LOCUS       sequence-491963-        5233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HITAG Donor plasmid for Frame 1 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 5233)
  TITLE     FLAG-tag donor plasmids for HITAG
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5233)
  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            "Unpublished"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 1
FEATURES             Location/Qualifiers
     source          1..5233
                     /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             887..952
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             962..1015
                     /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             1613..1666
                     /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             2264..2317
                     /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    2941..3062
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3310..3327)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3481..4069)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3561..3580)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4269..5060,5061..5129))
                     /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 cgagtcggcg ggtctggtgg cagtggaggg ggatccgact acaaagacca
      901 tgacggtgat tataaagatc atgacatcga ttacaaggat gacgatgaca agtccggagg
      961 agagggcaga ggaagtctgc taacatgcgg tgacgtagaa gaaaacccag ggcctctgac
     1021 tgaatacaag cctactgtca ggttggctac aagagacgac gttcctagag ccgtgagaac
     1081 tctggctgca gccttcgccg actaccccgc cacgagacac accgttgacc cagatcggca
     1141 tattgagaga gtgactgaac tgcaggagct gtttcttaca agagttggcc tcgacatagg
     1201 caaggtgtgg gtggcggacg acggcgccgc cgtggccgtc tggaccactc ccgaatcagt
     1261 tgaggctggc gccgtattcg ctgagatcgg cccgagaatg gctgagctca gcgggagtag
     1321 gctcgcggca cagcagcaaa tggagggact gctggcacca cacaggccca aagaacccgc
     1381 ctggttcctg gcaaccgtcg gtgtatctcc cgatcatcag gggaaaggtc tgggctctgc
     1441 cgtagtgctc cctggcgtgg aggcagctga gagagcagga gtacctgcct tcttggagac
     1501 ctccgctcca aggaatcttc ccttctatga acggttgggc ttcaccgtga cagccgacgt
     1561 ggaagtcccc gaaggccccc gcacttggtg catgacgagg aagcctggag cggaaggtag
     1621 aggttctttg cttacctgcg gggacgtgga agagaaccca ggcccactga ctgaatacaa
     1681 gcctactgtc aggttggcta caagagacga cgttcctaga gccgtgagaa ctctggctgc
     1741 agccttcgcc gactaccccg ccacgagaca caccgttgac ccagatcggc atattgagag
     1801 agtgactgaa ctgcaggagc tgtttcttac aagagttggc ctcgacatag gcaaggtgtg
     1861 ggtggcggac gacggcgccg ccgtggccgt ctggaccact cccgaatcag ttgaggctgg
     1921 cgccgtattc gctgagatcg gcccgagaat ggctgagctc agcgggagta ggctcgcggc
     1981 acagcagcaa atggagggac tgctggcacc acacaggccc aaagaacccg cctggttcct
     2041 ggcaaccgtc ggtgtatctc ccgatcatca ggggaaaggt ctgggctctg ccgtagtgct
     2101 ccctggcgtg gaggcagctg agagagcagg agtacctgcc ttcttggaga cctccgctcc
     2161 aaggaatctt cccttctatg aacggttggg cttcaccgtg acagccgacg tggaagtccc
     2221 cgaaggcccc cgcacttggt gcatgacgag gaagcctgga gcggagggcc gagggtcact
     2281 ccttacttgt ggtgatgtcg aggagaatcc tggcccactg actgaataca agcctactgt
     2341 caggttggct acaagagacg acgttcctag agccgtgaga actctggctg cagccttcgc
     2401 cgactacccc gccacgagac acaccgttga cccagatcgg catattgaga gagtgactga
     2461 actgcaggag ctgtttctta caagagttgg cctcgacata ggcaaggtgt gggtggcgga
     2521 cgacggcgcc gccgtggccg tctggaccac tcccgaatca gttgaggctg gcgccgtatt
     2581 cgctgagatc ggcccgagaa tggctgagct cagcgggagt aggctcgcgg cacagcagca
     2641 aatggaggga ctgctggcac cacacaggcc caaagaaccc gcctggttcc tggcaaccgt
     2701 cggtgtatct cccgatcatc aggggaaagg tctgggctct gccgtagtgc tccctggcgt
     2761 ggaggcagct gagagagcag gagtacctgc cttcttggag acctccgctc caaggaatct
     2821 tcccttctat gaacggttgg gcttcaccgt gacagccgac gtggaagtcc ccgaaggccc
     2881 ccgcacttgg tgcatgacga ggaagcctgg agcgtgaggg cccgcttcga gcagacatga
     2941 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     3001 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     3061 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     3121 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaagatccg tcgaccgatg
     3181 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc
     3241 gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc
     3301 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3361 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3421 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3481 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3541 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3601 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3661 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3721 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3781 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3841 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3901 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3961 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4021 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4081 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4141 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4201 atcaatctaa agtatatatg agtaaacttg gtctgacagc ggccgcaaat gctaaaccac
     4261 tgcagtggtt accagtgctt gatcagtgag gcaccgatct cagcgatctg cctatttcgt
     4321 tcgtccatag tggcctgact ccccgtcgtg tagatcacta cgattcgtga gggcttacca
     4381 tcaggcccca gcgcagcaat gatgccgcga gagccgcgtt caccggcccc cgatttgtca
     4441 gcaatgaacc agccagcagg gagggccgag cgaagaagtg gtcctgctac tttgtccgcc
     4501 tccatccagt ctatgagctg ctgtcgtgat gctagagtaa gaagttcgcc agtgagtagt
     4561 ttccgaagag ttgtggccat tgctactggc atcgtggtat cacgctcgtc gttcggtatg
     4621 gcttcgttca actctggttc ccagcggtca agccgggtca catgatcacc catattatga
     4681 agaaatgcag tcagctcctt agggcctccg atcgttgtca gaagtaagtt ggccgcggtg
     4741 ttgtcgctca tggtaatggc agcactacac aattctctta ccgtcatgcc atccgtaaga
     4801 tgcttttccg tgaccggcga gtactcaacc aagtcgtttt gtgagtagtg tatacggcga
     4861 ccaagctgct cttgcccggc gtctatacgg gacaacaccg cgccacatag cagtactttg
     4921 aaagtgctca tcatcgggaa tcgttcttcg gggcggaaag actcaaggat cttgccgcta
     4981 ttgagatcca gttcgatata gcccactctt gcacccagtt gatcttcagc atcttttact
     5041 ttcaccagcg tttcggggtg tgcaaaaaca ggcaagcaaa atgccgcaaa gaagggaatg
     5101 agtgcgacac gaaaatgttg gatgctcata ctcgtccttt ttcaatatta ttgaagcatt
     5161 tatcagggtt actagtacgt ctctcaagga taagtaagta atattaaggt acgggaggta
     5221 ttggacaggc cgc
//
