LOCUS       sequence-384576-        2449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Toolkit part 4a encoding a T2A polycistronic element 
            followed by mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2449)
  AUTHORS   Weinberg ZY, Hilburger CE, Kim M, Cao L, Khalid M, Elmes S, Diwanji 
            D, Hernandez E, Lopez J, Schaefer K, Smith AM, Zhou F, QCRG 
            Structural Biology Consortium, Kumar GR, Ott M, Baker D, El-Samad H
  TITLE     Sentinel cells enable genetic detection of SARS-CoV-2 Spike protein
  JOURNAL   bioRxiv 2021.04.20.440678
REFERENCE   2  (bases 1 to 2449)
  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"; journalName: "bioRxiv 
            2021.04.20.440678"
COMMENT     Sequence Label: T2A-mCherry (MTK 4a) (pAN3844)
FEATURES             Location/Qualifiers
     source          1..2449
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      112..700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     601..620
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             824..877
                     /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"
     CDS             887..1597
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1032..1050)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1300..1319
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      complement(1615..1649)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1720..2379)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2294..2313
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tggtgaaagt tggaacctct tacgtgcccg atcaatcatg accaaaatcc cttaacgtga
       61 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
      121 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
      181 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
      241 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
      301 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
      361 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
      421 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
      481 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
      541 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
      601 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
      661 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
      721 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
      781 tgattctgtg gataaccgta gtcggtctca atccgggagt ggcgaaggca ggggctccct
      841 tttgacgtgc ggggatgttg aagagaaccc tgggcccgga tctggaatgg tgagcaaggg
      901 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
      961 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     1021 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     1081 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     1141 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     1201 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     1261 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     1321 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     1381 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     1441 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     1501 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     1561 ccactccacc ggcggcatgg acgagctgta caagtaatgg ctgagaccag accaataaaa
     1621 aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact
     1681 ggatctatca acaggagtcc aagcgagctc gatatcaaat tacgccccgc cctgccactc
     1741 atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc
     1801 atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc
     1861 catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat caaaactggt
     1921 gaaactcacc cagggattgg ctgaaacgaa aaacatattc tcaataaacc ctttagggaa
     1981 ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta gaaactgccg
     2041 gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct catggaaaac
     2101 ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca ttgccatacg
     2161 aaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
     2221 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
     2281 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
     2341 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
     2401 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcatta
//
