LOCUS       sequence-391287-        8438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for the expression and purification of EGFP labelled SINTBAD 
            (SMC Internal No. 1613).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8438)
  AUTHORS   Nguyen TN, Sawa-Makarska J, Khuu G, Lam WK, Adriaenssens E, 
            Fracchiolla D, Shoebridge S, Bernklau D, Padman BS, Skulsuppaisarn 
            M, Lindblom RSJ, Martens S, Lazarou M
  TITLE     Unconventional initiation of PINK1/Parkin mitophagy by Optineurin.
  JOURNAL   Mol Cell. 2023 May 18;83(10):1693-1709.e9. doi: 
            10.1016/j.molcel.2023.04.021.
  PUBMED    37207627
REFERENCE   2  (bases 1 to 8438)
  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.2023.04"; journalName: "Mol Cell"; date: 
            "2023-05-18- 18"; volume: "83"; issue: "10"; pages: "1693-1709.e9"
COMMENT     Sequence Label: pFastBac_Dual-GST-TEV-SINTBAD-EGFP (codon opti...
FEATURES             Location/Qualifiers
     source          1..8438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(102..107,108..124,125..130))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    638..686
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     711..730
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(822..931)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        955..1046
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     971..988
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             1089..1742
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             1749..1766
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1776..1796
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3660..4373
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     polyA_signal    4506..4640
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4543..4562)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4597..4616
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4669..4834
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      5018..5472
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5105..5124)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5314..5335
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5498..5602
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5603..5671,5672..6463)
                     /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     complement(5821..5840)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6634..7222
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7123..7142
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7376..7393
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  7527..7751
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(7818..8351)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 agtcataaca agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg
       61 ttctggacca gttgcgtgag cgcatacgct acttgcatta cagtttacga accgaacagg
      121 cttatgtcaa ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct
      181 tgggcagcag cgaagtcgag gcatttctgt cctggctggc gaacgagcgc aaggtttcgg
      241 tctccacgca tcgtcaggca ttggcggcct tgctgttctt ctacggcaag gtgctgtgca
      301 cggatctgcc cttgcttcag gagatcggta gacctcggcc gtcgcggcgc ttgccggtgg
      361 tgctgacccc ggatgaagtg gttcgcatcc tcggttttct ggaaggcgag catcgtttgt
      421 tcgcccagga ctctagctat agttctagtg gttggctacg tacccgtagt ggctatggca
      481 gggcttgccg ccccgacgtt ggctgcgagc cctgggcctt cacccgaact tgggggttgg
      541 ggtggggaaa aggaagaaac gcgggcgtat tggtcccaat ggggtctcgg tggggtatcg
      601 acagagtgcc agccctggga ccgaaccccg cgtttatgaa caaacgaccc aacacccgtg
      661 cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt
      721 ccgtgtttca gttagcctcc cccatctccc ggtaccgcat gctatgcatc agctgctagc
      781 accatggctc gagatcccgg gtgatcaagt cttcgtcgag tgattgtaaa taaaatgtaa
      841 tttacagtat agtattttaa ttaatataca aatgatttga taataattct tatttaacta
      901 taatatattg tgttgggttg aattaaaggt ccgtatactc cggaatatta atagatcatg
      961 gagataatta aaatgataac catctcgcaa ataaataagt attttactgt tttcgtaaca
     1021 gttttgtaat aaaaaaacct ataaatattc cggattattc ataccgtccc accatcgggc
     1081 gcggatccat gagtcctatc ctcggttact ggaaaatcaa aggtctggtt cagcccaccc
