LOCUS       sequence-324157-        6541 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 an C-ter10His tag under the pH promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6541)
  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 6541)
  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-p10-10His-Cter (VE5742)
FEATURES             Location/Qualifiers
     source          1..6541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(275..366)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(333..350)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     promoter        411..520
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             568..597
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(627..646)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    671..719
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    909..942
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_recomb     949..1776
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             1009..1614
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     CDS             complement(1652..1776)
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     primer_bind     complement(2168..2185)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2339..2927)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2419..2438)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3098..3889,3890..3958))
                     /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     3721..3740
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3959..4063)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4131..4149
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4187..4209)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4309..4328
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4345..4800)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4482..4503)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4694..4713
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     misc_recomb     4947..6304
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             4947..6288
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="PSLRQSLYNTIAYIERLLNIGTVNDSEITMLIADFYDLYSNYNIE
                     LPPPQALPRSRRPSVVQPAAPAPVPTIVREQTKPEQIIPAAPPPPPSPVPNIPAPPPPP
                     PPSMSELPPAPPMPTEPQPAAPLDDRQQLLEAIRNEKNRTRLRPVKPKTAPETSTIVEV
                     PTVLPKETFEPKPPSASPPPPPPPPPPPAPPAPPPMVDLSSAPPPPPLVDLPSEMLPPP
                     APSLSNVLSELKSGTVRLKPAQKRPQSEIIPKSSTTNLIADVLADTINRRRVAMAKSSS
                     EATSNDEGWDDDDNRPNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVE
                     PLLQNKTQTNIDKARLLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDL
                     IFKFRYKDAENHLIFALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     polyA_signal    join(6423..6541,1..16)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6447..6466)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6501..6520
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
       61 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga
      121 tcctctagta cttctcgaca agcttgtcga gactgcaggc tctagattcg aaagcggccg
      181 cgactagtga gctcgtcgac gtaggccttt gaattccgcg cgcttcggac cgggatccgc
      241 gcccgatggt gggacggtat gaataatccg gaatatttat aggttttttt attacaaaac
      301 tgttacgaaa acagtaaaat acttatttat ttgcgagatg gttatcattt taattatctc
      361 catgatctat taatattccg gagtaggtcg cgaatcgata ctagtatacg gacctttaat
      421 tcaacccaac acaatatatt atagttaaat aagaattatt atcaaatcat ttgtatatta
      481 attaaaatac tatactgtaa attacatttt atttacaatc actcgacgaa gacttgatca
      541 cccgggatct cgagccatgg tgctagccac catcatcacc atcaccatca ccaccattaa
      601 ggtaccggga gatgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
      661 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca
      721 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
      781 ggaccaatac gcccgcgttt cttccttttc cccaccccaa cccccaagtt cgggtgaagg
      841 cccagggctc gcagccaacg tcggggcggc aagccctgcc atagccacta cgggtacgtt
      901 taaaccagat aacttcgtat aatgtatgct atacgaagtt atgatatcag caactatata
      961 ttgatagaca tttccagttt gtgatattag tttgtgcgtc tcattacaat ggctgttatt
     1021 tttaacaaca aacaactgct cgcagacaat agtatagaaa agggaggtga actgtttttg
     1081 tttaacggtt cgtacaacat tttggaaagt tatgttaatc cggtgctgct aaaaaatggt
     1141 gtaattgaac tagaagaagc tgcgtactat gccggcaaca tattgtacaa aaccgacgat
     1201 cccaaattca ttgattatat aaatttaata attaaagcaa cacactccga agaactacca
     1261 gaaaatagca ctgttgtaaa ttacagaaaa actatgcgca gcggtactat acaccccatt
     1321 aaaaaagaca tatatattta tgacaacaaa aaatttactc tatacgatag atacatatat
     1381 ggatacgata ataactatgt taatttttat gaggagaaaa atgaaaaaga gaaggaatac
     1441 gaagaagaag acgacaaggc gtctagttta tgtgaaaata aaattatatt gtcgcaaatt
     1501 aactgtgaat catttgaaaa tgattttaaa tattacctca gcgattataa ctacgcgttt
     1561 tcaattatag ataatactac aaatgttctt gttgcgtttg gtttgtatcg ttaataaaaa
     1621 acaaatttga catttataat tgttttatta ttcaataatt acaaatagga ttgagaccct
     1681 tgcagttgcc agcaaacgga cagagcttgt cgacagtttt cattttttgg cagtttggtg
     1741 gtttatacac atcttgggat ttgattaaaa gatgcatgac ttcacccaca cacttggagt
