LOCUS       sequence-249015-        2382 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes msGFP as a Type 3a part to be used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2382)
  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 2382)
  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: MTK3a_013
FEATURES             Location/Qualifiers
     source          1..2382
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     340..359
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(551..553,554..556,557..1267)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(596..617)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1204..1225
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      complement(1287..1321)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1392..2051)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1966..1985
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      join(2233..2382,1..439)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
       61 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
      121 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
      181 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      241 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      301 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      361 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      421 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
      481 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtag
      541 tcggtctcat atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      601 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg cgcggcgagg gcgagggcga
      661 tgccaccaac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      721 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
      781 ccacatgaag cgccacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      841 caccatcagc ttcaaggacg acggcaccta caagacccgc gccgaggtga agttcgaggg
      901 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
      961 cctggggcac aagctggagt acaacttcaa cagccacaac gtctatatca ccgccgacaa
     1021 gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacgtggagg acggcagcgt
     1081 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1141 cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga
     1201 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actcacggca tggacgagct
     1261 gtacaagggt tcttgagacc agaccaataa aaaacgcccg gcggcaaccg agcgttctga
     1321 acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc
     1381 tcgatatcaa attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc
     1441 attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc
     1501 agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg
     1561 tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgaaacg
     1621 aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc
     1681 acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc
     1741 gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat
     1801 atcaccagct caccgtcttt cattgccata cgaaattccg gatgagcatt catcaggcgg
     1861 gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa
     1921 aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat
     1981 gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt
     2041 tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc
     2101 ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccc gatcaatcat
     2161 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2221 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2281 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2341 ggtaactggc ttcagcagag cgcagatacc aaatactgtt ct
//
