LOCUS       sequence-491944-        5382 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HITAG Donor plasmid for Frame 0 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 5382)
  TITLE     FLAG-tag donor plasmids for HITAG
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5382)
  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            "Unpublished"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 0
FEATURES             Location/Qualifiers
     source          1..5382
                     /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..1026
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1036..1101
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1111..1164
                     /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             1762..1815
                     /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             2413..2466
                     /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    3090..3211
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3459..3476)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3630..4218)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3710..3729)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4418..5209,5210..5278))
                     /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 gggaagatcc
      961 gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat
     1021 gacaagggaa gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgat
     1081 tacaaggatg acgatgacaa gtccggagga gagggcagag gaagtctgct aacatgcggt
     1141 gacgtagaag aaaacccagg gcctctgact gaatacaagc ctactgtcag gttggctaca
     1201 agagacgacg ttcctagagc cgtgagaact ctggctgcag ccttcgccga ctaccccgcc
     1261 acgagacaca ccgttgaccc agatcggcat attgagagag tgactgaact gcaggagctg
     1321 tttcttacaa gagttggcct cgacataggc aaggtgtggg tggcggacga cggcgccgcc
     1381 gtggccgtct ggaccactcc cgaatcagtt gaggctggcg ccgtattcgc tgagatcggc
     1441 ccgagaatgg ctgagctcag cgggagtagg ctcgcggcac agcagcaaat ggagggactg
     1501 ctggcaccac acaggcccaa agaacccgcc tggttcctgg caaccgtcgg tgtatctccc
     1561 gatcatcagg ggaaaggtct gggctctgcc gtagtgctcc ctggcgtgga ggcagctgag
     1621 agagcaggag tacctgcctt cttggagacc tccgctccaa ggaatcttcc cttctatgaa
     1681 cggttgggct tcaccgtgac agccgacgtg gaagtccccg aaggcccccg cacttggtgc
     1741 atgacgagga agcctggagc ggaaggtaga ggttctttgc ttacctgcgg ggacgtggaa
     1801 gagaacccag gcccactgac tgaatacaag cctactgtca ggttggctac aagagacgac
     1861 gttcctagag ccgtgagaac tctggctgca gccttcgccg actaccccgc cacgagacac
     1921 accgttgacc cagatcggca tattgagaga gtgactgaac tgcaggagct gtttcttaca
     1981 agagttggcc tcgacatagg caaggtgtgg gtggcggacg acggcgccgc cgtggccgtc
     2041 tggaccactc ccgaatcagt tgaggctggc gccgtattcg ctgagatcgg cccgagaatg
     2101 gctgagctca gcgggagtag gctcgcggca cagcagcaaa tggagggact gctggcacca
     2161 cacaggccca aagaacccgc ctggttcctg gcaaccgtcg gtgtatctcc cgatcatcag
     2221 gggaaaggtc tgggctctgc cgtagtgctc cctggcgtgg aggcagctga gagagcagga
     2281 gtacctgcct tcttggagac ctccgctcca aggaatcttc ccttctatga acggttgggc
     2341 ttcaccgtga cagccgacgt ggaagtcccc gaaggccccc gcacttggtg catgacgagg
     2401 aagcctggag cggagggccg agggtcactc cttacttgtg gtgatgtcga ggagaatcct
     2461 ggcccactga ctgaatacaa gcctactgtc aggttggcta caagagacga cgttcctaga
     2521 gccgtgagaa ctctggctgc agccttcgcc gactaccccg ccacgagaca caccgttgac
     2581 ccagatcggc atattgagag agtgactgaa ctgcaggagc tgtttcttac aagagttggc
     2641 ctcgacatag gcaaggtgtg ggtggcggac gacggcgccg ccgtggccgt ctggaccact
     2701 cccgaatcag ttgaggctgg cgccgtattc gctgagatcg gcccgagaat ggctgagctc
     2761 agcgggagta ggctcgcggc acagcagcaa atggagggac tgctggcacc acacaggccc
     2821 aaagaacccg cctggttcct ggcaaccgtc ggtgtatctc ccgatcatca ggggaaaggt
     2881 ctgggctctg ccgtagtgct ccctggcgtg gaggcagctg agagagcagg agtacctgcc
     2941 ttcttggaga cctccgctcc aaggaatctt cccttctatg aacggttggg cttcaccgtg
     3001 acagccgacg tggaagtccc cgaaggcccc cgcacttggt gcatgacgag gaagcctgga
     3061 gcgtgagggc ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     3121 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3181 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     3241 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     3301 taaaatcgat aaagatccgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
     3361 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
     3421 aactcgtagg acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc
     3481 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3541 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3601 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3661 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3721 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3781 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3841 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3901 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3961 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4021 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4081 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4141 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4201 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4261 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4321 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4381 tctgacagcg gccgcaaatg ctaaaccact gcagtggtta ccagtgcttg atcagtgagg
     4441 caccgatctc agcgatctgc ctatttcgtt cgtccatagt ggcctgactc cccgtcgtgt
     4501 agatcactac gattcgtgag ggcttaccat caggccccag cgcagcaatg atgccgcgag
     4561 agccgcgttc accggccccc gatttgtcag caatgaacca gccagcaggg agggccgagc
     4621 gaagaagtgg tcctgctact ttgtccgcct ccatccagtc tatgagctgc tgtcgtgatg
     4681 ctagagtaag aagttcgcca gtgagtagtt tccgaagagt tgtggccatt gctactggca
     4741 tcgtggtatc acgctcgtcg ttcggtatgg cttcgttcaa ctctggttcc cagcggtcaa
     4801 gccgggtcac atgatcaccc atattatgaa gaaatgcagt cagctcctta gggcctccga
     4861 tcgttgtcag aagtaagttg gccgcggtgt tgtcgctcat ggtaatggca gcactacaca
     4921 attctcttac cgtcatgcca tccgtaagat gcttttccgt gaccggcgag tactcaacca
     4981 agtcgttttg tgagtagtgt atacggcgac caagctgctc ttgcccggcg tctatacggg
     5041 acaacaccgc gccacatagc agtactttga aagtgctcat catcgggaat cgttcttcgg
     5101 ggcggaaaga ctcaaggatc ttgccgctat tgagatccag ttcgatatag cccactcttg
     5161 cacccagttg atcttcagca tcttttactt tcaccagcgt ttcggggtgt gcaaaaacag
     5221 gcaagcaaaa tgccgcaaag aagggaatga gtgcgacacg aaaatgttgg atgctcatac
     5281 tcgtcctttt tcaatattat tgaagcattt atcagggtta ctagtacgtc tctcaaggat
     5341 aagtaagtaa tattaaggta cgggaggtat tggacaggcc gc
//