     1801 cgtggttgta gaaatgcgtc agagccttga ttaaacatcg gttttcacac ttggcacacg
     1861 aaaccacgtt tgcaatgtaa cagtttagta taaacctctt cctgtacata ccgccgcgcg
     1921 gtacattgtt ttctttcaaa tcgttcaata ttttttttat acacggaggc atattaatct
     1981 tgttgttcag catgtcgacg acatttttct ttccttctac gcattctttc atacaaatgt
     2041 ttctcttgct gcgtttttta aaattggaat ttttaatttc attaatagtg ggcggagccg
     2101 ttctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2161 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2221 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2281 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2341 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2401 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2461 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2521 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2581 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2641 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2701 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2761 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2821 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2881 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2941 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3001 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3061 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3121 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3181 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3241 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3301 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3361 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3421 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3481 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3541 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3601 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3661 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3721 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3781 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3841 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3901 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3961 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4021 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4081 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     4141 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     4201 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     4261 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     4321 agattgtact gagagtgcac cataaaattg taaacgttaa tattttgtta aaattcgcgt
     4381 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     4441 ataaatcaaa agaatagccc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4501 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4561 gcccactacg tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     4621 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     4681 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     4741 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     4801 actatggttg ctttgacgta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     4861 ccgcatcagg cgacgacagc gaagtgtctc gccactacaa tgctgtagta aaaatgaaaa
     4921 ggcttgttat cggcgtgcgg gatcgatccg agcttgagac aatctcttta caatacaatt
     4981 gcttacattg agcggttgtt aaatatcggc acggtaaacg atagcgaaat tacaatgctt
     5041 atagcagatt tttacgattt gtattccaat tataatattg aattgccacc accccaagcg
     5101 ctgcctcgaa gcagacgacc ttccgttgtg cagccagcgg cgcctgcgcc ggtgcccaca
     5161 atcgtgcgcg aacaaactaa accagaacaa attataccgg cggcaccgcc gccaccacct
     5221 tctcccgtgc ctaacattcc agcgcctcca ccaccaccac caccatcgat gtctgaattg
     5281 ccgcccgctc caccaatgcc gacggaacct caacccgctg cacctttaga cgacagacaa
     5341 caattgttgg aagctattag aaacgaaaaa aatcgcactc gtctcagacc ggtcaaacca
     5401 aaaacggcgc ccgaaaccag tacaatagtt gaggtgccga ctgtgttgcc taaagagaca
     5461 tttgagccta aaccgccgtc tgcatcaccg ccaccacctc cgcctccgcc tccgccgcca
     5521 gccccgcctg cgcctccacc gatggtagat ttatcatcag ctccaccacc gccgccatta
     5581 gtagatttgc cgtctgaaat gttaccaccg cctgcaccat cgctttctaa cgtgttgtct
     5641 gaattaaaat cgggcacagt tagattgaaa cccgcccaaa aacgcccgca atcagaaata
     5701 attccaaaaa gctcaactac aaatttgatc gcggacgtgt tagccgacac aattaatagg
     5761 cgtcgtgtgg ctatggcaaa atcgtcttcg gaagcaactt ctaacgacga gggttgggac
     5821 gacgacgata atcggcctaa taaagctaac acgcccgatg ttaaatatgt ccaagcttta
     5881 tttaacgtgt ttacgtcgag tcaattgtac actaacgaca gtgatgaaag aaatacaaaa
     5941 gcgcataata ttttgaacga cgtcgaacct ttattacaaa acaaaacaca aacgaatatc
     6001 gacaaagcta gattgctgct acaagatttg gcaagttttg tggcgttgag cgaaaatcca
     6061 ttagatagtc cagccatcgg ttcggaaaaa caacccttgt ttgaaactaa tcgaaaccta
     6121 ttttacaaat ctattgagga tttaatattt aaattcagat ataaagacgc tgaaaatcat
     6181 ttgattttcg ctctaacata ccaccctaaa gattataaat ttaatgaatt attaaaatac
     6241 gtacaacaat tgtctgtaaa tcaacaacgc acagaatcta gcgcttaata aatgtactaa
     6301 taacccggat caattaatcg cctaggggtt atgatagtta ttgctcagcg gtggcagcag
     6361 ccaactcagc ttcctttcgg gctttgttag cagccggatc ttctaggctc aagcagtgat
     6421 cagatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     6481 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     6541 g
//