     1141 gtctcctcct ggaatacttg gaagaaaaat acgaagagca cctgtacgag cgcgacgaag
     1201 gcgacaagtg gcgtaacaag aagttcgagc tgggactgga attccccaac ctgccatact
     1261 acatcgacgg cgacgtcaag ctgacccagt ccatggctat catcagatac atcgccgaca
     1321 agcacaacat gctgggtggc tgccctaagg agcgcgctga aatcagcatg ctggagggtg
     1381 ccgtgctgga catccgttac ggcgtctcaa gaatcgctta ctccaaggac ttcgaaactc
     1441 tgaaggtcga cttcctgagc aagctgcctg agatgctgaa gatgttcgaa gaccgtctgt
     1501 gccacaagac ctacctgaac ggagaccacg tgactcaccc cgacttcatg ctgtacgacg
     1561 ctctggacgt ggtcctgtac atggacccaa tgtgcctgga cgctttccct aagctggtgt
     1621 gcttcaagaa gcgtatcgag gccatccctc agatcgacaa gtacctgaag tccagcaagt
     1681 acatcgcttg gcctctgcaa ggatggcaag ctaccttcgg aggtggcgac caccctccca
     1741 agtcagacct ggtccccagg ggttccccag agttcgaaaa cctgtacttc cagggaggtg
     1801 gcatggagag catgttcgaa gacgacatct ctatcctgac tcaggaggct ctgggaccat
     1861 ctgaagtgtg gctggacagc cctggtgacc cctctctggg aggtgacatg tgctccgcca
     1921 gccacttcgc tctgatcacc gcctacggcg acatcaagga gaggctgggc ggactggaga
     1981 gagaaaacgc cactctgcgc cgtaggctga aggtgtacga gatcaagtac cctctgatct
     2041 cagacttcgg cgaggaacac ggattcagcc tgtacgaaat caaggacgga tctctgctgg
     2101 aggtggaaaa ggtctcactg cagcagcgtc tgaaccagtt ccagcacgag ctgcagaaga
     2161 acaaggaaca ggaggaacag ctgggagaga tgatccaggc ttacgagaag ctgtgcgtcg
     2221 aaaagtccga cctggagacc gaactgaggg agatgagagc cctggtggaa actcacctgc
     2281 gccagatctg cggcctggaa cagcagctgc gtcagcagca gggactgcag gacgctgcct
     2341 tctctaacct ctcaccacct cccgctcctg ccccaccttg cactgacctg gacctgcact
     2401 acctggctct gcgcggtggc tccggtctgt ctcacgctgg ttggcctggc agcactccct
     2461 ctgtctcaga cctggagaga cgccgtctgg aggaagctct ggaggctgct cagggtgaag
     2521 ctcgcggcgc tcagctgagg gaggaacaac tccaggctga gtgcgaaagg ctgcagggag
     2581 agctgaagca gctgcaggaa acccgtgctc aggacctggc ctctaaccag tcagagaggg
     2641 acatggcttg ggtgaagaga gtcggcgacg accaggtgaa cctggctctg gcctacaccg
     2701 agctgactga ggaactgggt aggctgagag aactgtcttc actgcagggc cgcatcctgc
     2761 gtactctgct gcaggaacag gccaggtccg gaggtcaaag acactcccct ctgagccagc
     2821 gtcactctcc tgctcctcag tgcccatccc catcccctcc tgctagggct gcccctccct
     2881 gcccaccttg ccagtcacct gtgccccaga ggagatcccc agtgcctcct tgcccaagcc
     2941 ctcagcagag gcgttctcca gcttctcctt catgcccctc cccagtgcct cagaggagat
     3001 caccagtccc tccctcttgc cagtccccaa gcccacagag gcgtagccct gtcccacctt
     3061 cttgccctgc tccccagcct cgtcccccac ctcccccacc tcccggagag cgcaccctgg
     3121 ccgaacgtgc ttacgccaag ccaccttctc accacgtgaa ggctggattc cagggtagga
     3181 gatcttactc agagctggcc gaaggagctg cctacgctgg tgcttcaccc ccatggctgc
     3241 aggctgaggc tgccactctg cctaagccca gggcctacgg ttccgaactg tacggacctg
     3301 gtagaccact gagccctcgc cgtgctttcg agggtatccg cctgcgtttc gaaaagcagc
     3361 catctgagga agacgagtgg gctgtgccta cctctcctcc ctcaccagaa gtcggaacta
     3421 tcagatgcgc ttctttctgc gccggtttcc caatccctga atctcctgct gctactgctt
     3481 acgctcacgc tgaacacgct cagtcctggc caagcatcaa cctgctgatg gagactgtcg
     3541 gcagcgacat caggtcttgc cctctgtgcc agctgggatt cccagtgggt taccctgacg
     3601 acgccctgat caagcacatc gactcacacc tggagaactc caagatcggc ggaggtggca
     3661 tgtccaaggg cgaggaactg ttcaccggag tggtccccat cctggtggaa ctggacggcg
     3721 acgtcaacgg acacaagttc tccgtgagcg gcgagggaga aggtgacgct acctacggca
     3781 agctgactct gaagttcatc tgcaccactg gcaagctgcc cgtgccatgg cctaccctgg
     3841 tcaccactct gacttacggt gtccagtgct tctctcgcta ccccgaccac atgaagcagc
     3901 acgacttctt caagtcagct atgccagagg gctacgtgca ggaaaggacc atcttcttca
     3961 aggacgacgg aaactacaag accagagccg aggtgaagtt cgaaggagac actctggtca
     4021 accgcatcga gctgaagggt atcgacttca aggaagacgg taacatcctg ggccacaagc
     4081 tggagtacaa ctacaactcc cacaacgtgt acatcatggc tgacaagcag aagaacggta
     4141 tcaaggtcaa cttcaagatc cgtcacaaca tcgaagacgg cagcgtgcag ctggccgacc
     4201 actaccagca gaacactcct atcggcgacg gacctgtcct gctgccagac aaccactacc
     4261 tgtctactca gtcagctctg tccaaggacc ctaacgagaa gcgcgaccac atggtgctgt
     4321 tggagttcgt tacggctgct ggaatcaccc acggtatgga tgagctctac aaataaaagc
     4381 ttgtcgagaa gtactagagg atcataatca gccataccac atttgtagag gttttacttg
