LOCUS       sequence-418250-        8426 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression in insect cells of SINTBAD-mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8426)
  AUTHORS   Adriaenssens E, Nguyen TN, Sawa-Makarska J, Khuu G, Schuschnig M, 
            Shoebridge S, Watts EM, Csalyi KD, Padman BS, Lazarou M, Martens S
  TITLE     Control of mitophagy initiation and progression by the TBK1 adaptors
            NAP1 and SINTBAD
  JOURNAL   bioRxiv September 25, 202
REFERENCE   2  (bases 1 to 8426)
  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: "bioRxiv 
            September 25, 202"
COMMENT     Sequence Label: pFastBac-Dual-GST-TEV-SINTBAD-mCherry
FEATURES             Location/Qualifiers
     source          1..8426
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..556
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(189..208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     398..419
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        582..686
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(687..755,756..1547)
                     /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(905..924)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1718..2306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2207..2226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2460..2477
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2611..2835
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2902..3435)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3624..3629,3630..3646,3647..3652))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    4160..4208
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4233..4252
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(4344..4453)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        4477..4568
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4493..4510
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4611..5264
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             5271..5288
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5298..5318
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             7176..7886
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(7321..7339)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7589..7608
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    8016..8150
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8053..8072)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8107..8126
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  8179..8344
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
       61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
      121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      361 ttttcgccct ttgacgttgg agtccacgtt cttaatagtg gactcttgtt ccaaactgga
      421 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
      481 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      541 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      601 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      661 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      721 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      781 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      841 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
      901 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
      961 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1021 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1081 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1141 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1201 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1261 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1321 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1381 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1441 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1501 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1561 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1621 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1861 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2341 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2401 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2461 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     2521 tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct ggcaaaatcg gttacggttg
     2581 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa
     2641 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga
     2701 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact
     2761 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg
     2821 gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc
     2881 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
     2941 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
     3001 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag
     3061 cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat
     3121 ctcactacgc ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc caacaaccgc
     3181 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta
     3241 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag
     3301 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat
     3361 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt
     3421 gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
     3481 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga
     3541 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     3601 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc
     3661 gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg
