LOCUS       sequence-491961-        5232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HITAG Donor plasmid for Frame 0 with 3xFLAG tag and triple Puromycin
            resistant marker (3xPuroR).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5232)
  TITLE     FLAG-tag donor plasmids for HITAG
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5232)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pDNR-3xFLAG-T2A-3xPuroR-Frame 0
FEATURES             Location/Qualifiers
     source          1..5232
                     /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             886..951
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             961..1014
                     /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             1612..1665
                     /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             2263..2316
                     /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    2940..3061
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3309..3326)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3480..4068)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3560..3579)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4268..5059,5060..5128))
                     /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 tctggatcta ctaactaagt aagctagcga atcgactcga
      841 acttcgtgtg cgagtggcgg gtctggtggc agtggagggg gatccgacta caaagaccat
      901 gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa gtccggagga
      961 gagggcagag gaagtctgct aacatgcggt gacgtagaag aaaacccagg gcctctgact
     1021 gaatacaagc ctactgtcag gttggctaca agagacgacg ttcctagagc cgtgagaact
     1081 ctggctgcag ccttcgccga ctaccccgcc acgagacaca ccgttgaccc agatcggcat
     1141 attgagagag tgactgaact gcaggagctg tttcttacaa gagttggcct cgacataggc
     1201 aaggtgtggg tggcggacga cggcgccgcc gtggccgtct ggaccactcc cgaatcagtt
     1261 gaggctggcg ccgtattcgc tgagatcggc ccgagaatgg ctgagctcag cgggagtagg
     1321 ctcgcggcac agcagcaaat ggagggactg ctggcaccac acaggcccaa agaacccgcc
     1381 tggttcctgg caaccgtcgg tgtatctccc gatcatcagg ggaaaggtct gggctctgcc
     1441 gtagtgctcc ctggcgtgga ggcagctgag agagcaggag tacctgcctt cttggagacc
     1501 tccgctccaa ggaatcttcc cttctatgaa cggttgggct tcaccgtgac agccgacgtg
     1561 gaagtccccg aaggcccccg cacttggtgc atgacgagga agcctggagc ggaaggtaga
     1621 ggttctttgc ttacctgcgg ggacgtggaa gagaacccag gcccactgac tgaatacaag
     1681 cctactgtca ggttggctac aagagacgac gttcctagag ccgtgagaac tctggctgca
     1741 gccttcgccg actaccccgc cacgagacac accgttgacc cagatcggca tattgagaga
     1801 gtgactgaac tgcaggagct gtttcttaca agagttggcc tcgacatagg caaggtgtgg
     1861 gtggcggacg acggcgccgc cgtggccgtc tggaccactc ccgaatcagt tgaggctggc
     1921 gccgtattcg ctgagatcgg cccgagaatg gctgagctca gcgggagtag gctcgcggca
     1981 cagcagcaaa tggagggact gctggcacca cacaggccca aagaacccgc ctggttcctg
     2041 gcaaccgtcg gtgtatctcc cgatcatcag gggaaaggtc tgggctctgc cgtagtgctc
     2101 cctggcgtgg aggcagctga gagagcagga gtacctgcct tcttggagac ctccgctcca
     2161 aggaatcttc ccttctatga acggttgggc ttcaccgtga cagccgacgt ggaagtcccc
     2221 gaaggccccc gcacttggtg catgacgagg aagcctggag cggagggccg agggtcactc
     2281 cttacttgtg gtgatgtcga ggagaatcct ggcccactga ctgaatacaa gcctactgtc
     2341 aggttggcta caagagacga cgttcctaga gccgtgagaa ctctggctgc agccttcgcc
     2401 gactaccccg ccacgagaca caccgttgac ccagatcggc atattgagag agtgactgaa
     2461 ctgcaggagc tgtttcttac aagagttggc ctcgacatag gcaaggtgtg ggtggcggac
     2521 gacggcgccg ccgtggccgt ctggaccact cccgaatcag ttgaggctgg cgccgtattc
     2581 gctgagatcg gcccgagaat ggctgagctc agcgggagta ggctcgcggc acagcagcaa
     2641 atggagggac tgctggcacc acacaggccc aaagaacccg cctggttcct ggcaaccgtc
     2701 ggtgtatctc ccgatcatca ggggaaaggt ctgggctctg ccgtagtgct ccctggcgtg
     2761 gaggcagctg agagagcagg agtacctgcc ttcttggaga cctccgctcc aaggaatctt
     2821 cccttctatg aacggttggg cttcaccgtg acagccgacg tggaagtccc cgaaggcccc
     2881 cgcacttggt gcatgacgag gaagcctgga gcgtgagggc ccgcttcgag cagacatgat
     2941 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     3001 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3061 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     3121 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaagatccgt cgaccgatgc
     3181 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     3241 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     3301 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3361 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3421 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3481 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3541 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3601 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3661 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3721 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3781 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3841 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3901 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3961 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4021 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4081 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4141 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4201 tcaatctaaa gtatatatga gtaaacttgg tctgacagcg gccgcaaatg ctaaaccact
     4261 gcagtggtta ccagtgcttg atcagtgagg caccgatctc agcgatctgc ctatttcgtt
     4321 cgtccatagt ggcctgactc cccgtcgtgt agatcactac gattcgtgag ggcttaccat
     4381 caggccccag cgcagcaatg atgccgcgag agccgcgttc accggccccc gatttgtcag
     4441 caatgaacca gccagcaggg agggccgagc gaagaagtgg tcctgctact ttgtccgcct
     4501 ccatccagtc tatgagctgc tgtcgtgatg ctagagtaag aagttcgcca gtgagtagtt
     4561 tccgaagagt tgtggccatt gctactggca tcgtggtatc acgctcgtcg ttcggtatgg
     4621 cttcgttcaa ctctggttcc cagcggtcaa gccgggtcac atgatcaccc atattatgaa
     4681 gaaatgcagt cagctcctta gggcctccga tcgttgtcag aagtaagttg gccgcggtgt
     4741 tgtcgctcat ggtaatggca gcactacaca attctcttac cgtcatgcca tccgtaagat
     4801 gcttttccgt gaccggcgag tactcaacca agtcgttttg tgagtagtgt atacggcgac
     4861 caagctgctc ttgcccggcg tctatacggg acaacaccgc gccacatagc agtactttga
     4921 aagtgctcat catcgggaat cgttcttcgg ggcggaaaga ctcaaggatc ttgccgctat
     4981 tgagatccag ttcgatatag cccactcttg cacccagttg atcttcagca tcttttactt
     5041 tcaccagcgt ttcggggtgt gcaaaaacag gcaagcaaaa tgccgcaaag aagggaatga
     5101 gtgcgacacg aaaatgttgg atgctcatac tcgtcctttt tcaatattat tgaagcattt
     5161 atcagggtta ctagtacgtc tctcaaggat aagtaagtaa tattaaggta cgggaggtat
     5221 tggacaggcc gc
//
