LOCUS       sequence-192510-        7600 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector encoding C-terminally FLAG tagged mouse PKR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7600)
  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 7600)
  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..7600
                     /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             3534..3599
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    3652..4203
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3795..3812)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4022..4041
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4215..4217,4218..4220,4221..4934)
                     /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(4215..4217,4218..4220,4221..4931)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4260..4281)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4521..4540)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4868..4889
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5105..5574
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5704..5721)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5875..6463)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5955..5974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6627..7418,7419..7487))
                     /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     7250..7269
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7488..7592)
                     /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 ccaccatggc cagtgatacc ccaggtttct
     1981 acatggacaa acttaataaa taccgccaga tgcacggagt agccattacg tataaagaac
     2041 ttagtacttc gggacctcca catgacagaa ggtttacatt tcaagtttta atagatgaga
     2101 aggaatttcc agaagccaaa ggtagatcaa agcaggaggc aagaaacgct gcagccaaat
     2161 tagctgttga tatacttgat aacgaaaaca aggtggattg tcacacgagt gcatctgagc
     2221 aaggcttgtt cgttggtaac tacataggcc ttgtcaatag ctttgcccag aagaaaaagc
     2281 tgtctgtaaa ttatgaacag tgtgagccca actctgagtt gcctcaaaga tttatttgta
     2341 aatgcaaaat tgggcagaca atgtatggta ctggttcagg tgtcaccaaa caggaggcaa
     2401 agcagttggc tgcgaaagaa gcctatcaga agctgttaaa gagcccgccg aaaactgccg
     2461 gaacatcctc tagcgttgtc acatctacat tcagtggctt ttccagcagc tcgtctatga
     2521 caagtaatgg tgtttcccag tcagcacctg gaagtttttc ctcagagaac gtgtttacga
     2581 acggtctcgg agaaaataaa aggaaatcag gagtaaaagt atcccctgat gatgtgcaaa
     2641 gaaataaata taccttggac gccaggttta acagcgattt tgaagacata gaagaaattg
     2701 gcttaggtgg atttggtcaa gttttcaaag cgaaacacag aattgatgga aagagatacg
     2761 ctattaagcg cgttaaatat aacacggaga aggcggagca cgaagtacaa gcgctggcag
     2821 aactcaatca cgtcaacatt gtccaatacc atagttgttg ggagggagtt gactatgatc
     2881 ctgagcacag catgagtgat acaagtcgat acaaaacccg gtgcctcttt attcaaatgg
     2941 aattctgtga taaaggaact ttggagcaat ggatgagaaa cagaaatcag agtaaagtgg
     3001 acaaagcttt gattttggac ttatatgaac aaatcgtgac cggagtggag tatatacact
     3061 cgaaagggtt aattcacaga gatcttaagc caggtaatat atttttagta gatgaaagac
     3121 acattaagat cggagacttt ggccttgcaa cagccctgga aaatgatgga aaatcccgaa
     3181 caaggagaac aggaactctt caatacatga gtccagaaca gttattttta aagcactatg
     3241 gaaaagaagt ggacatcttt gctttgggcc ttattctagc tgaacttctt cacacgtgct
     3301 tcacggagtc agagaaaata aagtttttcg aaagtctaag aaaaggcgac ttctctaatg
     3361 atatattcga caacaaagaa aaaagccttc taaaaaaact actctcagag aaacccaagg
     3421 accgacctga gacatctgaa atcctgaaga ccttggctga atggaggaac atctcagaga
     3481 aaaagaaaag aaacacatgt tgcaggcctc gactcggtcg cgaggccaag gccgactaca
     3541 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     3601 agtgactgcg gccgctacgt aaattccgcc cccccccctc ccccccccct aacgttactg
     3661 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
     3721 tgccgtcttt tgcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
     3781 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
     3841 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
     3901 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
     3961 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
     4021 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
     4081 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa
     4141 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata
     4201 atatggccac aaccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     4261 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     4321 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     4381 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     4441 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     4501 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     4561 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     4621 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     4681 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     4741 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     4801 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     4861 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     4921 agctgtacaa gtaagtcgac gataaaataa aagattttat ttagtctcca gaaaaagggg
     4981 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     5041 gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg aacagatgga
     5101 acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     5161 ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc
     5221 tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc tcagcagttt
     5281 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     5341 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     5401 caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc
     5461 cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt
     5521 tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcacatgca
     5581 gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagtt
     5641 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5701 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5761 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5821 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5881 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5941 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6001 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6061 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6121 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6181 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6241 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6301 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6361 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6421 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6481 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6541 attatcaaaa aggatcttca cctagatcct tttgcggccg gccgcaaatc aatctaaagt
     6601 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6661 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6721 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6781 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6841 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6901 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6961 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7021 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7081 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7141 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7201 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7261 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7321 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7381 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7441 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7501 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7561 atttagaaaa ataaacaaat aggggttccg cgcacatttc
//
