LOCUS       sequence-203451-        7946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of PRKRA delta23.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7946)
  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 7946)
  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_delta23
FEATURES             Location/Qualifiers
     source          1..7946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..816
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             834..1014
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1061..1186
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1679..1912
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2097..2141
                     /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    2439..2556
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2639..3018
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3019..3222
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3248..3267
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3248..3266
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3732..3761
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3780..3803
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    3837..4410
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3860..4412
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4004..4021)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4231..4250
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4412..5011
                     /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(4412..4431)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4908..4928
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(5041..5074)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5130..5718
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5183..5203)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5601..5612)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5721..5737)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5722..5738)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6247..6427
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6489..7077)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6569..6588)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7154..7813)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7728..7747
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7814..7918)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
      841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     1261 attaaaacat atagtatggg caagcaggga gctcgattcg cagttaatcc tggcctgtta
     1321 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
     1381 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
     1441 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
     1501 aagaccaccg cacagcaagc ggccggccgc tgatcttcag acctggagga ggagatatga
     1561 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag
     1621 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag
     1681 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga
     1741 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc
     1801 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc
     1861 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt
     1921 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta
     1981 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta
     2041 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga
     2101 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa
     2161 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa
     2221 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat
     2281 cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag
     2341 aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg
     2401 cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt
     2461 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     2521 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
     2581 gatccagttt ggttagtacc gggcccgctc tagacatgtc caatatgacc gccatgttga
     2641 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     2701 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     2761 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     2821 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     2881 gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     2941 cattatgccc agtacatgac cttacgggac tttcctactt ggcagtacat ctacgtatta
     3001 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg tggatagcgg
     3061 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     3121 caccaaaatc aacgggactt tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg
     3181 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag
     3241 aattttgtaa tacgactcac tatagggcgg ccgggaattc gtcgactgga tccggtaccg
     3301 aggagatctg ccgccgcgat cgccggcgcg ccatgtccca gagcaggcac cgcgccgagg
     3361 ccccgccgct ggagcgcgag gacagtggga ccttcagttt ggggaagatg ataacagcta
     3421 agccagggaa aacaccgatt caggtattac acgaatacgg catgaagacc aagaacatcc
     3481 cagtttatga atgtgaaaga tctgatgtgc aaatacacgt gcccactttc accttcagag
     3541 taaccgttgg tgacataacc tgcacaggtg aaggtacaag taagaagctg gcgaaacata
     3601 gagctgcaga ggctgccata aacattttga aagccaatgc aagtatttgc tttgcagttc
     3661 ctgacccctt aatgcctgac ccttccaagc aaccaaagaa ccagcttaat cctacgcgta
     3721 cgcggccgct cgagcagaaa ctcatctcag aagaggatct ggcagcaaat gatatcctgg
     3781 attacaagga tgacgacgat aaggtttaaa cctaggcgta gcggccgcaa attccgcccc
     3841 tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     3901 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     3961 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     4021 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     4081 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     4141 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     4201 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     4261 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     4321 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     4381 gtggttttcc tttgaaaaac acgatgataa tatgaccgag tacaagccca cggtgcgcct
     4441 cgccacccgc gacgacgtcc cccgggcagt acgcaccctc gccgccgcgt tcgccgacta
     4501 ccccgccacg cgccacaccg tcgatccaga ccgccacatc gagcgggtca ccgagctgca
     4561 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     4621 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     4681 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     4741 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     4801 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     4861 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     4921 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     4981 ctggtgcatg acccgcaagc ccggtgcctg atgtacaagt aggattcgtc gagggaccta
     5041 ataacttcgt atagcataca ttatacgaag ttatacatgt ttaagggttc cggttccact
     5101 aggtacaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     5161 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     5221 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     5281 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     5341 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     5401 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     5461 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     5521 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     5581 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     5641 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     5701 tttgggccgc ctccccgcat cgataccgtc gacctcgatc gagacctaga aaaacatgga
     5761 gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa
     5821 gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac
     5881 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     5941 cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc
     6001 cctgattggc agaactacac accagggcca gggatcagat atccactgac ctttggatgg
     6061 tgctacaagc tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac
     6121 acccgcttgt tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta
     6181 gagtggaggt ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggac
     6241 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     6301 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     6361 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     6421 tctagcagca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6481 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6541 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6601 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6661 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6721 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6781 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6841 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6901 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6961 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7021 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7081 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7141 attttggtca tgattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa
     7201 gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca
     7261 tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt
     7321 tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga
     7381 cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg
     7441 ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga
     7501 gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc
     7561 atatcaccag ctcaccgtct ttcattgcca tacggaactc cggatgagca ttcatcaggc
     7621 gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta
     7681 aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa
     7741 atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga
     7801 tttttttctc catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7861 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7921 catttccccg aaaagtgcca cctgac
//
