LOCUS       sequence-317608-        6897 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 p10 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6897)
  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 6897)
  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-10His-Cter (VE5631)
FEATURES             Location/Qualifiers
     source          1..6897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1..34)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    224..272
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     297..316
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(346..375)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     promoter        complement(423..532)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        577..668
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     593..610
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    927..1061
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(964..983)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1018..1037
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_recomb     1180..2537
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(1196..2537)
                     /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      2684..3139
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2771..2790)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2981..3002
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3156..3175)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3275..3297
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3335..3353)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3421..3525
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3526..3594,3595..4386)
                     /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(3744..3763)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4557..5145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5046..5065
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5299..5316
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_recomb     5708..6535
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             5708..5832
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5870..6475)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
ORIGIN
        1 ataacttcgt atagcataca ttatacgaag ttatctggtt taaacgtacc cgtagtggct
       61 atggcagggc ttgccgcccc gacgttggct gcgagccctg ggccttcacc cgaacttggg
      121 ggttggggtg gggaaaagga agaaacgcgg gcgtattggt cccaatgggg tctcggtggg
      181 gtatcgacag agtgccagcc ctgggaccga accccgcgtt tatgaacaaa cgacccaaca
      241 cccgtgcgtt ttattctgtc tttttattgc cgtcatagcg cgggttcctt ccggtattgt
      301 ctccttccgt gtttcagtta gcctccccca tctcccggta ccttaatggt ggtgatggtg
      361 atggtgatga tggtggctag caccatggct cgagatcccg ggtgatcaag tcttcgtcga
      421 gtgattgtaa ataaaatgta atttacagta tagtatttta attaatatac aaatgatttg
      481 ataataattc ttatttaact ataatatatt gtgttgggtt gaattaaagg tccgtatact
      541 agtatcgatt cgcgacctac tccggaatat taatagatca tggagataat taaaatgata
      601 accatctcgc aaataaataa gtattttact gttttcgtaa cagttttgta ataaaaaaac
      661 ctataaatat tccggattat tcataccgtc ccaccatcgg gcgcggatcc cggtccgaag
      721 cgcgcggaat tcaaaggcct acgtcgacga gctcactagt cgcggccgct ttcgaatcta
      781 gagcctgcag tctcgacaag cttgtcgaga agtactagag gatcataatc agccatacca
      841 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
      901 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
      961 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1021 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctgatcactg cttgagccta
     1081 gaagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     1141 caataactat cataacccct aggcgattaa ttgatccggg ttattagtac atttattaag
     1201 cgctagattc tgtgcgttgt tgatttacag acaattgttg tacgtatttt aataattcat
     1261 taaatttata atctttaggg tggtatgtta gagcgaaaat caaatgattt tcagcgtctt
     1321 tatatctgaa tttaaatatt aaatcctcaa tagatttgta aaataggttt cgattagttt
     1381 caaacaaggg ttgtttttcc gaaccgatgg ctggactatc taatggattt tcgctcaacg
     1441 ccacaaaact tgccaaatct tgtagcagca atctagcttt gtcgatattc gtttgtgttt
     1501 tgttttgtaa taaaggttcg acgtcgttca aaatattatg cgcttttgta tttctttcat
     1561 cactgtcgtt agtgtacaat tgactcgacg taaacacgtt aaataaagct tggacatatt
     1621 taacatcggg cgtgttagct ttattaggcc gattatcgtc gtcgtcccaa ccctcgtcgt
     1681 tagaagttgc ttccgaagac gattttgcca tagccacacg acgcctatta attgtgtcgg
     1741 ctaacacgtc cgcgatcaaa tttgtagttg agctttttgg aattatttct gattgcgggc
     1801 gtttttgggc gggtttcaat ctaactgtgc ccgattttaa ttcagacaac acgttagaaa
     1861 gcgatggtgc aggcggtggt aacatttcag acggcaaatc tactaatggc ggcggtggtg
     1921 gagctgatga taaatctacc atcggtggag gcgcaggcgg ggctggcggc ggaggcggag
     1981 gcggaggtgg tggcggtgat gcagacggcg gtttaggctc aaatgtctct ttaggcaaca
     2041 cagtcggcac ctcaactatt gtactggttt cgggcgccgt ttttggtttg accggtctga
     2101 gacgagtgcg atttttttcg tttctaatag cttccaacaa ttgttgtctg tcgtctaaag
     2161 gtgcagcggg ttgaggttcc gtcggcattg gtggagcggg cggcaattca gacatcgatg
     2221 gtggtggtgg tggtggaggc gctggaatgt taggcacggg agaaggtggt ggcggcggtg
