LOCUS       sequence-448395-        9141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of protein in mammalian cell culture, rescue 
            experiments.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9141)
  AUTHORS   Adriaenssens E, Schaar S, Cook ASI, Stuke JFM, Sawa-Makarska J, 
            Nguyen TN, Ren X, Schuschnig M, Romanov J, Khuu G, Uoselis L, 
            Lazarou M, Hummer G, Hurley JH, Martens S
  TITLE     Reconstitution of BNIP3/NIX-mitophagy initiation reveals 
            hierarchical flexibility of the autophagy machinery.
  JOURNAL   Nat Cell Biol. 2025 Jul 25. doi: 10.1038/s41556-025-01712-y.
  PUBMED    40715440
REFERENCE   2  (bases 1 to 9141)
  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: "Nat Cell 
            Biol. 2025 Jul 25. doi: 10.1038/s41556-025-01712-y."
COMMENT     Sequence Label: pGenlenti_ATG13-delta191-205
FEATURES             Location/Qualifiers
     source          1..9141
                     /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             4837..4860
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    4894..5467
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4917..5469
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5061..5078)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5288..5307
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5469..6068
                     /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(5469..5488)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5965..5985
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6124..6712
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6177..6197)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6595..6606)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6715..6731)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6716..6732)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7241..7421
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7483..8071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7563..7582)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8148..8939,8940..9008))
                     /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     8771..8790
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9009..9113)
                     /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 tggatccatg
     3301 gaaactgatc tcaattccca ggacagaaag gacctggaca agtttattaa attttttgcc
     3361 ctcaagactg tccaagtgat tgtccaggct cggcttggtg aaaagatttg cactcgttca
     3421 tcatcttctc caacgggttc agattggttc aacttagcaa tcaaagacat cccagaggtt
     3481 acacatgaag caaagaaggc actggcagga cagctgcctg cagtcgggag gtccatgtgt
     3541 gtggagattt cacttaagac ttctgaggga gattccatgg agctggaaat atggtgtctt
     3601 gaaatgaatg aaaagtgtga taaagaaatc aaagtttcct acacggtgta caacagactg
     3661 tcattgctgc tgaagtccct tcttgctata actagggtga caccagccta taggctctcc
     3721 aggaaacaag ggcatgaata tgtcatatta tacaggatat attttggaga agttcagctg
     3781 agtggcttag gagaaggctt ccagacagtt cgtgttggga cagtgggcac ccctgtgggc
     3841 accatcactc tttcttgtgc ttacagacct atcatgggga ttattattga tcactttgtg
     3901 gaccgtccct atcccagctc ctctcccatg cacccctgca attacagaac tgctggtgag
     3961 gacactggag taatataccc gtctgtagaa gactctcaag aagtgtgtac cacctctttt
     4021 tccacctccc caccatccca gctctcaagc tctcgccttt cctatcagcc tgctgccctg
     4081 ggcgttggat cagctgacct ggcttatcca gtagtgtttg ctgctggctt aaatgctaca
     4141 caccctcacc agctgatggt tcctgggaag gaaggtgggg taccccttgc tcccaaccag
     4201 cctgtccatg gtacccaggc tgaccaggag agactggcaa cctgcacccc ttctgacaga
     4261 acccactgtg ctgccacacc ctccagtagt gaggatactg aaaccgtatc aaacagcagt
     4321 gagggacggg cctcccctca cgatgtcttg gagaccatct ttgtccgaaa agtgggggct
     4381 tttgtcaaca aacccattaa ccaggtgacc ctgacgagtt tggatatacc ctttgccatg
     4441 tttgctccca agaatttgga gctggaggat accgatccaa tggtgaatcc tccagattcc
     4501 ccagagactg aatctcctct ccagggcagc ctgcactcag atggctccag cgggggcagc
     4561 agtggcaata cccatgatga ctttgttatg atagacttta aaccagcttt ttctaaagat
     4621 gacattcttc cgatggacct ggggaccttc tatcgggagt ttcagaaccc acctcagctg
     4681 agcagcctct ccatagatat tggagcacag tccatggctg aagacttgga ctcattacca
     4741 gagaagctgg ctgtgcatga gaagaatgtc cgcgagtttg atgcctttgt ggaaaccctg
     4801 cagtaagcta gcggaccgac gcgtacgcgg ccgctcgatt acaaggatga cgacgataag
     4861 gtttaaacct aggcgtagcg gccgcaaatt ccgcccctct ccctcccccc cccctaacgt
     4921 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
     4981 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
     5041 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
     5101 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
     5161 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     5221 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     5281 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
     5341 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag
     5401 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     5461 atgataatat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc
     5521 gggcagtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
     5581 atccagaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
     5641 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
     5701 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
     5761 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
     5821 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
     5881 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
     5941 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
     6001 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
     6061 gtgcctgaac atgtttaagg gttccggttc cactaggtac aattcgatat caagcttatc
     6121 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     6181 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     6241 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     6301 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     6361 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     6421 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     6481 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     6541 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     6601 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     6661 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     6721 cgtcgacctc gatcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc
     6781 taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt
     6841 cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     6901 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatatcct
     6961 tgatctgtgg atctaccaca cacaaggcta cttccctgat tggcagaact acacaccagg
     7021 gccagggatc agatatccac tgacctttgg atggtgctac aagctagtac cagttgagca
     7081 agagaaggta gaagaagcca atgaaggaga gaacacccgc ttgttacacc ctgtgagcct
     7141 gcatgggatg gatgacccgg agagagaagt attagagtgg aggtttgaca gccgcctagc
     7201 atttcatcac atggcccgag agctgcatcc ggactgtact gggtctctct ggttagacca
     7261 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     7321 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     7381 atccctcaga cccttttagt cagtgtggaa aatctctagc agcatgtgag caaaaggcca
     7441 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7501 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7561 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7621 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7681 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7741 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7801 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7861 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7921 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7981 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     8041 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     8101 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgatta ccaatgctta
     8161 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     8221 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     8281 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     8341 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     8401 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     8461 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     8521 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     8581 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     8641 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     8701 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     8761 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     8821 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     8881 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     8941 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     9001 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     9061 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     9121 ccccgaaaag tgccacctga c
//
