LOCUS       sequence-324174-        6545 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-ter6His tag under the pH promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6545)
  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 6545)
  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-6His-Cter (VE5630)
FEATURES             Location/Qualifiers
     source          1..6545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     277..1104
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             277..401
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(439..1044)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     protein_bind    complement(1111..1144)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1334..1382
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1407..1426
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(1518..1627)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        1672..1763
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1688..1705
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             1878..1895
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2041..2175
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2078..2097)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2132..2151
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_recomb     2294..3651
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(2310..3651)
                     /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      3798..4253
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3885..3904)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4095..4116
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4270..4289)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4389..4411
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4449..4467)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4535..4639
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4640..4708,4709..5500)
                     /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(4858..4877)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5671..6259
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6160..6179
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6413..6430
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcagcaagag aaacatttgt atgaaagaat gcgtagaagg aaagaaaaat gtcgtcgaca
       61 tgctgaacaa caagattaat atgcctccgt gtataaaaaa aatattgaac gatttgaaag
      121 aaaacaatgt accgcgcggc ggtatgtaca ggaagaggtt tatactaaac tgttacattg
      181 caaacgtggt ttcgtgtgcc aagtgtgaaa accgatgttt aatcaaggct ctgacgcatt
      241 tctacaacca cgactccaag tgtgtgggtg aagtcatgca tcttttaatc aaatcccaag
      301 atgtgtataa accaccaaac tgccaaaaaa tgaaaactgt cgacaagctc tgtccgtttg
      361 ctggcaactg caagggtctc aatcctattt gtaattattg aataataaaa caattataaa
      421 tgtcaaattt gttttttatt aacgatacaa accaaacgca acaagaacat ttgtagtatt
      481 atctataatt gaaaacgcgt agttataatc gctgaggtaa tatttaaaat cattttcaaa
      541 tgattcacag ttaatttgcg acaatataat tttattttca cataaactag acgccttgtc
      601 gtcttcttct tcgtattcct tctctttttc atttttctcc tcataaaaat taacatagtt
      661 attatcgtat ccatatatgt atctatcgta tagagtaaat tttttgttgt cataaatata
      721 tatgtctttt ttaatggggt gtatagtacc gctgcgcata gtttttctgt aatttacaac
      781 agtgctattt tctggtagtt cttcggagtg tgttgcttta attattaaat ttatataatc
      841 aatgaatttg ggatcgtcgg ttttgtacaa tatgttgccg gcatagtacg cagcttcttc
      901 tagttcaatt acaccatttt ttagcagcac cggattaaca taactttcca aaatgttgta
      961 cgaaccgtta aacaaaaaca gttcacctcc cttttctata ctattgtctg cgagcagttg
     1021 tttgttgtta aaaataacag ccattgtaat gagacgcaca aactaatatc acaaactgga
     1081 aatgtctatc aatatatagt tgctgatatc ataacttcgt atagcataca ttatacgaag
     1141 ttatctggtt taaacgtacc cgtagtggct atggcagggc ttgccgcccc gacgttggct
     1201 gcgagccctg ggccttcacc cgaacttggg ggttggggtg gggaaaagga agaaacgcgg
     1261 gcgtattggt cccaatgggg tctcggtggg gtatcgacag agtgccagcc ctgggaccga
     1321 accccgcgtt tatgaacaaa cgacccaaca cccgtgcgtt ttattctgtc tttttattgc
     1381 cgtcatagcg cgggttcctt ccggtattgt ctccttccgt gtttcagtta gcctccccca
     1441 tctcccggta ccgcatgcta tgcatcagct gctagcacca tggctcgaga tcccgggtga
     1501 tcaagtcttc gtcgagtgat tgtaaataaa atgtaattta cagtatagta ttttaattaa
     1561 tatacaaatg atttgataat aattcttatt taactataat atattgtgtt gggttgaatt
     1621 aaaggtccgt atactagtat cgattcgcga cctactccgg aatattaata gatcatggag
     1681 ataattaaaa tgataaccat ctcgcaaata aataagtatt ttactgtttt cgtaacagtt
     1741 ttgtaataaa aaaacctata aatattccgg attattcata ccgtcccacc atcgggcgcg
     1801 gatcccggtc cgaagcgcgc ggaattcaaa ggcctacgtc gacgagctca ctagtcgcgg
     1861 ccgctttcga atctagacac catcaccatc accattaact gcagtctcga caagcttgtc
     1921 gagaagtact agaggatcat aatcagccat accacatttg tagaggtttt acttgcttta
