LOCUS       sequence-324118-        6581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transfer vector for multigene expression to generate recombinant 
            baculoviruses by homologous recombination. Contains a p10/pH dual 
            expression cassette with a C-ter 10-His tag under the pH promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6581)
  AUTHORS   Kolesnikova O, Zachayus A, Pichard S, Osz J, Rochel N, Rossolillo P,
            Kolb-Cheynel I, Troffer-Charlier N, Compe E, Bensaude O, Berger I, 
            Poterszman A
  TITLE     HR-Bac, a toolbox based on homologous recombination for expression, 
            screening and production of multiprotein complexes using the 
            baculovirus expression system.
  JOURNAL   Sci Rep. 2022 Feb 7;12(1):2030. doi: 10.1038/s41598-021-04715-5.
  PUBMED    35132103
REFERENCE   2  (bases 1 to 6581)
  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: "Sci Rep."; 
            date: "2022-02-7"; volume: "12(1):2030. doi"; pages: " 
            10.1038/s41598-021-04715-5"
COMMENT     Sequence Label: pAC8_MF-PH-3C-10His-Cter (VE5622)
FEATURES             Location/Qualifiers
     source          1..6581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     330..1157
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             330..454
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(492..1097)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     protein_bind    complement(1164..1197)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1387..1435
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1460..1479
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(1571..1680)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        1725..1816
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1741..1758
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             1931..1954
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1955..1984
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     polyA_signal    2130..2264
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2167..2186)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2221..2240
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_recomb     2383..3740
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(2399..3740)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="PSLRQSLYNTIAYIERLLNIGTVNDSEITMLIADFYDLYSNYNIE
                     LPPPQALPRSRRPSVVQPAAPAPVPTIVREQTKPEQIIPAAPPPPPSPVPNIPAPPPPP
                     PPSMSELPPAPPMPTEPQPAAPLDDRQQLLEAIRNEKNRTRLRPVKPKTAPETSTIVEV
                     PTVLPKETFEPKPPSASPPPPPPPPPPPAPPAPPPMVDLSSAPPPPPLVDLPSEMLPPP
                     APSLSNVLSELKSGTVRLKPAQKRPQSEIIPKSSTTNLIADVLADTINRRRVAMAKSSS
                     EATSNDEGWDDDDNRPNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVE
                     PLLQNKTQTNIDKARLLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDL
                     IFKFRYKDAENHLIFALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     rep_origin      3887..4342
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3974..3993)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4184..4205
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4359..4378)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4478..4500
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4538..4556)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4624..4728
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4729..4797,4798..5589)
                     /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(4947..4966)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5760..6348
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6249..6268
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6502..6519
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cagaacggct ccgcccacta ttaatgaaat taaaaattcc aattttaaaa aacgcagcaa
       61 gagaaacatt tgtatgaaag aatgcgtaga aggaaagaaa aatgtcgtcg acatgctgaa
      121 caacaagatt aatatgcctc cgtgtataaa aaaaatattg aacgatttga aagaaaacaa
      181 tgtaccgcgc ggcggtatgt acaggaagag gtttatacta aactgttaca ttgcaaacgt
      241 ggtttcgtgt gccaagtgtg aaaaccgatg tttaatcaag gctctgacgc atttctacaa
      301 ccacgactcc aagtgtgtgg gtgaagtcat gcatctttta atcaaatccc aagatgtgta
      361 taaaccacca aactgccaaa aaatgaaaac tgtcgacaag ctctgtccgt ttgctggcaa
      421 ctgcaagggt ctcaatccta tttgtaatta ttgaataata aaacaattat aaatgtcaaa
      481 tttgtttttt attaacgata caaaccaaac gcaacaagaa catttgtagt attatctata
      541 attgaaaacg cgtagttata atcgctgagg taatatttaa aatcattttc aaatgattca
      601 cagttaattt gcgacaatat aattttattt tcacataaac tagacgcctt gtcgtcttct
      661 tcttcgtatt ccttctcttt ttcatttttc tcctcataaa aattaacata gttattatcg
      721 tatccatata tgtatctatc gtatagagta aattttttgt tgtcataaat atatatgtct
      781 tttttaatgg ggtgtatagt accgctgcgc atagtttttc tgtaatttac aacagtgcta
      841 ttttctggta gttcttcgga gtgtgttgct ttaattatta aatttatata atcaatgaat
      901 ttgggatcgt cggttttgta caatatgttg ccggcatagt acgcagcttc ttctagttca
      961 attacaccat tttttagcag caccggatta acataacttt ccaaaatgtt gtacgaaccg
     1021 ttaaacaaaa acagttcacc tcccttttct atactattgt ctgcgagcag ttgtttgttg
     1081 ttaaaaataa cagccattgt aatgagacgc acaaactaat atcacaaact ggaaatgtct
     1141 atcaatatat agttgctgat atcataactt cgtatagcat acattatacg aagttatctg
     1201 gtttaaacgt acccgtagtg gctatggcag ggcttgccgc cccgacgttg gctgcgagcc
     1261 ctgggccttc acccgaactt gggggttggg gtggggaaaa ggaagaaacg cgggcgtatt
     1321 ggtcccaatg gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc
     1381 gtttatgaac aaacgaccca acacccgtgc gttttattct gtctttttat tgccgtcata
