LOCUS       sequence-491937-        3912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes ERK SPARK, a ERK activity reporter, as a Type 3 part to be 
            used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3912)
  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 3912)
  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: MTK3_030
FEATURES             Location/Qualifiers
     source          1..3912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     366..385
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(727..729,730..732,733..1443)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1639..1692
                     /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"
     regulatory      1699..1708
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1909..1911,1912..1914,1915..2625)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      complement(2843..2877)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(2948..3607)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3522..3541
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      join(3789..3912,1..465)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
       61 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
      121 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
      181 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
      241 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
      301 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
      361 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
      421 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
      481 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
      541 tcccctgatt ctgtggataa ccgtagtcgg tctcatatgg ctgaccaact gactgaagag
      601 tggtcgacgg taccgcggac cggtcctgac gtaccacgta caccagtagg aaaggcaaag
      661 ctgtcattcc aattcccggg cggactcgag tccggactca gatctggcag cggtggaggc
      721 gatatcatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      781 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      841 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
      901 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
      961 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     1021 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     1081 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     1141 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     1201 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     1261 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1321 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     1381 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1441 aagggatccg gcagtgctgg tggtagtgct ggtggtagtg ctggtggtag tgctggtggc
     1501 agtgctggtg gtagtgctgg tggtagtgct ggtggctcta gaggagaaat cgcaaagagt
     1561 ctgaaagaaa ttgcaaaatc cttgaaggaa atagcgtggt cactgaaaga aattgccaag
     1621 tccctcaaag ggggtaccga gggcagagga agtctgctaa catgcggtga cgtcgaggag
     1681 aatcctggcc caaagcttgc caccatggcg gacgaggaga agctgccgcc cggctgggag
     1741 aagcgcatga gccgcagctc aggccgagtg tactacttca accacatcac taacgccagc
     1801 cagtgggagc ggcccagcgg caacagcagc agtggtggca aaaacgggca gggggagcct
     1861 gccaggctcg agtccggact cagatctggc agcggtggag gcgatatcat ggtgagcaag
     1921 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1981 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     2041 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2101 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     2161 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2221 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2281 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2341 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggcc
     2401 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     2461 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     2521 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2581 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggatc cggcagtgct
     2641 ggtggtagtg ctggtggtag tgctggtggt agtgctggtg gcagtgctgg tggtagtgct
     2701 ggtggtagtg ctggtggctc tagaaccttg agagaaatcg aggaattgtt gaggaaaatc
     2761 attgaagaca gcgtgcgcag tgtggccgag ctggaggaca ttgagaagtg gctgaagaaa
     2821 atcggatcct gagaccagac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa
     2881 atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctcga
     2941 tatcaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
     3001 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
     3061 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
     3121 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gaaacgaaaa
     3181 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
     3241 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg
     3301 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
     3361 ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa
     3421 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
     3481 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
     3541 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
     3601 tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     3661 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgcccgatc aatcatgacc
     3721 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3781 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3841 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3901 actggcttca gc
//