     1981 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     2041 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2101 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2161 tatcatgtct ggatctgatc actgcttgag cctagaagat ccggctgcta acaaagcccg
     2221 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctatcataac ccctaggcga
     2281 ttaattgatc cgggttatta gtacatttat taagcgctag attctgtgcg ttgttgattt
     2341 acagacaatt gttgtacgta ttttaataat tcattaaatt tataatcttt agggtggtat
     2401 gttagagcga aaatcaaatg attttcagcg tctttatatc tgaatttaaa tattaaatcc
     2461 tcaatagatt tgtaaaatag gtttcgatta gtttcaaaca agggttgttt ttccgaaccg
     2521 atggctggac tatctaatgg attttcgctc aacgccacaa aacttgccaa atcttgtagc
     2581 agcaatctag ctttgtcgat attcgtttgt gttttgtttt gtaataaagg ttcgacgtcg
     2641 ttcaaaatat tatgcgcttt tgtatttctt tcatcactgt cgttagtgta caattgactc
     2701 gacgtaaaca cgttaaataa agcttggaca tatttaacat cgggcgtgtt agctttatta
     2761 ggccgattat cgtcgtcgtc ccaaccctcg tcgttagaag ttgcttccga agacgatttt
     2821 gccatagcca cacgacgcct attaattgtg tcggctaaca cgtccgcgat caaatttgta
     2881 gttgagcttt ttggaattat ttctgattgc gggcgttttt gggcgggttt caatctaact
     2941 gtgcccgatt ttaattcaga caacacgtta gaaagcgatg gtgcaggcgg tggtaacatt
     3001 tcagacggca aatctactaa tggcggcggt ggtggagctg atgataaatc taccatcggt
     3061 ggaggcgcag gcggggctgg cggcggaggc ggaggcggag gtggtggcgg tgatgcagac
     3121 ggcggtttag gctcaaatgt ctctttaggc aacacagtcg gcacctcaac tattgtactg
     3181 gtttcgggcg ccgtttttgg tttgaccggt ctgagacgag tgcgattttt ttcgtttcta
     3241 atagcttcca acaattgttg tctgtcgtct aaaggtgcag cgggttgagg ttccgtcggc
     3301 attggtggag cgggcggcaa ttcagacatc gatggtggtg gtggtggtgg aggcgctgga
     3361 atgttaggca cgggagaagg tggtggcggc ggtgccgccg gtataatttg ttctggttta
     3421 gtttgttcgc gcacgattgt gggcaccggc gcaggcgccg ctggctgcac aacggaaggt
     3481 cgtctgcttc gaggcagcgc ttggggtggt ggcaattcaa tattataatt ggaatacaaa
     3541 tcgtaaaaat ctgctataag cattgtaatt tcgctatcgt ttaccgtgcc gatatttaac
     3601 aaccgctcaa tgtaagcaat tgtattgtaa agagattgtc tcaagctcgg atcgatcccg
     3661 cacgccgata acaagccttt tcatttttac tacagcattg tagtggcgag acacttcgct
     3721 gtcgtcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     3781 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3841 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3901 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3961 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     4021 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4081 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg
     4141 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4201 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     4261 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     4321 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     4381 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     4441 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     4501 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4561 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4621 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4681 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4741 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4801 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4861 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4921 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4981 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5041 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5101 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5161 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5221 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5281 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5341 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5401 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5461 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5521 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5581 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5641 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5701 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5761 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5821 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5881 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5941 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6001 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6061 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6121 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6181 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6241 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6301 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6361 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     6421 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     6481 gattcattaa tgcagaacgg ctccgcccac tattaatgaa attaaaaatt ccaattttaa
     6541 aaaac
//