     3721 aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg
     3781 cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg cccttgcttc
     3841 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag
     3901 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct
     3961 atagttctag tggttggcta cgtacccgta gtggctatgg cagggcttgc cgccccgacg
     4021 ttggctgcga gccctgggcc ttcacccgaa cttgggggtt ggggtgggga aaaggaagaa
     4081 acgcgggcgt attggtccca atggggtctc ggtggggtat cgacagagtg ccagccctgg
     4141 gaccgaaccc cgcgtttatg aacaaacgac ccaacacccg tgcgttttat tctgtctttt
     4201 tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct
     4261 cccccatctc ccggtaccgc atgctatgca tcagctgcta gcaccatggc tcgagatccc
     4321 gggtgatcaa gtcttcgtcg agtgattgta aataaaatgt aatttacagt atagtatttt
     4381 aattaatata caaatgattt gataataatt cttatttaac tataatatat tgtgttgggt
     4441 tgaattaaag gtccgtatac tccggaatat taatagatca tggagataat taaaatgata
     4501 accatctcgc aaataaataa gtattttact gttttcgtaa cagttttgta ataaaaaaac
     4561 ctataaatat tccggattat tcataccgtc ccaccatcgg gcgcggatcc atgagtccta
     4621 tcctcggtta ctggaaaatc aaaggtctgg ttcagcccac ccgtctcctc ctggaatact
     4681 tggaagaaaa atacgaagag cacctgtacg agcgcgacga aggcgacaag tggcgtaaca
     4741 agaagttcga gctgggactg gaattcccca acctgccata ctacatcgac ggcgacgtca
     4801 agctgaccca gtccatggct atcatcagat acatcgccga caagcacaac atgctgggtg
     4861 gctgccctaa ggagcgcgct gaaatcagca tgctggaggg tgccgtgctg gacatccgtt
     4921 acggcgtctc aagaatcgct tactccaagg acttcgaaac tctgaaggtc gacttcctga
     4981 gcaagctgcc tgagatgctg aagatgttcg aagaccgtct gtgccacaag acctacctga
     5041 acggagacca cgtgactcac cccgacttca tgctgtacga cgctctggac gtggtcctgt
     5101 acatggaccc aatgtgcctg gacgctttcc ctaagctggt gtgcttcaag aagcgtatcg
     5161 aggccatccc tcagatcgac aagtacctga agtccagcaa gtacatcgct tggcctctgc
     5221 aaggatggca agctaccttc ggaggtggcg accaccctcc caagtcagac ctggtcccca
     5281 ggggttcccc agagttcgaa aacctgtact tccagggagg tggcatggag agcatgttcg
     5341 aagacgacat ctctatcctg actcaggagg ctctgggacc atctgaagtg tggctggaca
     5401 gccctggtga cccctctctg ggaggtgaca tgtgctccgc cagccacttc gctctgatca
     5461 ccgcctacgg cgacatcaag gagaggctgg gcggactgga gagagaaaac gccactctgc
     5521 gccgtaggct gaaggtgtac gagatcaagt accctctgat ctcagacttc ggcgaggaac
     5581 acggattcag cctgtacgaa atcaaggacg gatctctgct ggaggtggaa aaggtctcac
     5641 tgcagcagcg tctgaaccag ttccagcacg agctgcagaa gaacaaggaa caggaggaac
     5701 agctgggaga gatgatccag gcttacgaga agctgtgcgt cgaaaagtcc gacctggaga
     5761 ccgaactgag ggagatgaga gccctggtgg aaactcacct gcgccagatc tgcggcctgg
     5821 aacagcagct gcgtcagcag cagggactgc aggacgctgc cttctctaac ctctcaccac
     5881 ctcccgctcc tgccccacct tgcactgacc tggacctgca ctacctggct ctgcgcggtg
     5941 gctccggtct gtctcacgct ggttggcctg gcagcactcc ctctgtctca gacctggaga
     6001 gacgccgtct ggaggaagct ctggaggctg ctcagggtga agctcgcggc gctcagctga
     6061 gggaggaaca actccaggct gagtgcgaaa ggctgcaggg agagctgaag cagctgcagg
     6121 aaacccgtgc tcaggacctg gcctctaacc agtcagagag ggacatggct tgggtgaaga
     6181 gagtcggcga cgaccaggtg aacctggctc tggcctacac cgagctgact gaggaactgg
     6241 gtaggctgag agaactgtct tcactgcagg gccgcatcct gcgtactctg ctgcaggaac
     6301 aggccaggtc cggaggtcaa agacactccc ctctgagcca gcgtcactct cctgctcctc
     6361 agtgcccatc cccatcccct cctgctaggg ctgcccctcc ctgcccacct tgccagtcac
     6421 ctgtgcccca gaggagatcc ccagtgcctc cttgcccaag ccctcagcag aggcgttctc
     6481 cagcttctcc ttcatgcccc tccccagtgc ctcagaggag atcaccagtc cctccctctt
     6541 gccagtcccc aagcccacag aggcgtagcc ctgtcccacc ttcttgccct gctccccagc
     6601 ctcgtccccc acctccccca cctcccggag agcgcaccct ggccgaacgt gcttacgcca
     6661 agccaccttc tcaccacgtg aaggctggat tccagggtag gagatcttac tcagagctgg
     6721 ccgaaggagc tgcctacgct ggtgcttcac ccccatggct gcaggctgag gctgccactc
     6781 tgcctaagcc cagggcctac ggttccgaac tgtacggacc tggtagacca ctgagccctc
     6841 gccgtgcttt cgagggtatc cgcctgcgtt tcgaaaagca gccatctgag gaagacgagt
     6901 gggctgtgcc tacctctcct ccctcaccag aagtcggaac tatcagatgc gcttctttct
     6961 gcgccggttt cccaatccct gaatctcctg ctgctactgc ttacgctcac gctgaacacg
     7021 ctcagtcctg gccaagcatc aacctgctga tggagactgt cggcagcgac atcaggtctt
     7081 gccctctgtg ccagctggga ttcccagtgg gttaccctga cgacgccctg atcaagcaca
     7141 tcgactcaca cctggagaac tccaagatca agcttatggt gagcaagggc gaggaggata
     7201 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     7261 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     7321 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     7381 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     7441 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     7501 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     7561 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     7621 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     7681 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     7741 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     7801 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     7861 gcggcatgga cgagctgtac aagtagaagc ttgtcgagaa gtactagagg atcataatca
     7921 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     7981 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     8041 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     8101 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc tgatcactgc
     8161 ttgagcctag gagatccgaa ccagataagt gaaatctagt tccaaactat tttgtcattt
     8221 ttaattttcg tattagctta cgacgctaca cccagttccc atctattttg tcactcttcc
     8281 ctaaataatc cttaaaaact ccatttccac ccctcccagt tcccaactat tttgtccgcc
     8341 cacagcgggg catttttctt cctgttatgt ttttaatcaa acatcctgcc aactccatgt
     8401 gacaaaccgt catcttcggc tacttt
//
