LOCUS       sequence-324119-        6564 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 N-ter Flag tag under the pH promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6564)
  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 6564)
  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-Flag-Nter (VE5627)
FEATURES             Location/Qualifiers
     source          1..6564
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     48..67
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_recomb     210..1567
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(226..1567)
                     /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      1714..2169
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1801..1820)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2011..2032
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2186..2205)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2305..2327
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2365..2383)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2451..2555
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2556..2624,2625..3416)
                     /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(2774..2793)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3587..4175
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4076..4095
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4329..4346
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_recomb     4738..5565
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             4738..4862
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4900..5505)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     protein_bind    complement(5572..5605)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    5795..5843
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5868..5887
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(5979..6088)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        6133..6224
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     6149..6166
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     regulatory      6266..6275
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6275..6298
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    join(6521..6564,1..91)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(join(6558..6564,1..13))
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
       61 actcatcaat gtatcttatc atgtctggat ctgatcactg cttgagccta gaagatccgg
      121 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactat
      181 cataacccct aggcgattaa ttgatccggg ttattagtac atttattaag cgctagattc
      241 tgtgcgttgt tgatttacag acaattgttg tacgtatttt aataattcat taaatttata
      301 atctttaggg tggtatgtta gagcgaaaat caaatgattt tcagcgtctt tatatctgaa
      361 tttaaatatt aaatcctcaa tagatttgta aaataggttt cgattagttt caaacaaggg
      421 ttgtttttcc gaaccgatgg ctggactatc taatggattt tcgctcaacg ccacaaaact
      481 tgccaaatct tgtagcagca atctagcttt gtcgatattc gtttgtgttt tgttttgtaa
      541 taaaggttcg acgtcgttca aaatattatg cgcttttgta tttctttcat cactgtcgtt
      601 agtgtacaat tgactcgacg taaacacgtt aaataaagct tggacatatt taacatcggg
      661 cgtgttagct ttattaggcc gattatcgtc gtcgtcccaa ccctcgtcgt tagaagttgc
      721 ttccgaagac gattttgcca tagccacacg acgcctatta attgtgtcgg ctaacacgtc
      781 cgcgatcaaa tttgtagttg agctttttgg aattatttct gattgcgggc gtttttgggc
      841 gggtttcaat ctaactgtgc ccgattttaa ttcagacaac acgttagaaa gcgatggtgc
      901 aggcggtggt aacatttcag acggcaaatc tactaatggc ggcggtggtg gagctgatga
      961 taaatctacc atcggtggag gcgcaggcgg ggctggcggc ggaggcggag gcggaggtgg
     1021 tggcggtgat gcagacggcg gtttaggctc aaatgtctct ttaggcaaca cagtcggcac
     1081 ctcaactatt gtactggttt cgggcgccgt ttttggtttg accggtctga gacgagtgcg
     1141 atttttttcg tttctaatag cttccaacaa ttgttgtctg tcgtctaaag gtgcagcggg
     1201 ttgaggttcc gtcggcattg gtggagcggg cggcaattca gacatcgatg gtggtggtgg
     1261 tggtggaggc gctggaatgt taggcacggg agaaggtggt ggcggcggtg ccgccggtat
     1321 aatttgttct ggtttagttt gttcgcgcac gattgtgggc accggcgcag gcgccgctgg
     1381 ctgcacaacg gaaggtcgtc tgcttcgagg cagcgcttgg ggtggtggca attcaatatt
     1441 ataattggaa tacaaatcgt aaaaatctgc tataagcatt gtaatttcgc tatcgtttac
     1501 cgtgccgata tttaacaacc gctcaatgta agcaattgta ttgtaaagag attgtctcaa
     1561 gctcggatcg atcccgcacg ccgataacaa gccttttcat ttttactaca gcattgtagt
     1621 ggcgagacac ttcgctgtcg tcgcctgatg cggtattttc tccttacgca tctgtgcggt
