LOCUS       sequence-491966-        5384 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HITAG Donor plasmid for Frame 2 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 5384)
  TITLE     FLAG-tag donor plasmids for HITAG
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5384)
  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            "Unpublished"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 2
FEATURES             Location/Qualifiers
     source          1..5384
                     /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             888..953
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             963..1028
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1038..1103
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1113..1166
                     /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             1764..1817
                     /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             2415..2468
                     /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    3092..3213
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3461..3478)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3632..4220)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3712..3731)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4420..5211,5212..5280))
                     /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 cgagtctggc gggtctggtg gcagtggagg gggatccgac tacaaagacc
      901 atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac aagggaagat
      961 ccgactacaa agaccatgac ggtgattata aagatcatga catcgattac aaggatgacg
     1021 atgacaaggg aagatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     1081 attacaagga tgacgatgac aagtccggag gagagggcag aggaagtctg ctaacatgcg
     1141 gtgacgtaga agaaaaccca gggcctctga ctgaatacaa gcctactgtc aggttggcta
     1201 caagagacga cgttcctaga gccgtgagaa ctctggctgc agccttcgcc gactaccccg
     1261 ccacgagaca caccgttgac ccagatcggc atattgagag agtgactgaa ctgcaggagc
     1321 tgtttcttac aagagttggc ctcgacatag gcaaggtgtg ggtggcggac gacggcgccg
     1381 ccgtggccgt ctggaccact cccgaatcag ttgaggctgg cgccgtattc gctgagatcg
     1441 gcccgagaat ggctgagctc agcgggagta ggctcgcggc acagcagcaa atggagggac
     1501 tgctggcacc acacaggccc aaagaacccg cctggttcct ggcaaccgtc ggtgtatctc
     1561 ccgatcatca ggggaaaggt ctgggctctg ccgtagtgct ccctggcgtg gaggcagctg
     1621 agagagcagg agtacctgcc ttcttggaga cctccgctcc aaggaatctt cccttctatg
     1681 aacggttggg cttcaccgtg acagccgacg tggaagtccc cgaaggcccc cgcacttggt
     1741 gcatgacgag gaagcctgga gcggaaggta gaggttcttt gcttacctgc ggggacgtgg
     1801 aagagaaccc aggcccactg actgaataca agcctactgt caggttggct acaagagacg
     1861 acgttcctag agccgtgaga actctggctg cagccttcgc cgactacccc gccacgagac
     1921 acaccgttga cccagatcgg catattgaga gagtgactga actgcaggag ctgtttctta
     1981 caagagttgg cctcgacata ggcaaggtgt gggtggcgga cgacggcgcc gccgtggccg
     2041 tctggaccac tcccgaatca gttgaggctg gcgccgtatt cgctgagatc ggcccgagaa
     2101 tggctgagct cagcgggagt aggctcgcgg cacagcagca aatggaggga ctgctggcac
     2161 cacacaggcc caaagaaccc gcctggttcc tggcaaccgt cggtgtatct cccgatcatc
     2221 aggggaaagg tctgggctct gccgtagtgc tccctggcgt ggaggcagct gagagagcag
     2281 gagtacctgc cttcttggag acctccgctc caaggaatct tcccttctat gaacggttgg
     2341 gcttcaccgt gacagccgac gtggaagtcc ccgaaggccc ccgcacttgg tgcatgacga
     2401 ggaagcctgg agcggagggc cgagggtcac tccttacttg tggtgatgtc gaggagaatc
     2461 ctggcccact gactgaatac aagcctactg tcaggttggc tacaagagac gacgttccta
     2521 gagccgtgag aactctggct gcagccttcg ccgactaccc cgccacgaga cacaccgttg
     2581 acccagatcg gcatattgag agagtgactg aactgcagga gctgtttctt acaagagttg
     2641 gcctcgacat aggcaaggtg tgggtggcgg acgacggcgc cgccgtggcc gtctggacca
     2701 ctcccgaatc agttgaggct ggcgccgtat tcgctgagat cggcccgaga atggctgagc
     2761 tcagcgggag taggctcgcg gcacagcagc aaatggaggg actgctggca ccacacaggc
     2821 ccaaagaacc cgcctggttc ctggcaaccg tcggtgtatc tcccgatcat caggggaaag
     2881 gtctgggctc tgccgtagtg ctccctggcg tggaggcagc tgagagagca ggagtacctg
     2941 ccttcttgga gacctccgct ccaaggaatc ttcccttcta tgaacggttg ggcttcaccg
     3001 tgacagccga cgtggaagtc cccgaaggcc cccgcacttg gtgcatgacg aggaagcctg
     3061 gagcgtgagg gcccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     3121 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     3181 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     3241 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     3301 ggtaaaatcg ataaagatcc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc
     3361 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     3421 gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc
     3481 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3541 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3601 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3661 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3721 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3781 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3841 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3901 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3961 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4021 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4081 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4141 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4201 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4261 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4321 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4381 ggtctgacag cggccgcaaa tgctaaacca ctgcagtggt taccagtgct tgatcagtga
     4441 ggcaccgatc tcagcgatct gcctatttcg ttcgtccata gtggcctgac tccccgtcgt
     4501 gtagatcact acgattcgtg agggcttacc atcaggcccc agcgcagcaa tgatgccgcg
     4561 agagccgcgt tcaccggccc ccgatttgtc agcaatgaac cagccagcag ggagggccga
     4621 gcgaagaagt ggtcctgcta ctttgtccgc ctccatccag tctatgagct gctgtcgtga
     4681 tgctagagta agaagttcgc cagtgagtag tttccgaaga gttgtggcca ttgctactgg
     4741 catcgtggta tcacgctcgt cgttcggtat ggcttcgttc aactctggtt cccagcggtc
     4801 aagccgggtc acatgatcac ccatattatg aagaaatgca gtcagctcct tagggcctcc
     4861 gatcgttgtc agaagtaagt tggccgcggt gttgtcgctc atggtaatgg cagcactaca
     4921 caattctctt accgtcatgc catccgtaag atgcttttcc gtgaccggcg agtactcaac
     4981 caagtcgttt tgtgagtagt gtatacggcg accaagctgc tcttgcccgg cgtctatacg
     5041 ggacaacacc gcgccacata gcagtacttt gaaagtgctc atcatcggga atcgttcttc
     5101 ggggcggaaa gactcaagga tcttgccgct attgagatcc agttcgatat agcccactct
     5161 tgcacccagt tgatcttcag catcttttac tttcaccagc gtttcggggt gtgcaaaaac
     5221 aggcaagcaa aatgccgcaa agaagggaat gagtgcgaca cgaaaatgtt ggatgctcat
     5281 actcgtcctt tttcaatatt attgaagcat ttatcagggt tactagtacg tctctcaagg
     5341 ataagtaagt aatattaagg tacgggaggt attggacagg ccgc
//
