LOCUS       sequence-324177-        7239 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Transfer vector for multigene expression to generate recombinant 
            baculoviruses by homologous recombination. Contains a p10/pH dual 
            expression cassette with mCherry cDNA under p10 to monitor 
            infection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7239)
  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 7239)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-mCh (VE5626)
FEATURES             Location/Qualifiers
     source          1..7239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     113..130
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_recomb     522..1349
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             522..646
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(684..1289)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     protein_bind    complement(1356..1389)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1579..1627
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1652..1671
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1722..2432)
                     /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(2000..2019)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2269..2287
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     promoter        complement(2476..2585)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        2630..2721
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2646..2663
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    2980..3114
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3017..3036)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3071..3090
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_recomb     3233..4590
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(3249..4590)
                     /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      4737..5192
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4824..4843)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5034..5055
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5209..5228)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5328..5350
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5388..5406)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5474..5578
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5579..5647,5648..6439)
                     /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(5797..5816)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6610..7198
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7099..7118
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
       61 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
      121 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
      181 gattcattaa tgcagaacgg ctccgcccac tattaatgaa attaaaaatt ccaattttaa
      241 aaaacgcagc aagagaaaca tttgtatgaa agaatgcgta gaaggaaaga aaaatgtcgt
      301 cgacatgctg aacaacaaga ttaatatgcc tccgtgtata aaaaaaatat tgaacgattt
      361 gaaagaaaac aatgtaccgc gcggcggtat gtacaggaag aggtttatac taaactgtta
      421 cattgcaaac gtggtttcgt gtgccaagtg tgaaaaccga tgtttaatca aggctctgac
      481 gcatttctac aaccacgact ccaagtgtgt gggtgaagtc atgcatcttt taatcaaatc
      541 ccaagatgtg tataaaccac caaactgcca aaaaatgaaa actgtcgaca agctctgtcc
      601 gtttgctggc aactgcaagg gtctcaatcc tatttgtaat tattgaataa taaaacaatt
      661 ataaatgtca aatttgtttt ttattaacga tacaaaccaa acgcaacaag aacatttgta
      721 gtattatcta taattgaaaa cgcgtagtta taatcgctga ggtaatattt aaaatcattt
      781 tcaaatgatt cacagttaat ttgcgacaat ataattttat tttcacataa actagacgcc
      841 ttgtcgtctt cttcttcgta ttccttctct ttttcatttt tctcctcata aaaattaaca
      901 tagttattat cgtatccata tatgtatcta tcgtatagag taaatttttt gttgtcataa
      961 atatatatgt cttttttaat ggggtgtata gtaccgctgc gcatagtttt tctgtaattt
     1021 acaacagtgc tattttctgg tagttcttcg gagtgtgttg ctttaattat taaatttata
     1081 taatcaatga atttgggatc gtcggttttg tacaatatgt tgccggcata gtacgcagct
     1141 tcttctagtt caattacacc attttttagc agcaccggat taacataact ttccaaaatg
     1201 ttgtacgaac cgttaaacaa aaacagttca cctccctttt ctatactatt gtctgcgagc
     1261 agttgtttgt tgttaaaaat aacagccatt gtaatgagac gcacaaacta atatcacaaa
     1321 ctggaaatgt ctatcaatat atagttgctg atatcataac ttcgtatagc atacattata
     1381 cgaagttatc tggtttaaac gtacccgtag tggctatggc agggcttgcc gccccgacgt
     1441 tggctgcgag ccctgggcct tcacccgaac ttgggggttg gggtggggaa aaggaagaaa
     1501 cgcgggcgta ttggtcccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg
     1561 accgaacccc gcgtttatga acaaacgacc caacacccgt gcgttttatt ctgtcttttt
     1621 attgccgtca tagcgcgggt tccttccggt attgtctcct tccgtgtttc agttagcctc
     1681 ccccatctcc cggtaccgca tgctatgcat cagctgctag cttacttgta cagctcgtcc
     1741 atgccgccgg tggagtggcg gccctcggcg cgttcgtact gttccacgat ggtgtagtcc
     1801 tcgttgtggg aggtgatgtc caacttgatg ttgacgttgt aggcgccggg cagctgcacg
     1861 ggcttcttgg ccttgtaggt ggtcttgacc tcagcgtcgt agtggccgcc gtccttcagc
     1921 ttcagcctct gcttgatctc gcccttcagg gcgccgtcct cggggtacat ccgctcggag
     1981 gaggcctccc agcccatggt cttcttctgc attacggggc cgtcggaggg gaagttggtg
     2041 ccgcgcagct tcaccttgta gatgaactcg ccgtcctgca gggaggagtc ctgggtcacg
     2101 gtcaccacgc cgccgtcctc gaagttcatc acgcgctccc acttgaagcc ctcggggaag
     2161 gacagcttca agtagtcggg gatgtcggcg gggtgcttca cgtaggcctt ggagccgtac
     2221 atgaactgag gggacaggat gtcccaggcg aagggcaggg ggccaccctt ggtcaccttc
