LOCUS       sequence-410549-        5893 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid for Frame 1 with mCherry tag and triple Puromycin 
            resistant marker (3xPuroR).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5893)
  AUTHORS   Kim J, Kratz AF, Chen S, Sheng J, Kim HK, Zhang L, Singh BK, Chavez 
            A
  TITLE     High-throughput tagging of endogenous loci for rapid 
            characterization of protein function.
  JOURNAL   Sci Adv. 2024 May 3;10(18):eadg8771. doi: 10.1126/sciadv.adg8771. 
            Epub 2024 May 1.
  PUBMED    38691600
REFERENCE   2  (bases 1 to 5893)
  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: "Sci Adv."; 
            date: "2024-05-3"; volume: "10(18):eadg8771. doi"; pages: " 
            10.1126/sciadv.adg8771. Epub 2024 May 1"
COMMENT     Sequence Label: pDNR-mCherry-T2A-3xPuroR-Frame 1
FEATURES             Location/Qualifiers
     source          1..5893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    166..300
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    420..554
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    674..808
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             908..1612
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1050..1068)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1318..1337
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1622..1675
                     /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             2273..2326
                     /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             2924..2977
                     /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"
     polyA_signal    3601..3722
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3970..3987)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4141..4729)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4221..4240)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4929..5720,5721..5789))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 cgcgatgaat aaatgaaagc ttgcagatct gcgactctag aggatctgcg actctagagg
       61 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
      121 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
      181 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
      241 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
      301 tgcgactcta gaggatcata atcagccata ccacatttgt agaggtttta cttgctttaa
      361 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
      421 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
      481 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
      541 atcatgtctg gatctgcgac tctagaggat cataatcagc cataccacat ttgtagaggt
      601 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
      661 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
      721 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
      781 catcaatgta tcttatcatg tctggatccc catcaagctg atccggaact aactaagtaa
      841 gctagcgaat cgactcgaac ttcgtgtgcg agtcggcggg tctggtggca gtggaggggg
      901 atccctggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
      961 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
     1021 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
     1081 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
     1141 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
     1201 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
     1261 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     1321 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
     1381 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     1441 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
     1501 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     1561 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtccggagg
     1621 agagggcaga ggaagtctgc taacatgcgg tgacgtagaa gaaaacccag ggcctctgac
     1681 tgaatacaag cctactgtca ggttggctac aagagacgac gttcctagag ccgtgagaac
     1741 tctggctgca gccttcgccg actaccccgc cacgagacac accgttgacc cagatcggca
     1801 tattgagaga gtgactgaac tgcaggagct gtttcttaca agagttggcc tcgacatagg
     1861 caaggtgtgg gtggcggacg acggcgccgc cgtggccgtc tggaccactc ccgaatcagt
     1921 tgaggctggc gccgtattcg ctgagatcgg cccgagaatg gctgagctca gcgggagtag
     1981 gctcgcggca cagcagcaaa tggagggact gctggcacca cacaggccca aagaacccgc
     2041 ctggttcctg gcaaccgtcg gtgtatctcc cgatcatcag gggaaaggtc tgggctctgc
