LOCUS       sequence-324156-        6554 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 HA tag under the pH promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6554)
  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 6554)
  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-Cter (VE5629)
FEATURES             Location/Qualifiers
     source          1..6554
                     /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..1957
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2103..2237
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2140..2159)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2194..2213
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_recomb     2356..3713
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(2372..3713)
                     /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      3860..4315
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3947..3966)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4157..4178
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4332..4351)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4451..4473
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4511..4529)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4597..4701
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4702..4770,4771..5562)
                     /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(4920..4939)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5733..6321
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6222..6241
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6475..6492
                     /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 tacccatacg atgttccaga ttacgcttaa ctgcagtctc gacaagcttg
     1981 tcgagaagta ctagaggatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     2041 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     2101 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     2161 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     2221 cttatcatgt ctggatctga tcactgcttg agcctagaag atccggctgc taacaaagcc
     2281 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactatcata acccctaggc
     2341 gattaattga tccgggttat tagtacattt attaagcgct agattctgtg cgttgttgat
     2401 ttacagacaa ttgttgtacg tattttaata attcattaaa tttataatct ttagggtggt
     2461 atgttagagc gaaaatcaaa tgattttcag cgtctttata tctgaattta aatattaaat
     2521 cctcaataga tttgtaaaat aggtttcgat tagtttcaaa caagggttgt ttttccgaac
     2581 cgatggctgg actatctaat ggattttcgc tcaacgccac aaaacttgcc aaatcttgta
     2641 gcagcaatct agctttgtcg atattcgttt gtgttttgtt ttgtaataaa ggttcgacgt
     2701 cgttcaaaat attatgcgct tttgtatttc tttcatcact gtcgttagtg tacaattgac
     2761 tcgacgtaaa cacgttaaat aaagcttgga catatttaac atcgggcgtg ttagctttat
     2821 taggccgatt atcgtcgtcg tcccaaccct cgtcgttaga agttgcttcc gaagacgatt
     2881 ttgccatagc cacacgacgc ctattaattg tgtcggctaa cacgtccgcg atcaaatttg
     2941 tagttgagct ttttggaatt atttctgatt gcgggcgttt ttgggcgggt ttcaatctaa
     3001 ctgtgcccga ttttaattca gacaacacgt tagaaagcga tggtgcaggc ggtggtaaca
     3061 tttcagacgg caaatctact aatggcggcg gtggtggagc tgatgataaa tctaccatcg
     3121 gtggaggcgc aggcggggct ggcggcggag gcggaggcgg aggtggtggc ggtgatgcag
     3181 acggcggttt aggctcaaat gtctctttag gcaacacagt cggcacctca actattgtac
     3241 tggtttcggg cgccgttttt ggtttgaccg gtctgagacg agtgcgattt ttttcgtttc
     3301 taatagcttc caacaattgt tgtctgtcgt ctaaaggtgc agcgggttga ggttccgtcg
     3361 gcattggtgg agcgggcggc aattcagaca tcgatggtgg tggtggtggt ggaggcgctg
     3421 gaatgttagg cacgggagaa ggtggtggcg gcggtgccgc cggtataatt tgttctggtt
     3481 tagtttgttc gcgcacgatt gtgggcaccg gcgcaggcgc cgctggctgc acaacggaag
     3541 gtcgtctgct tcgaggcagc gcttggggtg gtggcaattc aatattataa ttggaataca
     3601 aatcgtaaaa atctgctata agcattgtaa tttcgctatc gtttaccgtg ccgatattta
     3661 acaaccgctc aatgtaagca attgtattgt aaagagattg tctcaagctc ggatcgatcc
     3721 cgcacgccga taacaagcct tttcattttt actacagcat tgtagtggcg agacacttcg
     3781 ctgtcgtcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3841 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3901 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3961 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4021 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     4081 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4141 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4201 ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4261 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     4321 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4381 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4441 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4501 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4561 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4621 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4681 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4741 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4801 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4861 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4921 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4981 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5041 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5101 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5161 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5221 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5281 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5341 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5401 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5461 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5521 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5581 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5641 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5701 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5761 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5821 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5881 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5941 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6001 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6061 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6121 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6181 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6241 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6301 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6361 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6421 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     6481 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     6541 ccgattcatt aatg
//
