LOCUS       sequence-192509-        7795 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector encoding C-terminally FLAG tagged mouse HRI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7795)
  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 7795)
  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-mHRI-FLAG-IP
FEATURES             Location/Qualifiers
     source          1..7795
                     /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"
     CDS             3846..3911
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    3964..4515
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4107..4124)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4334..4353
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4527..5129
                     /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     5026..5046
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5300..5769
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5899..5916)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6070..6658)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6150..6169)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6822..7613,7614..7682))
                     /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     7445..7464
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7683..7787)
                     /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 ccaccatgct ggggggcagc tccgtggacg
     1981 gcgagcgcga cacggacgac gacgcggcgg gggcggtggc cgcgcctcct gccatcgact
     2041 tccccgcaga ggtgtcggac cccaagtatg atgagtccga tgtccccgca gagctccaag
     2101 tgttaaaaga gcccctacag cagcccacgt tccctttcct ggtggcaaac cagctgctgc
     2161 tcgtctcctt actggaacac ttgagccatg tgcacgagcc aaacccgctc cactccaaac
     2221 aggtgtttaa attactgtgc cagactttta tcaagatggg gctgctctct tcctttacct
     2281 gcagtgatga gtttagctct ctgagactcc accacaacag agccatcact catttaatga
     2341 ggtctgccaa agagagagtc cgtcaggatc cttgtcaaga taattcttac atgcagaaaa
     2401 tcagatccag agagatagcc ttcgaagcac aaacgtcacg ctacttaaat gaatttgaag
     2461 agcttgccat cttagggaaa ggaggatatg gaagagttta caaggtccgg aacaaattag
     2521 atggtcagca ttatgcaatt aagaaaatcc tgattaagag cgcaactaaa acagattgta
     2581 tgaaggtgct acgggaagtg aaggttctgg caggtctcca gcatcccaat attgttggct
     2641 accacactgc gtggatagaa catgttcatg tggttcagcc acaagacaga gttccgattc
     2701 aactgccctc tctcgaagtg ctgtcggagc aggaagggga cagagaccaa ggtggtgtta
     2761 aagataatga aagcagttcg tccattgtct ttgctgaact caccccagaa aaagaaaaac
     2821 cttttgggga gtctgaggtt aaaaatgaga ataacaacct ggtgagctac acggccaact
     2881 tagtggtcag gaacagcagt gaaagtgaat cgtccattga gctccaagaa gacggcttga
     2941 ctgatttgtc cgtcagacca gttgtcaggc atcagctgcc gctggggcat agctcggaat
     3001 tggaagggaa ttttacatcc acggatgagt cttctgaagg caacttgaac ctgctggggc
     3061 agacggaggt tcggtaccac ctgatgttgc acatccagat gcagctgtgt gagctctccc
     3121 tgtgggactg gataactgag aggaacaagc ggagccggga gtatgtggac gaagctgctt
     3181 gtccctatgt tatggccagt gttgcaacaa aaatttttca agaattggtg gaaggtgtct
     3241 tttacataca taacatgggc attgtccaca gagatctgaa gcctagaaat atttttcttc
     3301 atggccctga tcagcaagta aaaataggag actttggtct ggcctgtgca gacatcatcc
     3361 agaatgcaga ctggaccaac agaaatggga aaggaacacg gacacacaca tccagagtgg
     3421 ggacttgtct ctacgcatca ccggaacagc tggagggatc ccagtacgat gccaagtcag
     3481 atatgtatag cttgggtgtg atcctgctcg agctctttca gccattcggg acagaaatgg
     3541 agagggcaac agtcttaaca ggcgtaagga ctggtcggat acccgaatcc ctcagtaaaa
     3601 ggtgtccggt gcaagccaag tatatccagc tcctaaccgg gaggaatgtg tcacagagac
     3661 catctgccct tcagctgctg cagagtgagc tttttcaaac aactggaaat gttaatctca
     3721 cattgcagat gaagataata gaacaagaga aggaaattga agaactaaag aagcaactaa
     3781 gcctcctttc tcaggacaga gggctgaaga gatgcaggcc tcgactcggt cgcgaggcca
     3841 aggccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     3901 acgatgacaa gtagtgactg cggccgctac gtaaattccg cccccccccc tccccccccc
     3961 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     4021 tttccaccat attgccgtct tttgcaatgt gagggcccgg aaacctggcc ctgtcttctt
     4081 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     4141 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     4201 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     4261 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     4321 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     4381 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     4441 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     4501 aacacgatga taatatggcc acaaccatgg ttaccgagta caagcccacg gtgcgcctcg
     4561 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     4621 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     4681 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     4741 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     4801 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     4861 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     4921 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     4981 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     5041 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     5101 ggtgcatgac ccgcaagccc ggtgcctgag tcgacgataa aataaaagat tttatttagt
     5161 ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta
     5221 acgccatttt gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg
     5281 tcaggaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc
     5341 tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct
     5401 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     5461 agccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     5521 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     5581 tctgctcccc gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg
     5641 attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac
     5701 ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg
     5761 ggtctttcac atgcagcatg tatcaaaatt aatttggttt tttttcttaa gtatttacat
     5821 taaatggcca tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5881 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5941 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6001 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6061 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6121 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6181 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6241 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6301 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6361 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6421 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6481 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6541 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6601 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6661 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6721 gattttggtc atgagattat caaaaaggat cttcacctag atccttttgc ggccggccgc
     6781 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6841 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6901 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6961 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7021 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7081 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7141 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7201 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7261 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7321 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7381 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7441 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7501 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7561 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7621 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7681 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7741 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttc
//
