LOCUS       sequence-437252-        6959 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Z-tagged proteins using Baculoviruses.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6959)
  TITLE     Mechanism of human PINK1 activation at the TOM complex in a 
            reconstituted system
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6959)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pFB-6HZB
FEATURES             Location/Qualifiers
     source          1..6959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(810..1255)
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     protein_bind    1270..1303
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1333..1353)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(1360..1530)
                     /codon_start=1
                     /product="Z domain of staphylococcal protein A, engineered 
                     to contain multiple positive charges (Hedhammar and Hober, 
                     2007)"
                     /label=Z-basic tag
                     /note="useful for purifying tagged proteins by 
                     cation-exchange chromatography"
                     /translation="VDNKFNKERRRARREIRHLPNLNREQRRAFIRSLRDDPSQSANLL
                     AEAKKLNDAQPK"
     CDS             complement(1540..1557)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(1619..1710)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(1677..1694)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     promoter        join(2062..2067,2068..2084,2085..2090)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2260..2793
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  2860..3084
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(3218..3235)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3389..3977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3469..3488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4148..4939,4940..5008))
                     /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     4771..4790
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5009..5113)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5139..5593)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5276..5297)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5487..5506
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     mobile_element  5777..5942
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    5971..6105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5995..6014)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6049..6068
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6347..6959,1..722,723..809))
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYKVPEFDSSTIKNISSAKGLDVWDSWPLQNTDGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
ORIGIN
        1 ccatcttcag ttccagtgtt tgcttcaaat actaagtatt tgtggccttt atcttctacg
       61 tagtgaggat ctctcagcgt atggttgtcg cctgagctgt agttgccttc atcgatgaac
      121 tgctgtacat tttgatacgt ttttccgtca ccgtcaaaga ttgatttata atcctctaca
      181 ccgttgatgt tcaaagagct gtctgatgct gatacgttaa cttgtgcagt tgtcagtgtt
      241 tgtttgccgt aatgtttacc ggagaaatca gtgtagaata aacggatttt tccgtcagat
      301 gtaaatgtgg ctgaacctga ccattcttgt gtttggtctt ttaggataga atcatttgca
      361 tcgaatttgt cgctgtcttt aaagacgcgg ccagcgtttt tccagctgtc aatagaagtt
      421 tcgccgactt tttgatagaa catgtaaatc gatgtgtcat ccgcattttt aggatctccg
      481 gctaatgcaa agacgatgtg gtagccgtga tagtttgcga cagtgccgtc agtgttttgt
      541 aatggccagc tgtcccaaac gtccaggcct tttgcagaag agatattttt aattgtggac
      601 gaatcgaact caggaacttt atatttttca tttttttgct gttcagggat ttgcagcata
      661 tcatggcgtg taatatggga aatgccgtat gtttccttat atggcttttg gttcgtttct
      721 ttcgcaaacg cttgagttgc gcctcctgcc agcagtgcgg tagtaaaggt taatactgtt
      781 gcttgttttg caaacttttt gatgttcatc gttcatgtct ccttttttat gtactgtgtt
      841 agcggtctgc ttcttccagc cctcctgttt gaagatggca agttagttac gcacaataaa
      901 aaaagaccta aaatatgtaa ggggtgacgc caaagtatac actttgccct ttacacattt
      961 taggtcttgc ctgctttatc agtaacaaac ccgcgcgatt tacttttcga cctcattcta
     1021 ttagactctc gtttggattg caactggtct attttcctct tttgtttgat agaaaatcat
     1081 aaaaggattt gcagactacg ggcctaaaga actaaaaaat ctatctgttt cttttcattc
     1141 tctgtatttt ttatagtttc tgttgcatgg gcataaagtt gcctttttaa tcacaattca
     1201 gaaaatatca taatatctca tttcactaaa taatagtgaa cggcaggtat atgtggacgt
     1261 cgcggccgca taacttcgta taatgtatgc tatacgaagt tatggatctt tcaggaggag
     1321 aagctagctt atggattgga agtacaggtt ctctgtaccc ttcggctgcg catcgttcag
     1381 cttcttcgct tccgccagca ggttcgcgct ctggctcgga tcatcgcgca ggctgcgaat
     1441 aaatgcgcga cgctgttcgc ggttcaggtt cggcagatgg cgaatttcac ggcgagctct
     1501 tcgacgctcc ttgttgaact tgttatccac accagaagaa tgatgatgat gatggtggcc
     1561 catggttttc ggaccgagat ccgcgcccga tggtgggacg gtatgaataa tccggaatat
     1621 ttataggttt ttttattaca aaactgttac gaaaacagta aaatacttat ttatttgcga
     1681 gatggttatc attttaatta tctccatgat ctattaatat tccggagtat acgtagccaa
     1741 ccactagaac tatagctaga gtcctgggcg aacaaacgat gctcgccttc cagaaaaccg
     1801 aggatgcgaa ccacttcatc cggggtcagc accaccggca agcgccgcga cggccgaggt
     1861 cttccgatct cctgaagcca gggcagatcc gtgcacagca ccttgccgta gaagaacagc
     1921 aaggccgcca atgcctgacg atgcgtggag accgaaacct tgcgctcgtt cgccagccag
     1981 gacagaaatg cctcgacttc gctgctgccc aaggttgccg ggtgacgcac accgtggaaa
     2041 cggatgaagg cacgaaccca gttgacataa gcctgttcgg ttcgtaaact gtaatgcaag
