LOCUS       sequence-324127-        7237 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 N-ter HA tag under the pH promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7237)
  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 7237)
  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-HA-Nter-P10-mCherry (VE5740)
FEATURES             Location/Qualifiers
     source          1..7237
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    18..66
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     91..110
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(161..871)
                     /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(439..458)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     708..726
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     promoter        complement(915..1024)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        1069..1160
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1085..1102
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             1213..1239
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      1247..1256
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1417..1551
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1454..1473)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1508..1527
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_recomb     1670..3027
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(1686..3027)
                     /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      3174..3629
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3261..3280)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3471..3492
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3646..3665)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3765..3787
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3825..3843)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3911..4015
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4016..4084,4085..4876)
                     /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(4234..4253)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5047..5635
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5536..5555
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5789..5806
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_recomb     6198..7025
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             6198..6322
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(6360..6965)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     protein_bind    complement(7032..7065)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 ccgaaccccg cgtttatgaa caaacgaccc aacacccgtg cgttttattc tgtcttttta
       61 ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc
      121 cccatctccc ggtaccgcat gctatgcatc agctgctagc ttacttgtac agctcgtcca
      181 tgccgccggt ggagtggcgg ccctcggcgc gttcgtactg ttccacgatg gtgtagtcct
      241 cgttgtggga ggtgatgtcc aacttgatgt tgacgttgta ggcgccgggc agctgcacgg
      301 gcttcttggc cttgtaggtg gtcttgacct cagcgtcgta gtggccgccg tccttcagct
      361 tcagcctctg cttgatctcg cccttcaggg cgccgtcctc ggggtacatc cgctcggagg
      421 aggcctccca gcccatggtc ttcttctgca ttacggggcc gtcggagggg aagttggtgc
      481 cgcgcagctt caccttgtag atgaactcgc cgtcctgcag ggaggagtcc tgggtcacgg
      541 tcaccacgcc gccgtcctcg aagttcatca cgcgctccca cttgaagccc tcggggaagg
      601 acagcttcaa gtagtcgggg atgtcggcgg ggtgcttcac gtaggccttg gagccgtaca
      661 tgaactgagg ggacaggatg tcccaggcga agggcagggg gccacccttg gtcaccttca
      721 gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact
      781 cgtggccgtt cacggagccc tccatgtgca ccttgaagcg catgaactcc ttgatgatgg
      841 ccatgttatc ctcctcgccc ttgctcacca tacctcctgg ctcgagatcc cgggtgatca
      901 agtcttcgtc gagtgattgt aaataaaatg taatttacag tatagtattt taattaatat
      961 acaaatgatt tgataataat tcttatttaa ctataatata ttgtgttggg ttgaattaaa
     1021 ggtccgtata ctagtatcga ttcgcgacct actccggaat attaatagat catggagata
     1081 attaaaatga taaccatctc gcaaataaat aagtatttta ctgttttcgt aacagttttg
     1141 taataaaaaa acctataaat attccggatt attcataccg tcccaccatc gggcgcggat
     1201 cccggtccga tgtacccata cgatgttcca gattacgctc ttatggccat ggcggccgct
     1261 ttcgaatcta gagcctgcag tctcgacaag cttgtcgaga agtactagag gatcataatc
     1321 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     1381 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     1441 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1501 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ctgatcactg
     1561 cttgagccta gaagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     1621 caccgctgag caataactat cataacccct aggcgattaa ttgatccggg ttattagtac
     1681 atttattaag cgctagattc tgtgcgttgt tgatttacag acaattgttg tacgtatttt
     1741 aataattcat taaatttata atctttaggg tggtatgtta gagcgaaaat caaatgattt
     1801 tcagcgtctt tatatctgaa tttaaatatt aaatcctcaa tagatttgta aaataggttt
     1861 cgattagttt caaacaaggg ttgtttttcc gaaccgatgg ctggactatc taatggattt
     1921 tcgctcaacg ccacaaaact tgccaaatct tgtagcagca atctagcttt gtcgatattc
     1981 gtttgtgttt tgttttgtaa taaaggttcg acgtcgttca aaatattatg cgcttttgta
     2041 tttctttcat cactgtcgtt agtgtacaat tgactcgacg taaacacgtt aaataaagct
     2101 tggacatatt taacatcggg cgtgttagct ttattaggcc gattatcgtc gtcgtcccaa
     2161 ccctcgtcgt tagaagttgc ttccgaagac gattttgcca tagccacacg acgcctatta
     2221 attgtgtcgg ctaacacgtc cgcgatcaaa tttgtagttg agctttttgg aattatttct
