LOCUS       sequence-324179-        7351 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 TwinStrep tag under the PH 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7351)
  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 7351)
  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-P10-mCherry-pH-3C-TwinStrep (VE5621)
FEATURES             Location/Qualifiers
     source          1..7351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(14..123)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        168..259
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     184..201
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             374..397
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(398..421,422..457,458..481)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="WSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     polyA_signal    630..764
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(667..686)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     721..740
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_recomb     883..2240
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(899..2240)
                     /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      2387..2842
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2474..2493)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2684..2705
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2859..2878)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2978..3000
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3038..3056)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3124..3228
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3229..3297,3298..4089)
                     /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(3447..3466)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4260..4848
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4749..4768
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5002..5019
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_recomb     5411..6238
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             5411..5535
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5573..6178)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     protein_bind    complement(6245..6278)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    6468..6516
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     6541..6560
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(6611..7321)
                     /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(6889..6908)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7158..7176
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
ORIGIN
        1 gtcttcgtcg agtgattgta aataaaatgt aatttacagt atagtatttt aattaatata
       61 caaatgattt gataataatt cttatttaac tataatatat tgtgttgggt tgaattaaag
      121 gtccgtatac tagtatcgat tcgcgaccta ctccggaata ttaatagatc atggagataa
      181 ttaaaatgat aaccatctcg caaataaata agtattttac tgttttcgta acagttttgt
      241 aataaaaaaa cctataaata ttccggatta ttcataccgt cccaccatcg ggcgcggatc
      301 ccggtccgaa gcgcgcggaa ttcaaaggcc tacgtcgacg agctcactag tcgcggccgc
      361 tttcgaatct agactggaag ttctgtttca gggtccgtgg tcacatccgc agtttgaaaa
      421 aggtggtggt agcggtggtg gttcaggtgg ttctgcttgg agccaccctc agtttgagaa
      481 atagagcctg cagtctcgac aagcttgtcg agaagtacta gaggatcata atcagccata
      541 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
      601 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
      661 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
      721 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgatca ctgcttgagc
      781 ctagaagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
      841 gagcaataac tatcataacc cctaggcgat taattgatcc gggttattag tacatttatt
      901 aagcgctaga ttctgtgcgt tgttgattta cagacaattg ttgtacgtat tttaataatt
      961 cattaaattt ataatcttta gggtggtatg ttagagcgaa aatcaaatga ttttcagcgt
     1021 ctttatatct gaatttaaat attaaatcct caatagattt gtaaaatagg tttcgattag
     1081 tttcaaacaa gggttgtttt tccgaaccga tggctggact atctaatgga ttttcgctca
     1141 acgccacaaa acttgccaaa tcttgtagca gcaatctagc tttgtcgata ttcgtttgtg
     1201 ttttgttttg taataaaggt tcgacgtcgt tcaaaatatt atgcgctttt gtatttcttt
     1261 catcactgtc gttagtgtac aattgactcg acgtaaacac gttaaataaa gcttggacat
     1321 atttaacatc gggcgtgtta gctttattag gccgattatc gtcgtcgtcc caaccctcgt
     1381 cgttagaagt tgcttccgaa gacgattttg ccatagccac acgacgccta ttaattgtgt
     1441 cggctaacac gtccgcgatc aaatttgtag ttgagctttt tggaattatt tctgattgcg
     1501 ggcgtttttg ggcgggtttc aatctaactg tgcccgattt taattcagac aacacgttag
     1561 aaagcgatgg tgcaggcggt ggtaacattt cagacggcaa atctactaat ggcggcggtg
     1621 gtggagctga tgataaatct accatcggtg gaggcgcagg cggggctggc ggcggaggcg
     1681 gaggcggagg tggtggcggt gatgcagacg gcggtttagg ctcaaatgtc tctttaggca
     1741 acacagtcgg cacctcaact attgtactgg tttcgggcgc cgtttttggt ttgaccggtc
     1801 tgagacgagt gcgatttttt tcgtttctaa tagcttccaa caattgttgt ctgtcgtcta
     1861 aaggtgcagc gggttgaggt tccgtcggca ttggtggagc gggcggcaat tcagacatcg
     1921 atggtggtgg tggtggtgga ggcgctggaa tgttaggcac gggagaaggt ggtggcggcg
     1981 gtgccgccgg tataatttgt tctggtttag tttgttcgcg cacgattgtg ggcaccggcg
     2041 caggcgccgc tggctgcaca acggaaggtc gtctgcttcg aggcagcgct tggggtggtg
     2101 gcaattcaat attataattg gaatacaaat cgtaaaaatc tgctataagc attgtaattt
     2161 cgctatcgtt taccgtgccg atatttaaca accgctcaat gtaagcaatt gtattgtaaa
     2221 gagattgtct caagctcgga tcgatcccgc acgccgataa caagcctttt catttttact
     2281 acagcattgt agtggcgaga cacttcgctg tcgtcgcctg atgcggtatt ttctccttac