     4441 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     4501 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4561 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4621 tatcttatca tgtctggatc tgatcactgc ttgagcctag gagatccgaa ccagataagt
     4681 gaaatctagt tccaaactat tttgtcattt ttaattttcg tattagctta cgacgctaca
     4741 cccagttccc atctattttg tcactcttcc ctaaataatc cttaaaaact ccatttccac
     4801 ccctcccagt tcccaactat tttgtccgcc cacagcgggg catttttctt cctgttatgt
     4861 ttttaatcaa acatcctgcc aactccatgt gacaaaccgt catcttcggc tactttttct
     4921 ctgtcacaga atgaaaattt ttctgtcatc tcttcgttat taatgtttgt aattgactga
     4981 atatcaacgc ttatttgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     5041 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     5101 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     5161 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5221 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5281 cgccctttga cgttggagtc cacgttctta atagtggact cttgttccaa actggaacaa
     5341 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     5401 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     5461 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5521 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5581 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5641 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5701 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5761 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5821 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5881 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5941 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6001 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6061 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6121 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     6181 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6241 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6301 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6361 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6421 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6481 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6541 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6601 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6661 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6721 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6781 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6841 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6901 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6961 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7021 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7081 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7141 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7201 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7261 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     7321 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     7381 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     7441 cggtatttca caccgcatag accagccgcg taacctggca aaatcggtta cggttgagta
     7501 ataaatggat gccctgcgta agcgggtgtg ggcggacaat aaagtcttaa actgaacaaa
     7561 atagatctaa actatgacaa taaagtctta aactagacag aatagttgta aactgaaatc
     7621 agtccagtta tgctgtgaaa aagcatactg gacttttgtt atggctaaag caaactcttc
     7681 attttctgaa gtgcaaattg cccgtcgtat taaagagggg cgtggccaag ggcatggtaa
     7741 agactatatt cgcggcgttg tgacaattta ccgaacaact ccgcggccgg gaagccgatc
     7801 tcggcttgaa cgaattgtta ggtggcggta cttgggtcga tatcaaagtg catcacttct
     7861 tcccgtatgc ccaactttgt atagagagcc actgcgggat cgtcaccgta atctgcttgc
     7921 acgtagatca cataagcacc aagcgcgttg gcctcatgct tgaggagatt gatgagcgcg
     7981 gtggcaatgc cctgcctccg gtgctcgccg gagactgcga gatcatagat atagatctca
     8041 ctacgcggct gctcaaactt gggcagaacg taagccgcga gagcgccaac aaccgcttct
     8101 tggtcgaagg cagcaagcgc gatgaatgtc ttactacgga gcaagttccc gaggtaatcg
     8161 gagtccggct gatgttggga gtaggtggct acgtctccga actcacgacc gaaaagatca
     8221 agagcagccc gcatggattt gacttggtca gggccgagcc tacatgtgcg aatgatgccc
     8281 atacttgagc cacctaactt tgttttaggg cgactgccct gctgcgtaac atcgttgctg
     8341 ctgcgtaaca tcgttgctgc tccataacat caaacatcga cccacggcgt aacgcgcttg
     8401 ctgcttggat gcccgaggca tagactgtac aaaaaaac
//
