LOCUS       sequence-324116-        7887 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 mCherry cDNA under each promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7887)
  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 7887)
  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-mCh PH-mCh (VE5625)
FEATURES             Location/Qualifiers
     source          1..7887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(56..89)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    279..327
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     352..371
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(422..1132)
                     /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"
     promoter        complement(1176..1285)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        1330..1421
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1346..1363
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             1467..2177
                     /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"
     polyA_signal    2328..2462
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2365..2384)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2419..2438
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_recomb     2581..3938
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(2597..3938)
                     /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      4085..4540
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4172..4191)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4382..4403
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4557..4576)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4676..4698
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4736..4754)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4822..4926
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4927..4995,4996..5787)
                     /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(5145..5164)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5958..6546
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6447..6466
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6700..6717
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_recomb     join(7109..7887,1..49)
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             7109..7233
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(7271..7876)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
ORIGIN
        1 gcacaaacta atatcacaaa ctggaaatgt ctatcaatat atagttgctg atatcataac
       61 ttcgtatagc atacattata cgaagttatc tggtttaaac gtacccgtag tggctatggc
      121 agggcttgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttgggggttg
      181 gggtggggaa aaggaagaaa cgcgggcgta ttggtcccaa tggggtctcg gtggggtatc
      241 gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc caacacccgt
      301 gcgttttatt ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct
      361 tccgtgtttc agttagcctc ccccatctcc cggtaccgca tgctatgcat cagctgctag
      421 cttacttgta cagctcgtcc atgccgccgg tggagtggcg gccctcggcg cgttcgtact
      481 gttccacgat ggtgtagtcc tcgttgtggg aggtgatgtc caacttgatg ttgacgttgt
      541 aggcgccggg cagctgcacg ggcttcttgg ccttgtaggt ggtcttgacc tcagcgtcgt
      601 agtggccgcc gtccttcagc ttcagcctct gcttgatctc gcccttcagg gcgccgtcct
      661 cggggtacat ccgctcggag gaggcctccc agcccatggt cttcttctgc attacggggc
      721 cgtcggaggg gaagttggtg ccgcgcagct tcaccttgta gatgaactcg ccgtcctgca
      781 gggaggagtc ctgggtcacg gtcaccacgc cgccgtcctc gaagttcatc acgcgctccc
      841 acttgaagcc ctcggggaag gacagcttca agtagtcggg gatgtcggcg gggtgcttca
      901 cgtaggcctt ggagccgtac atgaactgag gggacaggat gtcccaggcg aagggcaggg
      961 ggccaccctt ggtcaccttc agcttggcgg tctgggtgcc ctcgtagggg cggccctcgc
     1021 cctcgccctc gatctcgaac tcgtggccgt tcacggagcc ctccatgtgc accttgaagc
     1081 gcatgaactc cttgatgatg gccatgttat cctcctcgcc cttgctcacc atacctcctg
     1141 gctcgagatc ccgggtgatc aagtcttcgt cgagtgattg taaataaaat gtaatttaca
     1201 gtatagtatt ttaattaata tacaaatgat ttgataataa ttcttattta actataatat
     1261 attgtgttgg gttgaattaa aggtccgtat actagtatcg attcgcgacc tactccggaa
     1321 tattaataga tcatggagat aattaaaatg ataaccatct cgcaaataaa taagtatttt
     1381 actgttttcg taacagtttt gtaataaaaa aacctataaa tattccggat tattcatacc
     1441 gtcccaccat cgggcgcgga tccaccatgg tgagcaaggg cgaggaggat aacatggcca
     1501 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
     1561 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     1621 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
     1681 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
     1741 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     1801 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     1861 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
     1921 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     1981 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     2041 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     2101 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     2161 acgagctgta caagtaatct agagcctgca gtctcgacaa gcttgtcgag aagtactaga