     2341 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     2401 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2461 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2521 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2581 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     2641 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2701 actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg
     2761 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2821 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     2881 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     2941 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     3001 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     3061 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     3121 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3181 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     3241 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3301 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3361 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3421 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     3481 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3541 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3601 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3661 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3721 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     3781 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3841 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3901 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3961 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4021 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4081 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     4141 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4201 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4261 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4321 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4381 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4441 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4501 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4561 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4621 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4681 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4741 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4801 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4861 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4921 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4981 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     5041 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagaacggc tccgcccact
     5101 attaatgaaa ttaaaaattc caattttaaa aaacgcagca agagaaacat ttgtatgaaa
     5161 gaatgcgtag aaggaaagaa aaatgtcgtc gacatgctga acaacaagat taatatgcct
     5221 ccgtgtataa aaaaaatatt gaacgatttg aaagaaaaca atgtaccgcg cggcggtatg
     5281 tacaggaaga ggtttatact aaactgttac attgcaaacg tggtttcgtg tgccaagtgt
     5341 gaaaaccgat gtttaatcaa ggctctgacg catttctaca accacgactc caagtgtgtg
     5401 ggtgaagtca tgcatctttt aatcaaatcc caagatgtgt ataaaccacc aaactgccaa
     5461 aaaatgaaaa ctgtcgacaa gctctgtccg tttgctggca actgcaaggg tctcaatcct
     5521 atttgtaatt attgaataat aaaacaatta taaatgtcaa atttgttttt tattaacgat
     5581 acaaaccaaa cgcaacaaga acatttgtag tattatctat aattgaaaac gcgtagttat
     5641 aatcgctgag gtaatattta aaatcatttt caaatgattc acagttaatt tgcgacaata
     5701 taattttatt ttcacataaa ctagacgcct tgtcgtcttc ttcttcgtat tccttctctt
     5761 tttcattttt ctcctcataa aaattaacat agttattatc gtatccatat atgtatctat
     5821 cgtatagagt aaattttttg ttgtcataaa tatatatgtc ttttttaatg gggtgtatag
     5881 taccgctgcg catagttttt ctgtaattta caacagtgct attttctggt agttcttcgg
     5941 agtgtgttgc tttaattatt aaatttatat aatcaatgaa tttgggatcg tcggttttgt
     6001 acaatatgtt gccggcatag tacgcagctt cttctagttc aattacacca ttttttagca
     6061 gcaccggatt aacataactt tccaaaatgt tgtacgaacc gttaaacaaa aacagttcac
     6121 ctcccttttc tatactattg tctgcgagca gttgtttgtt gttaaaaata acagccattg
     6181 taatgagacg cacaaactaa tatcacaaac tggaaatgtc tatcaatata tagttgctga
     6241 tatcataact tcgtatagca tacattatac gaagttatct ggtttaaacg tacccgtagt
     6301 ggctatggca gggcttgccg ccccgacgtt ggctgcgagc cctgggcctt cacccgaact
     6361 tgggggttgg ggtggggaaa aggaagaaac gcgggcgtat tggtcccaat ggggtctcgg
     6421 tggggtatcg acagagtgcc agccctggga ccgaaccccg cgtttatgaa caaacgaccc
     6481 aacacccgtg cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta
     6541 ttgtctcctt ccgtgtttca gttagcctcc cccatctccc ggtaccgcat gctatgcatc
     6601 agctgctagc ttacttgtac agctcgtcca tgccgccggt ggagtggcgg ccctcggcgc
     6661 gttcgtactg ttccacgatg gtgtagtcct cgttgtggga ggtgatgtcc aacttgatgt
     6721 tgacgttgta ggcgccgggc agctgcacgg gcttcttggc cttgtaggtg gtcttgacct
     6781 cagcgtcgta gtggccgccg tccttcagct tcagcctctg cttgatctcg cccttcaggg
     6841 cgccgtcctc ggggtacatc cgctcggagg aggcctccca gcccatggtc ttcttctgca
     6901 ttacggggcc gtcggagggg aagttggtgc cgcgcagctt caccttgtag atgaactcgc
     6961 cgtcctgcag ggaggagtcc tgggtcacgg tcaccacgcc gccgtcctcg aagttcatca
     7021 cgcgctccca cttgaagccc tcggggaagg acagcttcaa gtagtcgggg atgtcggcgg
     7081 ggtgcttcac gtaggccttg gagccgtaca tgaactgagg ggacaggatg tcccaggcga
     7141 agggcagggg gccacccttg gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc
     7201 ggccctcgcc ctcgccctcg atctcgaact cgtggccgtt cacggagccc tccatgtgca
     7261 ccttgaagcg catgaactcc ttgatgatgg ccatgttatc ctcctcgccc ttgctcacca
     7321 tacctcctgg ctcgagatcc cgggtgatca a
//
