LOCUS       sequence-424952-        6204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA tagged human ERK2 R67S D321N from puromycin resistance 
            retroviral vector .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6204)
  AUTHORS   Brady DC, Crowe MS, Turski ML, Hobbs GA, Yao X, Chaikuad A, Knapp S,
            Xiao K, Campbell SL, Thiele DJ, Counter CM
  TITLE     Copper is required for oncogenic BRAF signalling and tumorigenesis.
  JOURNAL   Nature. 2014 Apr 9. doi: 10.1038/nature13180.
  PUBMED    24717435
REFERENCE   2  (bases 1 to 6204)
  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: "Nature. 
            2014 Apr 9. doi: 10.1038/nature13180."
COMMENT     Sequence Label: pBABEpuro-HA-ERK2 GOF
FEATURES             Location/Qualifiers
     source          1..6204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             347..763
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     693..715
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     731..747
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             818..844
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1889..1924
                     /codon_start=1
                     /product="epitope tag derived from MAP kinase 2"
                     /label=Tag-100
                     /translation="EETARFQPGYRS"
     primer_bind     complement(1939..1959)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1944..2273
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2124..2259
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2283..2882
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2283..2302)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2779..2799
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3103..3572
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3702..3719)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3873..4461)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3953..3972)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4621..5412,5413..5481))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5244..5263
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5482..5586)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5617..6086
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    join(6151..6204,1..146)
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 ttttatgcgc ctgcgtcggt actagttagc taactagctc tgtatctggc ggacccgtgg
       61 tggaactgac gagttctgaa cacccggccg caaccctggg agacgtccca gggactttgg
      121 gggccgtttt tgtggcccga cctgaggaag ggagtcgatg tggaatccga ccccgtcagg
      181 atatgtggtt ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt
      241 gctttcggtt tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct
      301 gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc agactgttac
      361 cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg ctcacaacca
      421 gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat ggccaacctt
      481 taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc aggttaagat
      541 caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca tcgtgacctg
      601 ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc ctaagcctcc
      661 gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc
      721 tcgatcctcc ctttatccag ccctcactcc ttctctaggc gccggccgga tcccacgcgt
      781 taactcgcga gctctcgagg atatcgccgc caccatgtac ccatacgatg ttccagatta
      841 cgctgcggcg gcggcggcgg cgggcgcggg cccggagatg gtccgcgggc aggtgttcga
      901 cgtggggccg cgctacacca acctctcgta catcggcgag ggcgcctacg gcatggtgtg
      961 ctctgcttat gataatgtca acaaagttcg agtagctatc aagaaaatca gcccctttga
     1021 gcaccagacc tactgccaga gtaccctgag ggagataaaa atcttactgc gcttcagaca
     1081 tgagaacatc attggaatca atgacattat tcgagcacca accatcgagc aaatgaaaga
     1141 tgtatatata gtacaggacc tcatggaaac agatctttac aagctcttga agacacaaca
     1201 cctcagcaat gaccatatct gctattttct ctaccagatc ctcagagggt taaaatatat
     1261 ccattcagct aacgttctgc accgtgacct caagccttcc aacctgctgc tcaacaccac
     1321 ctgtgatctc aagatctgtg actttggcct ggcccgtgtt gcagatccag accatgatca
     1381 cacagggttc ctgacagaat atgtggccac acgttggtac agggctccag aaattatgtt
     1441 gaattccaag ggctacacca agtccattga tatttggtct gtaggctgca ttctggcaga
     1501 aatgctttct aacaggccca tctttccagg gaagcattat cttgaccagc tgaaccacat
     1561 tttgggtatt cttggatccc catcacaaga agacctgaat tgtataataa atttaaaagc
     1621 taggaactat ttgctttctc ttccacacaa aaataaggtg ccatggaaca ggctgttccc
     1681 aaatgctgac tccaaagctc tggacttatt ggacaaaatg ttgacattca acccacacaa
     1741 gaggattgaa gtagaacagg ctctggccca cccatatctg gagcagtatt acgacccgag
     1801 tagtaacgag cccatcgccg aagcaccatt caagttcgac atggaattgg atgacttgcc
     1861 taaggaaaag cccaaagaac taatttttga agagactgct agattccagc caggatacag
     1921 atcttaagtc gaccctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     1981 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     2041 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2101 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     2161 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     2221 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta
     2281 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     2341 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     2401 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     2461 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     2521 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     2581 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     2641 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     2701 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     2761 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     2821 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     2881 gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcga
     2941 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     3001 ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga
     3061 gaatagagaa gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca
     3121 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg
     3181 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     3241 cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc
     3301 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     3361 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     3421 tcactcgggg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa
     3481 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     3541 gtgattgact acccgtcagc gggggtcttt cacatgcagc atgtatcaaa attaatttgg
     3601 ttttttttct taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc
     3661 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3721 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3781 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3841 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3901 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3961 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4021 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4081 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4141 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4201 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4261 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4321 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4381 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4441 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4501 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4561 tagatccttt tgcggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4621 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4681 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4741 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4801 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4861 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4921 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4981 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5041 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5101 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5161 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5221 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5281 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5341 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5401 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5461 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5521 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5581 tccgcgcaca tttccccgaa aagtgccacc tgcagcctga atatgggcca aacaggatat
     5641 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg
     5701 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     5761 gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt
     5821 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
     5881 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc
     5941 ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc
     6001 ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt
     6061 gactacccgt cagcgggggt ctttcatttg ggggctcgtc cgggatcggg agacccctgc
     6121 ccagggacca ccgacccacc accgggaggc aagctggcca gcaacttatc tgtgtctgtc
     6181 cgattgtcta gtgtctatga ctga
//
