LOCUS       sequence-228369-        8286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of PRKRA delta3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8286)
  AUTHORS   Meyer C, Garzia A, Mazzola M, Gerstberger S, Molina H, Tuschl T
  TITLE     The TIA1 RNA-Binding Protein Family Regulates EIF2AK2-Mediated 
            Stress Response and Cell Cycle Progression.
  JOURNAL   Mol Cell. 2018 Feb 15;69(4):622-635.e6. doi: 
            10.1016/j.molcel.2018.01.011.
  PUBMED    29429924
REFERENCE   2  (bases 1 to 8286)
  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"; doi: 
            "10.1016/j.molcel.2018.01"; journalName: "Mol Cell"; date: 
            "2018-02-15- 15"; volume: "69"; issue: "4"; pages: "622-635.e6"
COMMENT     Sequence Label: pLenti_C_Myc_DDK_IRES_Puro_PRKRA_delta3
FEATURES             Location/Qualifiers
     source          1..8286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1447..1680
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1865..1909
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2207..2324
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2407..2786
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2787..2990
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3016..3035
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3016..3034
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3836..3865
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3884..3907
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    3941..4514
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3964..4516
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4108..4125)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4335..4354
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4516..5115
                     /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(4516..4535)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5012..5032
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(5145..5178)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5234..5822
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5287..5307)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5705..5716)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5825..5841)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5826..5842)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6351..6531
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6593..7181)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6673..6692)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7258..7917)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7832..7851
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7918..8022)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8097..8116)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac ctggaggagg
     1321 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     1381 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     1441 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     1501 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     1561 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     1621 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     1681 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     1741 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     1801 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     1861 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     1921 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     1981 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     2041 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     2101 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc
     2161 actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag
     2221 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2281 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2341 acagcagaga tccagtttgg ttagtaccgg gcccgctcta gacatgtcca atatgaccgc
     2401 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     2461 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     2521 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     2581 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     2641 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
     2701 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
     2761 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
     2821 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     2881 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
     2941 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
     3001 accgtcagaa ttttgtaata cgactcacta tagggcggcc gggaattcgt cgactggatc
     3061 cggtaccgag gagatctgcc gccgcgatcg ccggcgcgcc atgtcccaga gcaggcaccg
     3121 cgccgaggcc ccgccgctgg agcgcgagga cagtgggacc ttcagtttgg ggaagatgat
     3181 aacagctaag ccagggaaaa caccgattca ggtattacac gaatacggca tgaagaccaa
     3241 gaacatccca gtttatgaat gtgaaagatc tgatgtgcaa atacacgtgc ccactttcac
     3301 cttcagagta accgttggtg acataacctg cacaggtgaa ggtacaagta agaagctggc
     3361 gaaacataga gctgcagagg ctgccataaa cattttgaaa gccaatgcaa gtatttgctt
     3421 tgcagttcct gaccccttaa tgcctgaccc ttccaagcaa ccaaagaacc agcttaatcc
     3481 tattggttca ttacaggaat tggctattca tcatggctgg agacttcctg aatataccct
     3541 ttcccaggag ggaggacctg ctcataagag agaatatact acaatttgca ggctagagtc
     3601 atttatggaa actggaaagg gggcatcaaa aaagcaagcc aaaaggaatg ctgctgagaa
     3661 atttcttgcc aaatttagta atatttctcc agagaaccac atttctttaa caaatgtagt
     3721 aggacattct ttaggatgta cttggcattc cttgaggaat tctcctggtg aaaagatcaa
     3781 cttactgaaa agaagcctcc ttagtattcc aaatacaacg cgtacgcggc cgctcgagca
     3841 gaaactcatc tcagaagagg atctggcagc aaatgatatc ctggattaca aggatgacga
     3901 cgataaggtt taaacctagg cgtagcggcc gcaaattccg cccctctccc tccccccccc
     3961 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     4021 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     4081 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     4141 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc
     4201 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     4261 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     4321 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     4381 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt
     4441 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     4501 aaacacgatg ataatatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     4561 gtcccccggg cagtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     4621 accgtcgatc cagaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     4681 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     4741 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     4801 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     4861 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     4921 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     4981 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     5041 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     5101 aagcccggtg cctgatgtac aagtaggatt cgtcgaggga cctaataact tcgtatagca
     5161 tacattatac gaagttatac atgtttaagg gttccggttc cactaggtac aattcgatat
     5221 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     5281 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     5341 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     5401 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     5461 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     5521 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     5581 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     5641 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     5701 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     5761 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     5821 gcatcgatac cgtcgacctc gatcgagacc tagaaaaaca tggagcaatc acaagtagca
     5881 atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg
     5941 gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     6001 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
     6061 aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat tggcagaact
     6121 acacaccagg gccagggatc agatatccac tgacctttgg atggtgctac aagctagtac
     6181 cagttgagca agagaaggta gaagaagcca atgaaggaga gaacacccgc ttgttacacc
     6241 ctgtgagcct gcatgggatg gatgacccgg agagagaagt attagagtgg aggtttgaca
     6301 gccgcctagc atttcatcac atggcccgag agctgcatcc ggactgtact gggtctctct
     6361 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     6421 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     6481 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agcatgtgag
     6541 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6601 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6661 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6721 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6781 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6841 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6901 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6961 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7021 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7081 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7141 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7201 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgatta
     7261 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
     7321 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     7381 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
     7441 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc
     7501 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
     7561 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
     7621 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
     7681 gtctttcatt gccatacgga actccggatg agcattcatc aggcgggcaa gaatgtgaat
     7741 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
     7801 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc
     7861 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatact
     7921 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7981 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8041 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     8101 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     8161 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     8221 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     8281 cgcgtt
//
