LOCUS       sequence-192511-        9376 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector encoding C-terminally FLAG tagged mouse PERK.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9376)
  AUTHORS   Taniuchi S, Miyake M, Tsugawa K, Oyadomari M, Oyadomari S
  TITLE     Integrated stress response of vertebrates is regulated by four 
            eIF2alpha kinases.
  JOURNAL   Sci Rep. 2016 Sep 16;6:32886. doi: 10.1038/srep32886.
  PUBMED    27633668
REFERENCE   2  (bases 1 to 9376)
  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: "Sci Rep."; 
            date: "2016-09-16"; volume: "6:32886. doi"; pages: " 
            10.1038/srep32886"
COMMENT     Sequence Label: pMXs-mPERK-FLAG-IP
FEATURES             Location/Qualifiers
     source          1..9376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             22..615
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    678..1035
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1094..1510
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1440..1462
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1478..1494
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1520..1894
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1832..1848
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1950..1959
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5331..5396
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    5545..6096
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5688..5705)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5915..5934
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6108..6710
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MVTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     6607..6627
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6881..7350
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(7480..7497)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7651..8239)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7731..7750)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8403..9194,9195..9263))
                     /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     9026..9045
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9264..9368)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta
       61 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc
      121 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      181 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
      241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
      301 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
      361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
      421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
      481 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
      541 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
      601 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
      661 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
      721 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
      781 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc
      841 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg
      901 actctttggt gcacccccct aataggaggg atatgtggtt ctggtaggag acgagaacct
      961 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc
     1021 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
     1081 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
     1141 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
     1201 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
     1261 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
     1321 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
     1381 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
     1441 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
     1501 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa
     1561 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca
     1621 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca
     1681 actggaccga ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg
     1741 acaccagact aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac
     1801 ccccaccgcc ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc
     1861 tgccgacccc gggggtggac catcctctag actgccggat ctagctagtt aattaaggat
     1921 cccagtgtgg tggtacggga attcctgcag ccaccatgga gcgcgccacc cggcccgggc
     1981 cgcgcgcgct gctgctgctt ctgttcctgc tgctgggctg cgcggcgggg atctcggcgg
     2041 tcgcgcccgc ccgcagtttg cttgctcccg cgtcggagac agtgtttggc ttaggggcag
     2101 cggccgcccc gacttcggcc gcgcgggtgc ctgcggtggc aacggccgaa gtgaccgtgg
     2161 aggacgccga ggcattgccg gctgccgctg gcgaaccgga gtcacgcgcg acggagcccg
     2221 atgacgacgt ggaactgcgg cctcgcggca ggtccttggt aatcatcagc actttagatg
     2281 gacgaatcgc tgcactggat gccgagaatg atgggaaaaa gcagtgggat ttggacgtgg
     2341 ggtctggttc cttggtttca tctagcctca gcaagccaga ggtgtttggg aacaagatga
     2401 tcatcccctc cctggatgga gacctcttcc agtgggaccg ggaccgagag agcatggagg
     2461 ccgtcccctt cacggtggag tccctgctcg aatcttccta caagtttgga gatgatgttg
     2521 ttctggttgg agggaaatct ctgactacat acggactcag tgcttacagt ggaaagctga
     2581 ggtatatctg ttctgccttg ggatgtcgcc gatgggatag tgatgaaatg gaagaagagg
     2641 aagacatctt gcttctgcag cgtacgcaga agactgtgcg agctgtcggg cctcgaagcg
     2701 gcagtgagaa gtggaatttc agtgttggcc actttgaact tcggtatatt ccagacatgg
     2761 aaactagagc cggattcatt gaaagcacct ttaaaccggg tggaaacaaa gaagactcta
     2821 aaattatttc agatgtggaa gaacaagaag ccaccatgct ggacacagtg ataaaagttt
     2881 ccgttgctga ttggaaggtc atggcgttta gtaggaaggg aggccgcctg gaatgggagt
     2941 accagttttg tactcccatc gcgtccgcct ggctggtgag ggatggcaag gtcatcccca
     3001 tcagcctgtt tgatgataca agttacacag ccagcgaaga agccttggga gacgaagaag
     3061 acattgtaga ggctgctcgg ggagccacag agaacagcgt gtacttaggg atgtacagag
     3121 gccagctgta cctgcagtcg tccgtcaggg tctcagaaaa gttccctaca agcccaaagg
     3181 ccttggagtc tgtaaatggc gaaaatgcaa ttattcctct gccgacgatc aaatggaagc
     3241 ccttaatcca ttctccttct aggactcctg tcttggttgg gtctgatgaa tttgacaaat
     3301 gtctaagtaa tgataagtat tcccacgaag aatacagtaa tggtgcactt tcaatcctcc
     3361 agtatccata cgataacggt tactatctgc catactacaa gagagaaagg aataagcgga
     3421 gcacgcagat cacagtcagg ttcctggaca gcccccacta cagcaagaac atccgcaaga
     3481 aggaccctat cctcctgctg cactggtgga aggagatatt cgggacgatc ctgctttgca
     3541 tcgtagccac gaccttcatc gtgcgcaggc ttttccatcc tcagccccac aggcagcgga
     3601 aggagtctga aactcagtgc cagactgaaa gtaaatacga ctccgtgagt gccgatgtca
