LOCUS       sequence-448032-        6566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of recombinant protein for purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6566)
  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 6566)
  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: pCAG_GST-TEV-WIPI1
FEATURES             Location/Qualifiers
     source          1..6566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(163..179)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(163..179)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(176..198)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    187..203
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(211..217,218..235,236..241))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    256..277
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(406..423)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(577..1165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(657..676)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1336..2127,2128..2196))
                     /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     1959..1978
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2197..2301)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        2332..2711
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2713..2989
                     /label=chicken beta-actin promoter
     intron          2990..4006
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3930..3952
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4014..4033
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      4082..4091
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4088..4744
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MASPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
                     KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIR
                     YGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVV
                     LYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     4702..4724
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             4772..4792
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(6248..6267)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6313..6368
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6367..6386)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
ORIGIN
        1 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
       61 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta
      121 tggagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt
      181 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
      241 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
      301 tttccagtcg ggaaacctgt cgtgccagcg gatcgatccg ctgcattaat gaatcggcca
      361 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
      421 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      481 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      541 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      601 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      661 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      721 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      781 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      841 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      901 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
      961 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1021 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1081 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1141 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1201 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1261 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1321 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1381 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1441 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1501 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1561 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1621 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     1681 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1741 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1801 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1861 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1921 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     1981 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2041 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2101 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2161 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2221 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2281 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt cgacattgat
     2341 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     2401 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     2461 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     2521 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     2581 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     2641 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     2701 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct
     2761 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg
     2821 gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag
     2881 gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc
     2941 gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc
     3001 gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg
     3061 actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa
     3121 ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg
     3181 gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg
     3241 cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg
     3301 cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc
     3361 gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg
     3421 gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg
     3481 agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg gcgcggggct
     3541 cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg
     3601 ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg
     3661 cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc
     3721 tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg
     3781 gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc
     3841 gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg
     3901 ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc
     3961 tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt
     4021 gctggttatt gtgctgtctc atcattttgg caaagaattg cggccgtctc aggccgaatt
     4081 cgccaccatg gcatccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac
     4141 tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga
     4201 aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta
     4261 ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga
     4321 caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg
     4381 agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac
     4441 tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt
     4501 atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga
     4561 cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt
     4621 ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa
     4681 gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc
     4741 aaaatcggat gattacgata tcccaacgac cgaaaacctg tattttcagg gtgtcgacgg
     4801 taccatggaa gccgaagccg ctgatgctcc tcctggcgga gttgaatctg ccctgagctg
     4861 cttcagcttc aaccaggact gtaccagcct ggccacaggc acaaaggccg gctacaagct
     4921 gtttagcctg agcagcgtgg aacagctgga tcaggtgcac ggcagcaacg agatccccga
     4981 cgtgtacatc gtggaacggc tgttcagcag cagcctggtg gtggtcgtgt ctcacaccaa
     5041 gcctcggcag atgaacgtgt accacttcaa gaagggcacc gaaatctgca actacagcta
     5101 cagcagcaac atcctgagca tccggctgaa ccggcagaga ctgctcgtgt gtctggaaga
     5161 gagcatctac atccacaaca tcaaggacat gaagctgctc aagaccctgc tggacatccc
     5221 cgccaatcct acaggactgt gtgccctgtc catcaaccac agcaacagct acctggctta
     5281 ccccggctct ctgacatctg gcgagatcgt gctgtacgac ggcaacagcc tgaaaaccgt
     5341 gtgcacaatc gccgctcacg agggaacact ggccgccatc acctttaatg ccagcggctc
     5401 taaactggcc agcgccagcg aaaagggcac agtgatcaga gtgttcagcg tgcccgatgg
     5461 ccagaagctg tacgagtttc ggagaggcat gaagcgctac gtgaccatca gctccctggt
     5521 gttctctatg gacagccagt tcctgtgcgc cagctccaac acagagacag tgcacatctt
     5581 caagctggaa caagtgacca acagcagacc cgaggaaccc tctacttgga gcggctacat
     5641 gggcaagatg ttcatggccg ccaccaacta cctgcctaca caggtgtccg acatgatgca
     5701 ccaggaccgg gcctttgcca ccgccagact gaatttttcc ggccagcgga acatctgcac
     5761 cctgagcacc attcagaagc tgcccagact gctggtggcc tcttctagcg gccacctgta
     5821 catgtataat ctggaccctc aggacggcgg cgagtgcgtg ctgattaaga cacactctct
     5881 gctcggcagc ggcaccaccg aggaaaacaa agagaacgac ctgcggccta gcctgcctca
     5941 gtcttatgct gccacagtgg ccagacctag cgcctcttct gcttctacag tgcccggcta
     6001 tagcgaggat ggcggagcac ttagaggcga agtgatcccc gagcacgagt tcgctacagg
     6061 ccctgtctgt ctggacgacg agaacgagtt ccctcctatc atcctgtgcc ggggcaacca
     6121 gaagggcaag accaagcagt cttgactcga gactagtatc gcgataattc actcctcagg
     6181 tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc
     6241 actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
     6301 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     6361 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt
     6421 tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat
     6481 aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag
     6541 ccttgacttg aggttagatt tttttt
//
