LOCUS       sequence-448397-        9111 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 9111)
  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 9111)
  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-delta-206-230
FEATURES             Location/Qualifiers
     source          1..9111
                     /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             4807..4830
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    4864..5437
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4887..5439
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5031..5048)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5258..5277
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5439..6038
                     /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(5439..5458)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5935..5955
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6094..6682
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6147..6167)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6565..6576)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6685..6701)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6686..6702)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7211..7391
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7453..8041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7533..7552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8118..8909,8910..8978))
                     /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     8741..8760
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8979..9083)
                     /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 ttacagaatt aacttggcat tcatgtctac caggcaattt
     3901 gagaggaccc catacagaac tgctggtgag gacactggag taatataccc gtctgtagaa
     3961 gactctcaag aagtgtgtac cacctctttt tccacctccc caccatccca gctctcaagc
     4021 tctcgccttt cctatcagcc tgctgccctg ggcgttggat cagctgacct ggcttatcca
     4081 gtagtgtttg ctgctggctt aaatgctaca caccctcacc agctgatggt tcctgggaag
     4141 gaaggtgggg taccccttgc tcccaaccag cctgtccatg gtacccaggc tgaccaggag
     4201 agactggcaa cctgcacccc ttctgacaga acccactgtg ctgccacacc ctccagtagt
     4261 gaggatactg aaaccgtatc aaacagcagt gagggacggg cctcccctca cgatgtcttg
     4321 gagaccatct ttgtccgaaa agtgggggct tttgtcaaca aacccattaa ccaggtgacc
     4381 ctgacgagtt tggatatacc ctttgccatg tttgctccca agaatttgga gctggaggat
     4441 accgatccaa tggtgaatcc tccagattcc ccagagactg aatctcctct ccagggcagc
     4501 ctgcactcag atggctccag cgggggcagc agtggcaata cccatgatga ctttgttatg
     4561 atagacttta aaccagcttt ttctaaagat gacattcttc cgatggacct ggggaccttc
     4621 tatcgggagt ttcagaaccc acctcagctg agcagcctct ccatagatat tggagcacag
     4681 tccatggctg aagacttgga ctcattacca gagaagctgg ctgtgcatga gaagaatgtc
     4741 cgcgagtttg atgcctttgt ggaaaccctg cagtaagcta gcggaccgac gcgtacgcgg
     4801 ccgctcgatt acaaggatga cgacgataag gtttaaacct aggcgtagcg gccgcaaatt
     4861 ccgcccctct ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg
     4921 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
     4981 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     5041 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
     5101 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     5161 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     5221 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     5281 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     5341 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca
     5401 cggggacgtg gttttccttt gaaaaacacg atgataatat gaccgagtac aagcccacgg
     5461 tgcgcctcgc cacccgcgac gacgtccccc gggcagtacg caccctcgcc gccgcgttcg
     5521 ccgactaccc cgccacgcgc cacaccgtcg atccagaccg ccacatcgag cgggtcaccg
     5581 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     5641 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     5701 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     5761 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     5821 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     5881 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     5941 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     6001 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaac atgtttaagg gttccggttc
     6061 cactaggtac aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt
     6121 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     6181 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     6241 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     6301 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     6361 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     6421 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     6481 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     6541 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     6601 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     6661 tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gatcgagacc tagaaaaaca
     6721 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc
     6781 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac
     6841 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     6901 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta
     6961 cttccctgat tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg
     7021 atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga
     7081 gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt
     7141 attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc
     7201 ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     7261 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     7321 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     7381 aatctctagc agcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7441 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7501 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7561 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7621 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7681 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7741 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7801 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7861 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     7921 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7981 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     8041 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     8101 agggattttg gtcatgatta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8161 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8221 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8281 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8341 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8401 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8461 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8521 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8581 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8641 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8701 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8761 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8821 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8881 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8941 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9001 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9061 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga c
//
