LOCUS       sequence-491941-        5383 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HITAG Donor plasmid for Frame 1 with 9xFLAG tag and triple Puromycin
            resistant marker (3xPuroR).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5383)
  TITLE     FLAG-tag donor plasmids for HITAG
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5383)
  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            "Unpublished"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-9xFLAG-T2A-3xPuroR-Frame 1
FEATURES             Location/Qualifiers
     source          1..5383
                     /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..1027
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1037..1102
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1112..1165
                     /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             1763..1816
                     /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             2414..2467
                     /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    3091..3212
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3460..3477)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3631..4219)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3711..3730)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4419..5210,5211..5279))
                     /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 agggaagatc
      961 cgactacaaa gaccatgacg gtgattataa agatcatgac atcgattaca aggatgacga
     1021 tgacaaggga agatccgact acaaagacca tgacggtgat tataaagatc atgacatcga
     1081 ttacaaggat gacgatgaca agtccggagg agagggcaga ggaagtctgc taacatgcgg
     1141 tgacgtagaa gaaaacccag ggcctctgac tgaatacaag cctactgtca ggttggctac
     1201 aagagacgac gttcctagag ccgtgagaac tctggctgca gccttcgccg actaccccgc
     1261 cacgagacac accgttgacc cagatcggca tattgagaga gtgactgaac tgcaggagct
     1321 gtttcttaca agagttggcc tcgacatagg caaggtgtgg gtggcggacg acggcgccgc
     1381 cgtggccgtc tggaccactc ccgaatcagt tgaggctggc gccgtattcg ctgagatcgg
     1441 cccgagaatg gctgagctca gcgggagtag gctcgcggca cagcagcaaa tggagggact
     1501 gctggcacca cacaggccca aagaacccgc ctggttcctg gcaaccgtcg gtgtatctcc
     1561 cgatcatcag gggaaaggtc tgggctctgc cgtagtgctc cctggcgtgg aggcagctga
     1621 gagagcagga gtacctgcct tcttggagac ctccgctcca aggaatcttc ccttctatga
     1681 acggttgggc ttcaccgtga cagccgacgt ggaagtcccc gaaggccccc gcacttggtg
     1741 catgacgagg aagcctggag cggaaggtag aggttctttg cttacctgcg gggacgtgga
     1801 agagaaccca ggcccactga ctgaatacaa gcctactgtc aggttggcta caagagacga
     1861 cgttcctaga gccgtgagaa ctctggctgc agccttcgcc gactaccccg ccacgagaca
     1921 caccgttgac ccagatcggc atattgagag agtgactgaa ctgcaggagc tgtttcttac
     1981 aagagttggc ctcgacatag gcaaggtgtg ggtggcggac gacggcgccg ccgtggccgt
     2041 ctggaccact cccgaatcag ttgaggctgg cgccgtattc gctgagatcg gcccgagaat
     2101 ggctgagctc agcgggagta ggctcgcggc acagcagcaa atggagggac tgctggcacc
     2161 acacaggccc aaagaacccg cctggttcct ggcaaccgtc ggtgtatctc ccgatcatca
     2221 ggggaaaggt ctgggctctg ccgtagtgct ccctggcgtg gaggcagctg agagagcagg
     2281 agtacctgcc ttcttggaga cctccgctcc aaggaatctt cccttctatg aacggttggg
     2341 cttcaccgtg acagccgacg tggaagtccc cgaaggcccc cgcacttggt gcatgacgag
     2401 gaagcctgga gcggagggcc gagggtcact ccttacttgt ggtgatgtcg aggagaatcc
     2461 tggcccactg actgaataca agcctactgt caggttggct acaagagacg acgttcctag
     2521 agccgtgaga actctggctg cagccttcgc cgactacccc gccacgagac acaccgttga
     2581 cccagatcgg catattgaga gagtgactga actgcaggag ctgtttctta caagagttgg
     2641 cctcgacata ggcaaggtgt gggtggcgga cgacggcgcc gccgtggccg tctggaccac
     2701 tcccgaatca gttgaggctg gcgccgtatt cgctgagatc ggcccgagaa tggctgagct
     2761 cagcgggagt aggctcgcgg cacagcagca aatggaggga ctgctggcac cacacaggcc
     2821 caaagaaccc gcctggttcc tggcaaccgt cggtgtatct cccgatcatc aggggaaagg
     2881 tctgggctct gccgtagtgc tccctggcgt ggaggcagct gagagagcag gagtacctgc
     2941 cttcttggag acctccgctc caaggaatct tcccttctat gaacggttgg gcttcaccgt
     3001 gacagccgac gtggaagtcc ccgaaggccc ccgcacttgg tgcatgacga ggaagcctgg
     3061 agcgtgaggg cccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     3121 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3181 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     3241 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     3301 gtaaaatcga taaagatccg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     3361 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     3421 caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg
     3481 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3541 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3601 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3661 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3721 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3781 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3841 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3901 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3961 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4021 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4081 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4141 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4201 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4261 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4321 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4381 gtctgacagc ggccgcaaat gctaaaccac tgcagtggtt accagtgctt gatcagtgag
     4441 gcaccgatct cagcgatctg cctatttcgt tcgtccatag tggcctgact ccccgtcgtg
     4501 tagatcacta cgattcgtga gggcttacca tcaggcccca gcgcagcaat gatgccgcga
     4561 gagccgcgtt caccggcccc cgatttgtca gcaatgaacc agccagcagg gagggccgag
     4621 cgaagaagtg gtcctgctac tttgtccgcc tccatccagt ctatgagctg ctgtcgtgat
     4681 gctagagtaa gaagttcgcc agtgagtagt ttccgaagag ttgtggccat tgctactggc
     4741 atcgtggtat cacgctcgtc gttcggtatg gcttcgttca actctggttc ccagcggtca
     4801 agccgggtca catgatcacc catattatga agaaatgcag tcagctcctt agggcctccg
     4861 atcgttgtca gaagtaagtt ggccgcggtg ttgtcgctca tggtaatggc agcactacac
     4921 aattctctta ccgtcatgcc atccgtaaga tgcttttccg tgaccggcga gtactcaacc
     4981 aagtcgtttt gtgagtagtg tatacggcga ccaagctgct cttgcccggc gtctatacgg
     5041 gacaacaccg cgccacatag cagtactttg aaagtgctca tcatcgggaa tcgttcttcg
     5101 gggcggaaag actcaaggat cttgccgcta ttgagatcca gttcgatata gcccactctt
     5161 gcacccagtt gatcttcagc atcttttact ttcaccagcg tttcggggtg tgcaaaaaca
     5221 ggcaagcaaa atgccgcaaa gaagggaatg agtgcgacac gaaaatgttg gatgctcata
     5281 ctcgtccttt ttcaatatta ttgaagcatt tatcagggtt actagtacgt ctctcaagga
     5341 taagtaagta atattaaggt acgggaggta ttggacaggc cgc
//