     1441 gcgcgggttc cttccggtat tgtctccttc cgtgtttcag ttagcctccc ccatctcccg
     1501 gtaccgcatg ctatgcatca gctgctagca ccatggctcg agatcccggg tgatcaagtc
     1561 ttcgtcgagt gattgtaaat aaaatgtaat ttacagtata gtattttaat taatatacaa
     1621 atgatttgat aataattctt atttaactat aatatattgt gttgggttga attaaaggtc
     1681 cgtatactag tatcgattcg cgacctactc cggaatatta atagatcatg gagataatta
     1741 aaatgataac catctcgcaa ataaataagt attttactgt tttcgtaaca gttttgtaat
     1801 aaaaaaacct ataaatattc cggattattc ataccgtccc accatcgggc gcggatcccg
     1861 gtccgaagcg cgcggaattc aaaggcctac gtcgacgagc tcactagtcg cggccgcttt
     1921 cgaatctaga ctggaagttc tgtttcaggg tccgcaccat catcaccatc accatcacca
     1981 ccattaactg cagtctcgac aagcttgtcg agaagtacta gaggatcata atcagccata
     2041 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     2101 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     2161 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2221 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgatca ctgcttgagc
     2281 ctagaagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     2341 gagcaataac tatcataacc cctaggcgat taattgatcc gggttattag tacatttatt
     2401 aagcgctaga ttctgtgcgt tgttgattta cagacaattg ttgtacgtat tttaataatt
     2461 cattaaattt ataatcttta gggtggtatg ttagagcgaa aatcaaatga ttttcagcgt
     2521 ctttatatct gaatttaaat attaaatcct caatagattt gtaaaatagg tttcgattag
     2581 tttcaaacaa gggttgtttt tccgaaccga tggctggact atctaatgga ttttcgctca
     2641 acgccacaaa acttgccaaa tcttgtagca gcaatctagc tttgtcgata ttcgtttgtg
     2701 ttttgttttg taataaaggt tcgacgtcgt tcaaaatatt atgcgctttt gtatttcttt
     2761 catcactgtc gttagtgtac aattgactcg acgtaaacac gttaaataaa gcttggacat
     2821 atttaacatc gggcgtgtta gctttattag gccgattatc gtcgtcgtcc caaccctcgt
     2881 cgttagaagt tgcttccgaa gacgattttg ccatagccac acgacgccta ttaattgtgt
     2941 cggctaacac gtccgcgatc aaatttgtag ttgagctttt tggaattatt tctgattgcg
     3001 ggcgtttttg ggcgggtttc aatctaactg tgcccgattt taattcagac aacacgttag
     3061 aaagcgatgg tgcaggcggt ggtaacattt cagacggcaa atctactaat ggcggcggtg
     3121 gtggagctga tgataaatct accatcggtg gaggcgcagg cggggctggc ggcggaggcg
     3181 gaggcggagg tggtggcggt gatgcagacg gcggtttagg ctcaaatgtc tctttaggca
     3241 acacagtcgg cacctcaact attgtactgg tttcgggcgc cgtttttggt ttgaccggtc
     3301 tgagacgagt gcgatttttt tcgtttctaa tagcttccaa caattgttgt ctgtcgtcta
     3361 aaggtgcagc gggttgaggt tccgtcggca ttggtggagc gggcggcaat tcagacatcg
     3421 atggtggtgg tggtggtgga ggcgctggaa tgttaggcac gggagaaggt ggtggcggcg
     3481 gtgccgccgg tataatttgt tctggtttag tttgttcgcg cacgattgtg ggcaccggcg
     3541 caggcgccgc tggctgcaca acggaaggtc gtctgcttcg aggcagcgct tggggtggtg
     3601 gcaattcaat attataattg gaatacaaat cgtaaaaatc tgctataagc attgtaattt
     3661 cgctatcgtt taccgtgccg atatttaaca accgctcaat gtaagcaatt gtattgtaaa
     3721 gagattgtct caagctcgga tcgatcccgc acgccgataa caagcctttt catttttact
     3781 acagcattgt agtggcgaga cacttcgctg tcgtcgcctg atgcggtatt ttctccttac
     3841 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     3901 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3961 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4021 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4081 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     4141 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4201 actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg
     4261 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     4321 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     4381 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4441 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4501 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     4561 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     4621 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4681 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4741 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4801 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4861 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4921 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4981 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5041 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5101 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5161 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5221 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5281 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5341 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5401 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5461 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5521 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5581 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5641 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5701 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5761 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5821 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5881 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5941 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6001 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6061 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6121 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6181 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6241 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6301 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6361 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6421 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     6481 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     6541 cgcaaaccgc ctctccccgc gcgttggccg attcattaat g
//
