LOCUS       sequence-491964-        5234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HITAG Donor plasmid for Frame 2 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 5234)
  TITLE     FLAG-tag donor plasmids for HITAG
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5234)
  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            "Unpublished"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 2
FEATURES             Location/Qualifiers
     source          1..5234
                     /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..1016
                     /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             1614..1667
                     /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             2265..2318
                     /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    2942..3063
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3311..3328)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3482..4070)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3562..3581)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4270..5061,5062..5130))
                     /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 aagtccggag
      961 gagagggcag aggaagtctg ctaacatgcg gtgacgtaga agaaaaccca gggcctctga
     1021 ctgaatacaa gcctactgtc aggttggcta caagagacga cgttcctaga gccgtgagaa
     1081 ctctggctgc agccttcgcc gactaccccg ccacgagaca caccgttgac ccagatcggc
     1141 atattgagag agtgactgaa ctgcaggagc tgtttcttac aagagttggc ctcgacatag
     1201 gcaaggtgtg ggtggcggac gacggcgccg ccgtggccgt ctggaccact cccgaatcag
     1261 ttgaggctgg cgccgtattc gctgagatcg gcccgagaat ggctgagctc agcgggagta
     1321 ggctcgcggc acagcagcaa atggagggac tgctggcacc acacaggccc aaagaacccg
     1381 cctggttcct ggcaaccgtc ggtgtatctc ccgatcatca ggggaaaggt ctgggctctg
     1441 ccgtagtgct ccctggcgtg gaggcagctg agagagcagg agtacctgcc ttcttggaga
     1501 cctccgctcc aaggaatctt cccttctatg aacggttggg cttcaccgtg acagccgacg
     1561 tggaagtccc cgaaggcccc cgcacttggt gcatgacgag gaagcctgga gcggaaggta
     1621 gaggttcttt gcttacctgc ggggacgtgg aagagaaccc aggcccactg actgaataca
     1681 agcctactgt caggttggct acaagagacg acgttcctag agccgtgaga actctggctg
     1741 cagccttcgc cgactacccc gccacgagac acaccgttga cccagatcgg catattgaga
     1801 gagtgactga actgcaggag ctgtttctta caagagttgg cctcgacata ggcaaggtgt
     1861 gggtggcgga cgacggcgcc gccgtggccg tctggaccac tcccgaatca gttgaggctg
     1921 gcgccgtatt cgctgagatc ggcccgagaa tggctgagct cagcgggagt aggctcgcgg
     1981 cacagcagca aatggaggga ctgctggcac cacacaggcc caaagaaccc gcctggttcc
     2041 tggcaaccgt cggtgtatct cccgatcatc aggggaaagg tctgggctct gccgtagtgc
     2101 tccctggcgt ggaggcagct gagagagcag gagtacctgc cttcttggag acctccgctc
     2161 caaggaatct tcccttctat gaacggttgg gcttcaccgt gacagccgac gtggaagtcc
     2221 ccgaaggccc ccgcacttgg tgcatgacga ggaagcctgg agcggagggc cgagggtcac
     2281 tccttacttg tggtgatgtc gaggagaatc ctggcccact gactgaatac aagcctactg
     2341 tcaggttggc tacaagagac gacgttccta gagccgtgag aactctggct gcagccttcg
     2401 ccgactaccc cgccacgaga cacaccgttg acccagatcg gcatattgag agagtgactg
     2461 aactgcagga gctgtttctt acaagagttg gcctcgacat aggcaaggtg tgggtggcgg
     2521 acgacggcgc cgccgtggcc gtctggacca ctcccgaatc agttgaggct ggcgccgtat
     2581 tcgctgagat cggcccgaga atggctgagc tcagcgggag taggctcgcg gcacagcagc
     2641 aaatggaggg actgctggca ccacacaggc ccaaagaacc cgcctggttc ctggcaaccg
     2701 tcggtgtatc tcccgatcat caggggaaag gtctgggctc tgccgtagtg ctccctggcg
     2761 tggaggcagc tgagagagca ggagtacctg ccttcttgga gacctccgct ccaaggaatc
     2821 ttcccttcta tgaacggttg ggcttcaccg tgacagccga cgtggaagtc cccgaaggcc
     2881 cccgcacttg gtgcatgacg aggaagcctg gagcgtgagg gcccgcttcg agcagacatg
     2941 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     3001 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     3061 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     3121 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaagatcc gtcgaccgat
     3181 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     3241 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     3301 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3361 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3421 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3481 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3541 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3601 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3661 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3721 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3781 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3841 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3901 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3961 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4021 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4081 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4141 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4201 aatcaatcta aagtatatat gagtaaactt ggtctgacag cggccgcaaa tgctaaacca
     4261 ctgcagtggt taccagtgct tgatcagtga ggcaccgatc tcagcgatct gcctatttcg
     4321 ttcgtccata gtggcctgac tccccgtcgt gtagatcact acgattcgtg agggcttacc
     4381 atcaggcccc agcgcagcaa tgatgccgcg agagccgcgt tcaccggccc ccgatttgtc
     4441 agcaatgaac cagccagcag ggagggccga gcgaagaagt ggtcctgcta ctttgtccgc
     4501 ctccatccag tctatgagct gctgtcgtga tgctagagta agaagttcgc cagtgagtag
     4561 tttccgaaga gttgtggcca ttgctactgg catcgtggta tcacgctcgt cgttcggtat
     4621 ggcttcgttc aactctggtt cccagcggtc aagccgggtc acatgatcac ccatattatg
     4681 aagaaatgca gtcagctcct tagggcctcc gatcgttgtc agaagtaagt tggccgcggt
     4741 gttgtcgctc atggtaatgg cagcactaca caattctctt accgtcatgc catccgtaag
     4801 atgcttttcc gtgaccggcg agtactcaac caagtcgttt tgtgagtagt gtatacggcg
     4861 accaagctgc tcttgcccgg cgtctatacg ggacaacacc gcgccacata gcagtacttt
     4921 gaaagtgctc atcatcggga atcgttcttc ggggcggaaa gactcaagga tcttgccgct
     4981 attgagatcc agttcgatat agcccactct tgcacccagt tgatcttcag catcttttac
     5041 tttcaccagc gtttcggggt gtgcaaaaac aggcaagcaa aatgccgcaa agaagggaat
     5101 gagtgcgaca cgaaaatgtt ggatgctcat actcgtcctt tttcaatatt attgaagcat
     5161 ttatcagggt tactagtacg tctctcaagg ataagtaagt aatattaagg tacgggaggt
     5221 attggacagg ccgc
//