     3661 gtgacaacag ctggaatgac atgaagtact caggatacgt atcccgatat ctaacagatt
     3721 ttgagccaat tcagtgcatg ggtcgtggtg gctttggcgt tgtctttgaa gctaaaaaca
     3781 aagtagatga ctgcaattac gctatcaaga ggatccggct ccccaacagg gagttggcac
     3841 gggagaaggt aatgcgggaa gttaaagcct tggctaagct ggaacaccca ggcattgtga
     3901 ggtatttcaa cgcctggctg gaaaccccac cagagaagtg gcaagaagag atggatgaga
     3961 tctggctcaa agacgaaagc acagactggc cgctcagctc ccctagcccg atggatgccc
     4021 catctgttaa gatccgaagg atggatcctt tctctacaaa agagcagatc gaagtcatag
     4081 ctccttctcc tgaaagaagt cggtctttct cggtgggcat ttcctgtggc cagacaagct
     4141 catcggagag ccagttctct cccctggagt tctcagggac agactgcgga gacaacagtg
     4201 actcagcgga cgcagcctac aacctccagg acagttgcct gacggactgc gaggacgtgg
     4261 aagatggcac cgtggacggc aatgacgagg gacactcctt tgaactttgt ccgtccgaag
     4321 cttctcccta tacccggtca agggaaggaa cgtcctcctc catagtgttt gaggactctg
     4381 gctgcggcaa cgcgtccagt aaggaggagc ccagagggaa ccggctgcat gatggcaacc
     4441 attatgttaa taagctaact gatctcaagt gctccagcag caggtcttct tcagaagcca
     4501 ccaccttgtc tacctcccct accaggccaa ccactctaag cttggatttc accaagaaca
     4561 ctgtgggcca gctccagccc agctccccca aggtgtatct gtacattcag atgcagctgt
     4621 gcaggaagga gaacctcaaa gactggatga accggcgctg cagcttggag gaccgggagc
     4681 acggcgtgtg cctgcacatc ttcctgcaga tcgcagaggc agtggagttc ctgcacagca
     4741 agggactcat gcacagggac ctcaagcctt ccaacatatt cttcacaatg gatgatgtgg
     4801 tcaaggttgg ggactttgga ctggtgactg ctatggacca agatgaagaa gagcagactg
     4861 tactgactcc aatgccagcc tatgctacgc acacgggaca agtagggacc aagctataca
     4921 tgagcccaga gcagattcat ggaaacaact actcccataa agtggacatc ttctctttag
     4981 gcttgattct gtttgaactc ctctacccat tcagcaccca gatggaacga gtccggattt
     5041 taactgatgt cagaaatctc aagtttcctc tactgttcac tcagaaatat ccccaagagc
     5101 atatgatggt tcaagacatg ctctctccat cccccacgga gcggcctgaa gccacagaca
     5161 tcattgaaaa tgccatattt gagaacttgg agtttcccgg gaaaacggtt ctgagacagc
     5221 ggtcccgctc catgagttca tctggaacaa aacattccag acagcccagc tgctcgtaca
     5281 gcccactgcc tggcaactgc aggcctcgac tcggtcgcga ggccaaggcc gactacaaag
     5341 accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt
     5401 gactgcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat
     5461 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttggccgcta cgtaaattcc
     5521 gccccccccc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
     5581 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttgcaatg tgagggcccg
     5641 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
     5701 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
     5761 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
     5821 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
     5881 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
     5941 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
     6001 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
     6061 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg gttaccgagt
     6121 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     6181 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     6241 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     6301 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     6361 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     6421 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     6481 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     6541 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     6601 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     6661 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gtcgacgata
     6721 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
     6781 tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac ataactgaga
     6841 atagagaagt tcagatcaag gtcaggaaca gatggaacag ctgaatatgg gccaaacagg
     6901 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa
     6961 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     7021 gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca
     7081 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
     7141 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
     7201 actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac
     7261 cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt
     7321 gattgactac ccgtcagcgg gggtctttca catgcagcat gtatcaaaat taatttggtt
     7381 ttttttctta agtatttaca ttaaatggcc atagttgcat taatgaatcg gccaacgcgc
     7441 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     7501 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     7561 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     7621 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     7681 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     7741 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7801 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7861 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7921 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7981 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     8041 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     8101 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     8161 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     8221 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     8281 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     8341 gatccttttg cggccggccg caaatcaatc taaagtatat atgagtaaac ttggtctgac
     8401 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     8461 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     8521 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     8581 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     8641 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     8701 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     8761 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     8821 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8881 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8941 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     9001 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     9061 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     9121 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     9181 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     9241 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     9301 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     9361 gttccgcgca catttc
//