     1681 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     1741 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     1801 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     1861 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     1921 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     1981 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2041 tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2101 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     2161 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     2221 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     2281 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     2341 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     2401 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2461 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2521 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     2581 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2641 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     2701 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     2761 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     2821 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     2881 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     2941 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3001 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3061 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3121 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3181 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3241 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3301 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3361 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3421 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3481 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3541 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3601 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3661 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3721 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3781 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3841 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3901 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3961 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4021 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4081 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4141 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4201 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4261 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4321 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     4381 tccccgcgcg ttggccgatt cattaatgca gaacggctcc gcccactatt aatgaaatta
     4441 aaaattccaa ttttaaaaaa cgcagcaaga gaaacatttg tatgaaagaa tgcgtagaag
     4501 gaaagaaaaa tgtcgtcgac atgctgaaca acaagattaa tatgcctccg tgtataaaaa
     4561 aaatattgaa cgatttgaaa gaaaacaatg taccgcgcgg cggtatgtac aggaagaggt
     4621 ttatactaaa ctgttacatt gcaaacgtgg tttcgtgtgc caagtgtgaa aaccgatgtt
     4681 taatcaaggc tctgacgcat ttctacaacc acgactccaa gtgtgtgggt gaagtcatgc
     4741 atcttttaat caaatcccaa gatgtgtata aaccaccaaa ctgccaaaaa atgaaaactg
     4801 tcgacaagct ctgtccgttt gctggcaact gcaagggtct caatcctatt tgtaattatt
     4861 gaataataaa acaattataa atgtcaaatt tgttttttat taacgataca aaccaaacgc
     4921 aacaagaaca tttgtagtat tatctataat tgaaaacgcg tagttataat cgctgaggta
     4981 atatttaaaa tcattttcaa atgattcaca gttaatttgc gacaatataa ttttattttc
     5041 acataaacta gacgccttgt cgtcttcttc ttcgtattcc ttctcttttt catttttctc
     5101 ctcataaaaa ttaacatagt tattatcgta tccatatatg tatctatcgt atagagtaaa
     5161 ttttttgttg tcataaatat atatgtcttt tttaatgggg tgtatagtac cgctgcgcat
     5221 agtttttctg taatttacaa cagtgctatt ttctggtagt tcttcggagt gtgttgcttt
     5281 aattattaaa tttatataat caatgaattt gggatcgtcg gttttgtaca atatgttgcc
     5341 ggcatagtac gcagcttctt ctagttcaat tacaccattt tttagcagca ccggattaac
     5401 ataactttcc aaaatgttgt acgaaccgtt aaacaaaaac agttcacctc ccttttctat
     5461 actattgtct gcgagcagtt gtttgttgtt aaaaataaca gccattgtaa tgagacgcac
     5521 aaactaatat cacaaactgg aaatgtctat caatatatag ttgctgatat cataacttcg
     5581 tatagcatac attatacgaa gttatctggt ttaaacgtac ccgtagtggc tatggcaggg
     5641 cttgccgccc cgacgttggc tgcgagccct gggccttcac ccgaacttgg gggttggggt
     5701 ggggaaaagg aagaaacgcg ggcgtattgg tcccaatggg gtctcggtgg ggtatcgaca
     5761 gagtgccagc cctgggaccg aaccccgcgt ttatgaacaa acgacccaac acccgtgcgt
     5821 tttattctgt ctttttattg ccgtcatagc gcgggttcct tccggtattg tctccttccg
     5881 tgtttcagtt agcctccccc atctcccggt accgcatgct atgcatcagc tgctagcacc
     5941 atggctcgag atcccgggtg atcaagtctt cgtcgagtga ttgtaaataa aatgtaattt
     6001 acagtatagt attttaatta atatacaaat gatttgataa taattcttat ttaactataa
     6061 tatattgtgt tgggttgaat taaaggtccg tatactagta tcgattcgcg acctactccg
     6121 gaatattaat agatcatgga gataattaaa atgataacca tctcgcaaat aaataagtat
     6181 tttactgttt tcgtaacagt tttgtaataa aaaaacctat aaatattccg gattattcat
     6241 accgtcccac catcgggcgc ggatcgccac catggactac aaggacgacg atgacaaggg
     6301 atcccggtcc gaagcgcgcg gaattcaaag gcctacgtcg acgagctcac tagtcgcggc
     6361 cgctttcgaa tctagagcct gcagtctcga caagcttgtc gagaagtact agaggatcat
     6421 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     6481 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     6541 taatggttac aaataaagca atag
//