     2101 tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc
     2161 gcagtggcgg ttttcatggc ttgttatgac tgtttttttg tacagtctat gcctcgggca
     2221 tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat
     2281 gttacgcagc agggcagtcg ccctaaaaca aagttaggtg gctcaagtat gggcatcatt
     2341 cgcacatgta ggctcggccc tgaccaagtc aaatccatgc gggctgctct tgatcttttc
     2401 ggtcgtgagt tcggagacgt agccacctac tcccaacatc agccggactc cgattacctc
     2461 gggaacttgc tccgtagtaa gacattcatc gcgcttgctg ccttcgacca agaagcggtt
     2521 gttggcgctc tcgcggctta cgttctgccc aagtttgagc agccgcgtag tgagatctat
     2581 atctatgatc tcgcagtctc cggcgagcac cggaggcagg gcattgccac cgcgctcatc
     2641 aatctgctca agcatgaggc caacgcgctt ggtgcttatg tgatctacgt gcaagcagat
     2701 tacggtgacg atcccgcagt ggctctctat acaaagttgg gcatacggga agaagtgatg
     2761 cactttgata tcgacccaag taccgccacc taacaattcg ttcaagccga gatcggcttc
     2821 ccggccgcgg agttgttcgg taaattgtca caacgccgcg aatatagtct ttaccatgcc
     2881 cttggccacg cccctcttta atacgacggg caatttgcac ttcagaaaat gaagagtttg
     2941 ctttagccat aacaaaagtc cagtatgctt tttcacagca taactggact gatttcagtt
     3001 tacaactatt ctgtctagtt taagacttta ttgtcatagt ttagatctat tttgttcagt
     3061 ttaagacttt attgtccgcc cacacccgct tacgcagggc atccatttat tactcaaccg
     3121 taaccgattt tgccaggtta cgcggctggt ctatgcggtg tgaaataccg cacagatgcg
     3181 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3241 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3301 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3361 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3421 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3481 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3541 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3601 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3661 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3721 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3781 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3841 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3901 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3961 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4021 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4081 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4141 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4201 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4261 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4321 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4381 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4441 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4501 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4561 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4621 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4681 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4741 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4801 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4861 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4921 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4981 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5041 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5101 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     5161 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     5221 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     5281 ttggaacaag agtccactat taagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     5341 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     5401 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     5461 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     5521 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     5581 ctacagggcg cgtcccattc gccattcagg ctgcaaataa gcgttgatat tcagtcaatt
     5641 acaaacatta ataacgaaga gatgacagaa aaattttcat tctgtgacag agaaaaagta
     5701 gccgaagatg acggtttgtc acatggagtt ggcaggatgt ttgattaaaa acataacagg
     5761 aagaaaaatg ccccgctgtg ggcggacaaa atagttggga actgggaggg gtggaaatgg
     5821 agtttttaag gattatttag ggaagagtga caaaatagat gggaactggg tgtagcgtcg
     5881 taagctaata cgaaaattaa aaatgacaaa atagtttgga actagatttc acttatctgg
     5941 ttcggatctc ctaggcgata tcagtgatca gatccagaca tgataagata cattgatgag
     6001 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     6061 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     6121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     6181 ctctacaaat gtggtatggc tgattatgat cctctagtac ttctcgacaa gcttgtcgac
     6241 ggagctcgaa ttcggatccg tatccacctt tacttactag taaactcctc gaaaagtgcc
     6301 acctgacgtc aatgccaata ggatatcggc attttctttt gcgtttttat ttgttaactg
     6361 ttaattgtcc ttgttcaagg atgctgtctt tgacaacaga tgttttcttg cctttgatgt
     6421 tcagcaggaa gctaggcgca aacgttgatt gtttgtctgc gtagaatcct ctgtttgtca
     6481 tatagcttgt aatcacgaca ttgtttcctt tcgcttgagg tacagcgaag tgtgagtaag
     6541 taaaggttac atcgttagga tcaagatcca tttttaacac aaggccagtt ttgttcagcg
     6601 gcttgtatgg gccagttaaa gaattagaaa cataaccaag catgtaaata tcgttagacg
     6661 taatgccgtc aatcgtcatt tttgatccgc gggagtcagt gaacaggtac catttgccgt
     6721 tcattttaaa gacgttcgcg cgttcaattt catctgttac tgtgttagat gcaatcagcg
     6781 gtttcatcac ttttttcagt gtgtaatcat cgtttagctc aatcataccg agagcgccgt
     6841 ttgctaactc agccgtgcgt tttttatcgc tttgcagaag tttttgactt tcttgacgga
     6901 agaatgatgt gcttttgcca tagtatgctt tgttaaataa agattcttcg ccttggtag
//