     2281 gattgcgggc gtttttgggc gggtttcaat ctaactgtgc ccgattttaa ttcagacaac
     2341 acgttagaaa gcgatggtgc aggcggtggt aacatttcag acggcaaatc tactaatggc
     2401 ggcggtggtg gagctgatga taaatctacc atcggtggag gcgcaggcgg ggctggcggc
     2461 ggaggcggag gcggaggtgg tggcggtgat gcagacggcg gtttaggctc aaatgtctct
     2521 ttaggcaaca cagtcggcac ctcaactatt gtactggttt cgggcgccgt ttttggtttg
     2581 accggtctga gacgagtgcg atttttttcg tttctaatag cttccaacaa ttgttgtctg
     2641 tcgtctaaag gtgcagcggg ttgaggttcc gtcggcattg gtggagcggg cggcaattca
     2701 gacatcgatg gtggtggtgg tggtggaggc gctggaatgt taggcacggg agaaggtggt
     2761 ggcggcggtg ccgccggtat aatttgttct ggtttagttt gttcgcgcac gattgtgggc
     2821 accggcgcag gcgccgctgg ctgcacaacg gaaggtcgtc tgcttcgagg cagcgcttgg
     2881 ggtggtggca attcaatatt ataattggaa tacaaatcgt aaaaatctgc tataagcatt
     2941 gtaatttcgc tatcgtttac cgtgccgata tttaacaacc gctcaatgta agcaattgta
     3001 ttgtaaagag attgtctcaa gctcggatcg atcccgcacg ccgataacaa gccttttcat
     3061 ttttactaca gcattgtagt ggcgagacac ttcgctgtcg tcgcctgatg cggtattttc
     3121 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     3181 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3241 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3301 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3361 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     3421 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3481 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
     3541 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3601 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     3661 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3721 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3781 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     3841 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3901 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3961 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4021 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4081 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4141 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4201 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4261 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4321 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4381 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4441 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4501 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4561 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4621 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4681 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4741 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4801 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4861 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4921 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4981 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5041 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5101 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5161 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5221 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5281 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5341 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5401 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5461 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5521 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5581 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5641 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5701 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     5761 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     5821 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gaacggctcc
     5881 gcccactatt aatgaaatta aaaattccaa ttttaaaaaa cgcagcaaga gaaacatttg
     5941 tatgaaagaa tgcgtagaag gaaagaaaaa tgtcgtcgac atgctgaaca acaagattaa
     6001 tatgcctccg tgtataaaaa aaatattgaa cgatttgaaa gaaaacaatg taccgcgcgg
     6061 cggtatgtac aggaagaggt ttatactaaa ctgttacatt gcaaacgtgg tttcgtgtgc
     6121 caagtgtgaa aaccgatgtt taatcaaggc tctgacgcat ttctacaacc acgactccaa
     6181 gtgtgtgggt gaagtcatgc atcttttaat caaatcccaa gatgtgtata aaccaccaaa
     6241 ctgccaaaaa atgaaaactg tcgacaagct ctgtccgttt gctggcaact gcaagggtct
     6301 caatcctatt tgtaattatt gaataataaa acaattataa atgtcaaatt tgttttttat
     6361 taacgataca aaccaaacgc aacaagaaca tttgtagtat tatctataat tgaaaacgcg
     6421 tagttataat cgctgaggta atatttaaaa tcattttcaa atgattcaca gttaatttgc
     6481 gacaatataa ttttattttc acataaacta gacgccttgt cgtcttcttc ttcgtattcc
     6541 ttctcttttt catttttctc ctcataaaaa ttaacatagt tattatcgta tccatatatg
     6601 tatctatcgt atagagtaaa ttttttgttg tcataaatat atatgtcttt tttaatgggg
     6661 tgtatagtac cgctgcgcat agtttttctg taatttacaa cagtgctatt ttctggtagt
     6721 tcttcggagt gtgttgcttt aattattaaa tttatataat caatgaattt gggatcgtcg
     6781 gttttgtaca atatgttgcc ggcatagtac gcagcttctt ctagttcaat tacaccattt
     6841 tttagcagca ccggattaac ataactttcc aaaatgttgt acgaaccgtt aaacaaaaac
     6901 agttcacctc ccttttctat actattgtct gcgagcagtt gtttgttgtt aaaaataaca
     6961 gccattgtaa tgagacgcac aaactaatat cacaaactgg aaatgtctat caatatatag
     7021 ttgctgatat cataacttcg tatagcatac attatacgaa gttatctggt ttaaacgtac
     7081 ccgtagtggc tatggcaggg cttgccgccc cgacgttggc tgcgagccct gggccttcac
     7141 ccgaacttgg gggttggggt ggggaaaagg aagaaacgcg ggcgtattgg tcccaatggg
     7201 gtctcggtgg ggtatcgaca gagtgccagc cctggga
//
