcg gttcttggca cagacacgac caattgatct
     1201 tcttggaatt ggaaccacca caaagagatg ttgctgaacc acaagctttc ttcagaagaa
     1261 ctaactctgc tatcagaaga ttgcaagctg ctgaaacttt ggaatctgct tgtgctgctg
     1321 ctgctcaaga agttagaaag atcactggtt tcgacagagt tatgatctac agattcgctt
     1381 ctgacttctc tggtgaagtt atcgctgaag acagatgtgc tgaagttgaa tctaagttgg
     1441 gtttgcacta cccagcttct actgttccag ctcaagctag aagattgtac actatcaacc
     1501 cagttagaat catcccagac atcaactaca gaccagttcc agttactcca gacttgaacc
     1561 cagttactgg tagaccaatc gacttgtctt tcgctatctt gagatctgtt tctccagttc
     1621 acttggaatt catgagaaac atcggtatgc acggtactat gtctatctct atcttgagag
     1681 gtgaaagatt gtggggtttg atcgtttgtc accacagaac tccatactac gttgacttgg
     1741 acggtagaca agcttgtgaa ttggttgctc aagttttggc ttggcaaatc ggtgttatgg
     1801 aagaataact cgagtggctg agaccagacc aataaaaaac gcccggcggc aaccgagcgt
     1861 tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag
     1921 cgagctcgat atcaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat
     1981 taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg
     2041 gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga
     2101 agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg
     2161 aaacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac
     2221 acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc
     2281 agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat
     2341 cccatatcac cagctcaccg tctttcattg ccatacgaaa ttccggatga gcattcatca
     2401 ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct
     2461 ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact
     2521 gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag
     2581 tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     2641 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgcccgatca
     2701 atcatgacca aaatccctta ac
//
