LOCUS       sequence-203459-        9102 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of ALDH7A1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9102)
  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 9102)
  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_ALDH7A1
FEATURES             Location/Qualifiers
     source          1..9102
                     /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    1683..1916
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2101..2145
                     /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    2443..2560
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2643..3022
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3023..3226
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3252..3271
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3252..3270
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4888..4917
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4936..4959
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    4993..5566
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5016..5568
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5160..5177)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5387..5406
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5568..6167
                     /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(5568..5587)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6064..6084
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(6197..6230)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6286..6874
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6339..6359)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6757..6768)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6877..6893)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6878..6894)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7403..7583
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7645..8233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7725..7744)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8310..8969)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     8884..8903
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(8970..9074)
                     /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 gctagaacga ttcgcagtta atcctggcct
     1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1501 aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat
     1561 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1621 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1681 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1741 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1801 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1861 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1921 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1981 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     2041 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     2101 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     2161 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     2221 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2281 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2341 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2401 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2461 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2521 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2581 cagagatcca gtttggttag taccgggccc gctctagaca tgtccaatat gaccgccatg
     2641 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     2701 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     2761 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     2821 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     2881 tcaagtgtat catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc
     2941 ctggcattat gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt
     3001 attagtcatc gctattacca tggtgatgcg gttttggcag tacaccaatg ggcgtggata
     3061 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     3121 ttggcaccaa aatcaacggg actttccaaa atgtcgtaat aaccccgccc cgttgacgca
     3181 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg
     3241 tcagaatttt gtaatacgac tcactatagg gcggccggga attcgtcgac tggatccggt
     3301 accgaggaga tctgccgccg cgatcgccgg cgcgccatgt ccactctcct catcaatcag
     3361 ccccagtatg cgtggctgaa agagctgggg ctccgcgagg aaaacgaggg cgtgtataat
     3421 ggaagctggg gaggccgggg agaggttatt acgacctatt gccctgctaa caacgagcca
     3481 atagcaagag tccgacaggc cagtgtggca gactatgaag aaactgtaaa gaaagcaaga
     3541 gaagcatgga aaatctgggc agatattcct gctccaaaac gaggagaaat agtaagacag
     3601 attggcgatg ccttgcggga gaagatccaa gtactaggaa gcttggtgtc tttggagatg
     3661 gggaaaatct tagtggaagg tgtgggtgaa gttcaggagt atgtggatat ctgtgactat
     3721 gctgttggtt tatcaaggat gattggagga cctatcttgc cttctgaaag atctggccat
     3781 gcactgattg agcagtggaa tcccgtaggc ctggttggaa tcatcacggc attcaatttc
     3841 cctgtggcag tgtatggttg gaacaacgcc atcgccatga tctgtggaaa tgtctgcctc
     3901 tggaaaggag ctccaaccac ttccctcatt agtgtggctg tcacaaagat aatagccaag
     3961 gttctggagg acaacaagct gcctggtgca atttgttcct tgacttgtgg tggagcagat
     4021 attggcacag caatggccaa agatgaacga gtgaacctgc tgtccttcac tgggagcact
     4081 caggtgggaa aacaggtggg cctgatggtg caggagaggt ttgggagaag tctgttggaa
     4141 cttggaggaa acaatgccat tattgccttt gaagatgcag acctcagctt agttgttcca
     4201 tcagctctct tcgctgctgt gggaacagct ggccagaggt gtaccactgc gaggcgactg
     4261 tttatacatg aaagcatcca tgatgaggtt gtaaacagac ttaaaaaggc ctatgcacag
     4321 atccgagttg ggaacccatg ggaccctaat gttctctatg ggccactcca caccaagcag
     4381 gcagtgagca tgtttcttgg agcagtggaa gaagcaaaga aagaaggtgg cacagtggtc
     4441 tatgggggca aggttatgga tcgccctgga aattatgtag aaccgacaat tgtgacaggt
     4501 cttggccacg atgcgtccat tgcacacaca gagacttttg ctccgattct ctatgtcttt
     4561 aaattcaaga atgaagaaga ggtctttgca tggaataatg aagtaaaaca gggactttca
     4621 agtagcatct ttaccaaaga tctgggcaga atctttcgct ggcttggacc taaaggatca
     4681 gactgtggca ttgtaaatgt caacattcca acaagtgggg ctgagattgg aggtgccttt
     4741 ggaggagaaa agcacactgg tggtggcagg gagtctggca gtgatgcctg gaaacagtac
     4801 atgagaaggt ctacttgtac tatcaactac agtaaagacc ttcctctggc ccaaggaatc
     4861 aagtttcaga cgcgtacgcg gccgctcgag cagaaactca tctcagaaga ggatctggca
     4921 gcaaatgata tcctggatta caaggatgac gacgataagg tttaaaccta ggcgtagcgg
     4981 ccgcaaattc cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga
     5041 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     5101 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     5161 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     5221 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     5281 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     5341 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     5401 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     5461 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     5521 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg accgagtaca
     5581 agcccacggt gcgcctcgcc acccgcgacg acgtcccccg ggcagtacgc accctcgccg
     5641 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccagaccgc cacatcgagc
     5701 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     5761 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     5821 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     5881 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     5941 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     6001 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     6061 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     6121 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgatgt acaagtagga
     6181 ttcgtcgagg gacctaataa cttcgtatag catacattat acgaagttat acatgtttaa
     6241 gggttccggt tccactaggt acaattcgat atcaagctta tcgataatca acctctggat
     6301 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     6361 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     6421 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     6481 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     6541 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     6601 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     6661 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     6721 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     6781 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     6841 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgatcgaga
     6901 cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc
     6961 ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt
     7021 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
     7081 actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca
     7141 cacacaaggc tacttccctg attggcagaa ctacacacca gggccaggga tcagatatcc
     7201 actgaccttt ggatggtgct acaagctagt accagttgag caagagaagg tagaagaagc
     7261 caatgaagga gagaacaccc gcttgttaca ccctgtgagc ctgcatggga tggatgaccc
     7321 ggagagagaa gtattagagt ggaggtttga cagccgccta gcatttcatc acatggcccg
     7381 agagctgcat ccggactgta ctgggtctct ctggttagac cagatctgag cctgggagct
     7441 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     7501 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     7561 gtcagtgtgg aaaatctcta gcagcatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     7621 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     7681 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7741 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7801 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7861 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7921 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7981 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     8041 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     8101 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     8161 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     8221 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     8281 aaactcacgt taagggattt tggtcatgat tacgccccgc cctgccactc atcgcagtac
     8341 tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc
     8401 tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa
     8461 acgggggcga agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc
     8521 cagggattgg ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg
     8581 ttttcaccgt aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg
     8641 tggtattcac tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa
     8701 gggtgaacac tatcccatat caccagctca ccgtctttca ttgccatacg gaactccgga
     8761 tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt
     8821 ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat
     8881 tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg
     8941 gtggtatatc cagtgatttt tttctccata ctcttccttt ttcaatatta ttgaagcatt
     9001 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     9061 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ac
//
