LOCUS       sequence-249088-        1731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes picornavirus peptide T2A element as a Type 3b part to be 
            used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1731)
  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 1731)
  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: MTK3b_003
FEATURES             Location/Qualifiers
     source          1..1731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             22..75
                     /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"
     terminator      complement(95..129)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(200..859)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     774..793
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1041..1629
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1530..1549
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggtctcatt ctggaagcgg agagggcaga ggaagtctgc taacatgcgg tgacgtcgag
       61 gagaatcctg gacctggatc ctgagaccag accaataaaa aacgcccggc ggcaaccgag
      121 cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc
      181 aagcgagctc gatatcaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt
      241 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
      301 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga
      361 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
      421 ctgaaacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
      481 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
      541 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
      601 tatcccatat caccagctca ccgtctttca ttgccatacg aaattccgga tgagcattca
      661 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
      721 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
      781 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
      841 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
      901 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgcccga
      961 tcaatcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1021 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1081 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1141 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     1201 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1261 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     1321 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     1381 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     1441 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     1501 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     1561 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     1621 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     1681 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtag t
//