     2281 ccgccggtat aatttgttct ggtttagttt gttcgcgcac gattgtgggc accggcgcag
     2341 gcgccgctgg ctgcacaacg gaaggtcgtc tgcttcgagg cagcgcttgg ggtggtggca
     2401 attcaatatt ataattggaa tacaaatcgt aaaaatctgc tataagcatt gtaatttcgc
     2461 tatcgtttac cgtgccgata tttaacaacc gctcaatgta agcaattgta ttgtaaagag
     2521 attgtctcaa gctcggatcg atcccgcacg ccgataacaa gccttttcat ttttactaca
     2581 gcattgtagt ggcgagacac ttcgctgtcg tcgcctgatg cggtattttc tccttacgca
     2641 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     2701 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2761 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2821 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2881 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2941 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3001 ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     3061 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     3121 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     3181 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     3241 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     3301 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     3361 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     3421 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3481 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3541 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3601 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3661 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3721 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     3781 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3841 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3901 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3961 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4021 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4081 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4141 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4201 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4261 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4321 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     4381 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     4441 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4501 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4561 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4621 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4681 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     4741 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4801 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4861 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4921 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4981 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5041 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5101 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5161 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5221 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     5281 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     5341 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gaacggctcc gcccactatt
     5401 aatgaaatta aaaattccaa ttttaaaaaa cgcagcaaga gaaacatttg tatgaaagaa
     5461 tgcgtagaag gaaagaaaaa tgtcgtcgac atgctgaaca acaagattaa tatgcctccg
     5521 tgtataaaaa aaatattgaa cgatttgaaa gaaaacaatg taccgcgcgg cggtatgtac
     5581 aggaagaggt ttatactaaa ctgttacatt gcaaacgtgg tttcgtgtgc caagtgtgaa
     5641 aaccgatgtt taatcaaggc tctgacgcat ttctacaacc acgactccaa gtgtgtgggt
     5701 gaagtcatgc atcttttaat caaatcccaa gatgtgtata aaccaccaaa ctgccaaaaa
     5761 atgaaaactg tcgacaagct ctgtccgttt gctggcaact gcaagggtct caatcctatt
     5821 tgtaattatt gaataataaa acaattataa atgctaaatt tgttttttat taacgataca
     5881 aaccaaacgc aacaagaaca tttgtagtat tatctataat tgaaaacgcg tagttataat
     5941 cgctgaggta atatttaaaa tcattttcaa atgattcaca gttaatttgc gacaatataa
     6001 ttttattttc acataaacta gacgccttgt cgtcttcttc ttcgtattcc ttctcttttt
     6061 catttttctc ctcataaaaa ttaacatagt tattatcgta tccatatatg tatctatcgt
     6121 atagagtaaa ttttttgttg tcataaatat atatgtcttt tttaatgggg tgtatagtac
     6181 cgctgcgcat agtttttctg taatttacaa cagtgctatt ttctggtagt tcttcggagt
     6241 gtgttgcttt aattattaaa tttatataat caatgaattt gggatcgtcg gttttgtaca
     6301 atatgttgcc ggcatagtac gcagcttctt ctagttcaat tacaccattt tttagcagca
     6361 ccggattaac ataactttcc aaaatgttgt acgaaccgtt aaacaaaaac agttcacctc
     6421 ccttttctat actattgtct gcgagcagtt gtttgttgtt aaaaataaca gccattgtaa
     6481 tgagacgcac aaactaatat cacaaactgg aaatgtctat caatatatag ttgctgatat
     6541 cccattttac caccataata cttaaaatga tccacaaaca agccaccacc aatcgctaca
     6601 ctcaccgccg ctacagacgc tacaatcacc tcatatttat tctcctcaca tatttattct
     6661 attctaattc tcaatcatac caatcaactt ctcatactca atcataccaa tcaatacaca
     6721 ttatactact actcatttcc aatcattcac attcataata tatctgttat actactcctt
     6781 atcactactc tctccatcca cattattatc ctccatcata cgacgctaca taactacgtg
     6841 gttcctaact acaccaagca tagccctagc cagccagcca caccttacag ccttttt
//