     2281 agcttggcgg tctgggtgcc ctcgtagggg cggccctcgc cctcgccctc gatctcgaac
     2341 tcgtggccgt tcacggagcc ctccatgtgc accttgaagc gcatgaactc cttgatgatg
     2401 gccatgttat cctcctcgcc cttgctcacc atacctcctg gctcgagatc ccgggtgatc
     2461 aagtcttcgt cgagtgattg taaataaaat gtaatttaca gtatagtatt ttaattaata
     2521 tacaaatgat ttgataataa ttcttattta actataatat attgtgttgg gttgaattaa
     2581 aggtccgtat actagtatcg attcgcgacc tactccggaa tattaataga tcatggagat
     2641 aattaaaatg ataaccatct cgcaaataaa taagtatttt actgttttcg taacagtttt
     2701 gtaataaaaa aacctataaa tattccggat tattcatacc gtcccaccat cgggcgcgga
     2761 tcccggtccg aagcgcgcgg aattcaaagg cctacgtcga cgagctcact agtcgcggcc
     2821 gctttcgaat ctagagcctg cagtctcgac aagcttgtcg agaagtacta gaggatcata
     2881 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2941 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     3001 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3061 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgatca
     3121 ctgcttgagc ctagaagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     3181 tgccaccgct gagcaataac tatcataacc cctaggcgat taattgatcc gggttattag
     3241 tacatttatt aagcgctaga ttctgtgcgt tgttgattta cagacaattg ttgtacgtat
     3301 tttaataatt cattaaattt ataatcttta gggtggtatg ttagagcgaa aatcaaatga
     3361 ttttcagcgt ctttatatct gaatttaaat attaaatcct caatagattt gtaaaatagg
     3421 tttcgattag tttcaaacaa gggttgtttt tccgaaccga tggctggact atctaatgga
     3481 ttttcgctca acgccacaaa acttgccaaa tcttgtagca gcaatctagc tttgtcgata
     3541 ttcgtttgtg ttttgttttg taataaaggt tcgacgtcgt tcaaaatatt atgcgctttt
     3601 gtatttcttt catcactgtc gttagtgtac aattgactcg acgtaaacac gttaaataaa
     3661 gcttggacat atttaacatc gggcgtgtta gctttattag gccgattatc gtcgtcgtcc
     3721 caaccctcgt cgttagaagt tgcttccgaa gacgattttg ccatagccac acgacgccta
     3781 ttaattgtgt cggctaacac gtccgcgatc aaatttgtag ttgagctttt tggaattatt
     3841 tctgattgcg ggcgtttttg ggcgggtttc aatctaactg tgcccgattt taattcagac
     3901 aacacgttag aaagcgatgg tgcaggcggt ggtaacattt cagacggcaa atctactaat
     3961 ggcggcggtg gtggagctga tgataaatct accatcggtg gaggcgcagg cggggctggc
     4021 ggcggaggcg gaggcggagg tggtggcggt gatgcagacg gcggtttagg ctcaaatgtc
     4081 tctttaggca acacagtcgg cacctcaact attgtactgg tttcgggcgc cgtttttggt
     4141 ttgaccggtc tgagacgagt gcgatttttt tcgtttctaa tagcttccaa caattgttgt
     4201 ctgtcgtcta aaggtgcagc gggttgaggt tccgtcggca ttggtggagc gggcggcaat
     4261 tcagacatcg atggtggtgg tggtggtgga ggcgctggaa tgttaggcac gggagaaggt
     4321 ggtggcggcg gtgccgccgg tataatttgt tctggtttag tttgttcgcg cacgattgtg
     4381 ggcaccggcg caggcgccgc tggctgcaca acggaaggtc gtctgcttcg aggcagcgct
     4441 tggggtggtg gcaattcaat attataattg gaatacaaat cgtaaaaatc tgctataagc
     4501 attgtaattt cgctatcgtt taccgtgccg atatttaaca accgctcaat gtaagcaatt
     4561 gtattgtaaa gagattgtct caagctcgga tcgatcccgc acgccgataa caagcctttt
     4621 catttttact acagcattgt agtggcgaga cacttcgctg tcgtcgcctg atgcggtatt
     4681 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     4741 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4801 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4861 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4921 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     4981 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     5041 cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg
     5101 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5161 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     5221 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     5281 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     5341 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     5401 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     5461 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5521 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5581 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5641 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     5701 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5761 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5821 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5881 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5941 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6001 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6061 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6121 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6181 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6241 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6301 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6361 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6421 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6481 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6541 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6601 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6661 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6721 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6781 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6841 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6901 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6961 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     7021 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7081 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7141 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7201 cgccagcaac gcggcctttt tacggttcct ggccttttg
//
