LOCUS       sequence-194347-        7506 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Retroviral vector encoding C-terminally FLAG tagged mouse PKR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7506)
  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 7506)
  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-mPKR-FLAG-IG
FEATURES             Location/Qualifiers
     source          1..7506
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    7..469
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(63..80)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     290..309
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(483..485,486..488,489..1202)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(483..485,486..488,489..1199)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(528..549)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(789..808)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1136..1157
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             1373..1842
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(1972..1989)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2143..2731)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2223..2242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2895..3686,3687..3755))
                     /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     3518..3537
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3756..3860)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             3890..4483
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    4546..4903
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             4962..5378
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5308..5330
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5346..5362
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    5388..5762
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     5700..5716
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      5818..5827
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7402..7467
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc
       61 ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga
      121 agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc
      181 tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc
      241 acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc
      301 aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga
      361 tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg
      421 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa
      481 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
      541 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      601 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
      661 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
      721 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
      781 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
      841 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
      901 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
      961 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     1021 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1081 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     1141 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     1201 aagtcgacga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     1261 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat
     1321 acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac agctgaatat
     1381 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     1441 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     1501 gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat
     1561 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
     1621 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
     1681 ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg
     1741 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg
     1801 tctcctctga gtgattgact acccgtcagc gggggtcttt cacatgcagc atgtatcaaa
     1861 attaatttgg ttttttttct taagtattta cattaaatgg ccatagttgc attaatgaat
     1921 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1981 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2041 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2101 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2161 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2221 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2281 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2341 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2401 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2461 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2521 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2581 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2641 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2701 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2761 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2821 gatcttcacc tagatccttt tgcggccggc cgcaaatcaa tctaaagtat atatgagtaa
     2881 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2941 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3001 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3061 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3121 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3181 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3241 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3301 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3361 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3421 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3481 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3541 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3601 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3661 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3721 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3781 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3841 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgcata atgaaagacc
     3901 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata
     3961 cataactgag aatagaaaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg
     4021 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     4081 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     4141 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc
     4201 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
     4261 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     4321 cacaacccct cactcggcgc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt
     4381 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
     4441 ctcctctgag tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga
     4501 tcgggagacc cctgcccagg gaccaccgac ccaccaccgg gaggtaagct ggccagcaac
     4561 ttatctgtgt ctgtccgatt gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta
     4621 ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg agttcggaac
     4681 acccggccgc aaccctggga gacgtcccag ggacttcggg ggccgttttt gtggcccgac
     4741 ctgagtccaa aaatcccgat cgttttggac tctttggtgc acccccctaa taggagggat
     4801 atgtggttct ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc
     4861 tttcggtttg ggaccgaagc cgcgccgcgc gtcttgtctg ctgcagcatc gttctgtgtt
     4921 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
     4981 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     5041 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     5101 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     5161 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     5221 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     5281 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     5341 cctcccttta tccagccctc actccttctc taggcgcccc catatggcca tatgagatct
     5401 tatatggggc acccccgccc cttgtaaact tccctgaccc tgacatgaca agagttacta
     5461 acagcccctc tctccaagct cacttacagg ctctctactt agtccagcac gaagtctgga
     5521 gacctctggc ggcagcctac caagaacaac tggaccgacc ggtggtacct cacccttacc
     5581 gagtcggcga cacagtgtgg gtccgccgac accagactaa gaacctagaa cctcgctgga
     5641 aaggacctta cacagtcctg ctgaccaccc ccaccgccct caaagtagac ggcatcgcag
     5701 cttggataca cgccgcccac gtgaaggctg ccgaccccgg gggtggacca tcctctagac
     5761 tgccggatct agctagttaa ttaaggatcc cagtgtggtg gtacgggaat tcctgcagcc
     5821 accatggcca gtgatacccc aggtttctac atggacaaac ttaataaata ccgccagatg
     5881 cacggagtag ccattacgta taaagaactt agtacttcgg gacctccaca tgacagaagg
     5941 tttacatttc aagttttaat agatgagaag gaatttccag aagccaaagg tagatcaaag
     6001 caggaggcaa gaaacgctgc agccaaatta gctgttgata tacttgataa cgaaaacaag
     6061 gtggattgtc acacgagtgc atctgagcaa ggcttgttcg ttggtaacta cataggcctt
     6121 gtcaatagct ttgcccagaa gaaaaagctg tctgtaaatt atgaacagtg tgagcccaac
     6181 tctgagttgc ctcaaagatt tatttgtaaa tgcaaaattg ggcagacaat gtatggtact
     6241 ggttcaggtg tcaccaaaca ggaggcaaag cagttggctg cgaaagaagc ctatcagaag
     6301 ctgttaaaga gcccgccgaa aactgccgga acatcctcta gcgttgtcac atctacattc
     6361 agtggctttt ccagcagctc gtctatgaca agtaatggtg tttcccagtc agcacctgga
     6421 agtttttcct cagagaacgt gtttacgaac ggtctcggag aaaataaaag gaaatcagga
     6481 gtaaaagtat cccctgatga tgtgcaaaga aataaatata ccttggacgc caggtttaac
     6541 agcgattttg aagacataga agaaattggc ttaggtggat ttggtcaagt tttcaaagcg
     6601 aaacacagaa ttgatggaaa gagatacgct attaagcgcg ttaaatataa cacggagaag
     6661 gcggagcacg aagtacaagc gctggcagaa ctcaatcacg tcaacattgt ccaataccat
     6721 agttgttggg agggagttga ctatgatcct gagcacagca tgagtgatac aagtcgatac
     6781 aaaacccggt gcctctttat tcaaatggaa ttctgtgata aaggaacttt ggagcaatgg
     6841 atgagaaaca gaaatcagag taaagtggac aaagctttga ttttggactt atatgaacaa
     6901 atcgtgaccg gagtggagta tatacactcg aaagggttaa ttcacagaga tcttaagcca
     6961 ggtaatatat ttttagtaga tgaaagacac attaagatcg gagactttgg ccttgcaaca
     7021 gccctggaaa atgatggaaa atcccgaaca aggagaacag gaactcttca atacatgagt
     7081 ccagaacagt tatttttaaa gcactatgga aaagaagtgg acatctttgc tttgggcctt
     7141 attctagctg aacttcttca cacgtgcttc acggagtcag agaaaataaa gtttttcgaa
     7201 agtctaagaa aaggcgactt ctctaatgat atattcgaca acaaagaaaa aagccttcta
     7261 aaaaaactac tctcagagaa acccaaggac cgacctgaga catctgaaat cctgaagacc
     7321 ttggctgaat ggaggaacat ctcagagaaa aagaaaagaa acacatgttg caggcctcga
     7381 ctcggtcgcg aggccaaggc cgactacaaa gaccatgacg gtgattataa agatcatgac
     7441 atcgactaca aggatgacga tgacaagtag tgactgcggc cgctacgtaa attccgcccc
     7501 cccccc
//
