LOCUS       sequence-234791-        7893 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of PRKRA delta 12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7893)
  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 7893)
  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_delta12
FEATURES             Location/Qualifiers
     source          1..7893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        94..473
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        474..677
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     703..722
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        703..721
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1130..1159
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1178..1201
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1235..1808
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1258..1810
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1402..1419)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1629..1648
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1810..2409
                     /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(1810..1829)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2306..2326
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(2439..2472)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2528..3116
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2581..2601)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2999..3010)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3119..3135)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3120..3136)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3645..3825
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3887..4475)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3967..3986)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4552..5211)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5126..5145
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5212..5316)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5391..5410)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5581..5960
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5962..6160
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6178..6358
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6405..6530
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7027..7260
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7445..7489
                     /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    join(7787..7893,1..11)
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ttcaaaattt tcgggtttat tacagggaca gcagagatcc agtttggtta gtaccgggcc
       61 cgctctagac atgtccaata tgaccgccat gttgacattg attattgact agttattaat
      121 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      181 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      241 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      301 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc
      361 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac
      421 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      481 ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      541 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      601 aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg
      661 tctatataag cagagctcgt ttagtgaacc gtcagaattt tgtaatacga ctcactatag
      721 ggcggccggg aattcgtcga ctggatccat gaatatttct ccagagaacc acatttcttt
      781 aacaaatgta gtaggacatt ctttaggatg tacttggcat tccttgagga attctcctgg
      841 tgaaaagatc aacttactga aaagaagcct ccttagtatt ccaaatacag attacatcca
      901 gctgcttagt gaaattgcca aggaacaagg ttttaatata acatatttgg atatagatga
      961 actgagcgcc aatggacaat atcaatgtct tgctgaactg tccaccagcc ccatcacagt
     1021 ctgtcatggc tccggtatct cctgtggcaa tgcacaaagt gatgcagctc acaatgcttt
     1081 gcagtattta aagataatag cagaaagaaa gacgcgtacg cggccgctcg agcagaaact
     1141 catctcagaa gaggatctgg cagcaaatga tatcctggat tacaaggatg acgacgataa
     1201 ggtttaaacc taggcgtagc ggccgcaaat tccgcccctc tccctccccc ccccctaacg
     1261 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     1321 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     1381 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     1441 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     1501 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     1561 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     1621 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     1681 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     1741 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     1801 gatgataata tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     1861 cgggcagtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     1921 gatccagacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     1981 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     2041 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     2101 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     2161 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     2221 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     2281 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     2341 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     2401 ggtgcctgat gtacaagtag gattcgtcga gggacctaat aacttcgtat agcatacatt
     2461 atacgaagtt atacatgttt aagggttccg gttccactag gtacaattcg atatcaagct
     2521 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2581 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2641 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2701 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     2761 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     2821 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2881 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     2941 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3001 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3061 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     3121 ataccgtcga cctcgatcga gacctagaaa aacatggagc aatcacaagt agcaatacag
     3181 cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc
     3241 cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     3301 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata
     3361 tccttgatct gtggatctac cacacacaag gctacttccc tgattggcag aactacacac
     3421 cagggccagg gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg
     3481 agcaagagaa ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga
     3541 gcctgcatgg gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc
     3601 tagcatttca tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag
     3661 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     3721 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     3781 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagcatg tgagcaaaag
     3841 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3901 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3961 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4021 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4081 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4141 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4201 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4261 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4321 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4381 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4441 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4501 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg attacgcccc
     4561 gccctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc
     4621 atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt
     4681 ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa
     4741 atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa
     4801 ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg
     4861 tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg
     4921 ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt
     4981 cattgccata cggaactccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc
     5041 cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg
     5101 aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg
     5161 atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca tactcttcct
     5221 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5281 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5341 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     5401 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     5461 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     5521 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatcgcgtt
     5581 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     5641 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     5701 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     5761 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     5821 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     5881 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     5941 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     6001 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     6061 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     6121 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
     6181 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     6241 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     6301 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     6361 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
     6421 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
     6481 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
     6541 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     6601 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     6661 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     6721 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     6781 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     6841 acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac ctggaggagg
     6901 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     6961 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     7021 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     7081 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     7141 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     7201 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     7261 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     7321 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     7381 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     7441 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     7501 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     7561 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     7621 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     7681 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc
     7741 actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag
     7801 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     7861 aaactaaaga attacaaaaa caaattacaa aaa
//