     2101 cgtagtgctc cctggcgtgg aggcagctga gagagcagga gtacctgcct tcttggagac
     2161 ctccgctcca aggaatcttc ccttctatga acggttgggc ttcaccgtga cagccgacgt
     2221 ggaagtcccc gaaggccccc gcacttggtg catgacgagg aagcctggag cggaaggtag
     2281 aggttctttg cttacctgcg gggacgtgga agagaaccca ggcccactga ctgaatacaa
     2341 gcctactgtc aggttggcta caagagacga cgttcctaga gccgtgagaa ctctggctgc
     2401 agccttcgcc gactaccccg ccacgagaca caccgttgac ccagatcggc atattgagag
     2461 agtgactgaa ctgcaggagc tgtttcttac aagagttggc ctcgacatag gcaaggtgtg
     2521 ggtggcggac gacggcgccg ccgtggccgt ctggaccact cccgaatcag ttgaggctgg
     2581 cgccgtattc gctgagatcg gcccgagaat ggctgagctc agcgggagta ggctcgcggc
     2641 acagcagcaa atggagggac tgctggcacc acacaggccc aaagaacccg cctggttcct
     2701 ggcaaccgtc ggtgtatctc ccgatcatca ggggaaaggt ctgggctctg ccgtagtgct
     2761 ccctggcgtg gaggcagctg agagagcagg agtacctgcc ttcttggaga cctccgctcc
     2821 aaggaatctt cccttctatg aacggttggg cttcaccgtg acagccgacg tggaagtccc
     2881 cgaaggcccc cgcacttggt gcatgacgag gaagcctgga gcggagggcc gagggtcact
     2941 ccttacttgt ggtgatgtcg aggagaatcc tggcccactg actgaataca agcctactgt
     3001 caggttggct acaagagacg acgttcctag agccgtgaga actctggctg cagccttcgc
     3061 cgactacccc gccacgagac acaccgttga cccagatcgg catattgaga gagtgactga
     3121 actgcaggag ctgtttctta caagagttgg cctcgacata ggcaaggtgt gggtggcgga
     3181 cgacggcgcc gccgtggccg tctggaccac tcccgaatca gttgaggctg gcgccgtatt
     3241 cgctgagatc ggcccgagaa tggctgagct cagcgggagt aggctcgcgg cacagcagca
     3301 aatggaggga ctgctggcac cacacaggcc caaagaaccc gcctggttcc tggcaaccgt
     3361 cggtgtatct cccgatcatc aggggaaagg tctgggctct gccgtagtgc tccctggcgt
     3421 ggaggcagct gagagagcag gagtacctgc cttcttggag acctccgctc caaggaatct
     3481 tcccttctat gaacggttgg gcttcaccgt gacagccgac gtggaagtcc ccgaaggccc
     3541 ccgcacttgg tgcatgacga ggaagcctgg agcgtgaggg cccgcttcga gcagacatga
     3601 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     3661 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     3721 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     3781 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaagatccg tcgaccgatg
     3841 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc
     3901 gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc
     3961 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4021 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4081 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4141 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4201 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4261 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4321 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4381 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4441 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4501 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4561 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4621 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4681 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4741 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4801 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4861 atcaatctaa agtatatatg agtaaacttg gtctgacagc ggccgcaaat gctaaaccac
     4921 tgcagtggtt accagtgctt gatcagtgag gcaccgatct cagcgatctg cctatttcgt
     4981 tcgtccatag tggcctgact ccccgtcgtg tagatcacta cgattcgtga gggcttacca
     5041 tcaggcccca gcgcagcaat gatgccgcga gagccgcgtt caccggcccc cgatttgtca
     5101 gcaatgaacc agccagcagg gagggccgag cgaagaagtg gtcctgctac tttgtccgcc
     5161 tccatccagt ctatgagctg ctgtcgtgat gctagagtaa gaagttcgcc agtgagtagt
     5221 ttccgaagag ttgtggccat tgctactggc atcgtggtat cacgctcgtc gttcggtatg
     5281 gcttcgttca actctggttc ccagcggtca agccgggtca catgatcacc catattatga
     5341 agaaatgcag tcagctcctt agggcctccg atcgttgtca gaagtaagtt ggccgcggtg
     5401 ttgtcgctca tggtaatggc agcactacac aattctctta ccgtcatgcc atccgtaaga
     5461 tgcttttccg tgaccggcga gtactcaacc aagtcgtttt gtgagtagtg tatacggcga
     5521 ccaagctgct cttgcccggc gtctatacgg gacaacaccg cgccacatag cagtactttg
     5581 aaagtgctca tcatcgggaa tcgttcttcg gggcggaaag actcaaggat cttgccgcta
     5641 ttgagatcca gttcgatata gcccactctt gcacccagtt gatcttcagc atcttttact
     5701 ttcaccagcg tttcggggtg tgcaaaaaca ggcaagcaaa atgccgcaaa gaagggaatg
     5761 agtgcgacac gaaaatgttg gatgctcata ctcgtccttt ttcaatatta ttgaagcatt
     5821 tatcagggtt actagtacgt ctctcaagga taagtaagta atattaaggt acgggaggta
     5881 ttggacaggc cgc
//