     2221 ggatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2281 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2341 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2401 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     2461 tctgatcact gcttgagcct agaagatccg gctgctaaca aagcccgaaa ggaagctgag
     2521 ttggctgctg ccaccgctga gcaataacta tcataacccc taggcgatta attgatccgg
     2581 gttattagta catttattaa gcgctagatt ctgtgcgttg ttgatttaca gacaattgtt
     2641 gtacgtattt taataattca ttaaatttat aatctttagg gtggtatgtt agagcgaaaa
     2701 tcaaatgatt ttcagcgtct ttatatctga atttaaatat taaatcctca atagatttgt
     2761 aaaataggtt tcgattagtt tcaaacaagg gttgtttttc cgaaccgatg gctggactat
     2821 ctaatggatt ttcgctcaac gccacaaaac ttgccaaatc ttgtagcagc aatctagctt
     2881 tgtcgatatt cgtttgtgtt ttgttttgta ataaaggttc gacgtcgttc aaaatattat
     2941 gcgcttttgt atttctttca tcactgtcgt tagtgtacaa ttgactcgac gtaaacacgt
     3001 taaataaagc ttggacatat ttaacatcgg gcgtgttagc tttattaggc cgattatcgt
     3061 cgtcgtccca accctcgtcg ttagaagttg cttccgaaga cgattttgcc atagccacac
     3121 gacgcctatt aattgtgtcg gctaacacgt ccgcgatcaa atttgtagtt gagctttttg
     3181 gaattatttc tgattgcggg cgtttttggg cgggtttcaa tctaactgtg cccgatttta
     3241 attcagacaa cacgttagaa agcgatggtg caggcggtgg taacatttca gacggcaaat
     3301 ctactaatgg cggcggtggt ggagctgatg ataaatctac catcggtgga ggcgcaggcg
     3361 gggctggcgg cggaggcgga ggcggaggtg gtggcggtga tgcagacggc ggtttaggct
     3421 caaatgtctc tttaggcaac acagtcggca cctcaactat tgtactggtt tcgggcgccg
     3481 tttttggttt gaccggtctg agacgagtgc gatttttttc gtttctaata gcttccaaca
     3541 attgttgtct gtcgtctaaa ggtgcagcgg gttgaggttc cgtcggcatt ggtggagcgg
     3601 gcggcaattc agacatcgat ggtggtggtg gtggtggagg cgctggaatg ttaggcacgg
     3661 gagaaggtgg tggcggcggt gccgccggta taatttgttc tggtttagtt tgttcgcgca
     3721 cgattgtggg caccggcgca ggcgccgctg gctgcacaac ggaaggtcgt ctgcttcgag
     3781 gcagcgcttg gggtggtggc aattcaatat tataattgga atacaaatcg taaaaatctg
     3841 ctataagcat tgtaatttcg ctatcgttta ccgtgccgat atttaacaac cgctcaatgt
     3901 aagcaattgt attgtaaaga gattgtctca agctcggatc gatcccgcac gccgataaca
     3961 agccttttca tttttactac agcattgtag tggcgagaca cttcgctgtc gtcgcctgat
     4021 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     4081 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4141 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4201 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4261 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     4321 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4381 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat
     4441 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4501 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     4561 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     4621 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4681 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     4741 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     4801 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4861 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4921 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4981 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5041 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5101 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5161 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5221 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5281 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5341 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5401 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5461 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5521 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5581 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5641 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5701 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5761 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5821 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5881 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5941 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6001 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6061 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6121 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6181 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6241 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6301 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6361 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6421 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6481 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6541 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6601 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6661 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6721 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     6781 agaacggctc cgcccactat taatgaaatt aaaaattcca attttaaaaa acgcagcaag
     6841 agaaacattt gtatgaaaga atgcgtagaa ggaaagaaaa atgtcgtcga catgctgaac
     6901 aacaagatta atatgcctcc gtgtataaaa aaaatattga acgatttgaa agaaaacaat
     6961 gtaccgcgcg gcggtatgta caggaagagg tttatactaa actgttacat tgcaaacgtg
     7021 gtttcgtgtg ccaagtgtga aaaccgatgt ttaatcaagg ctctgacgca tttctacaac
     7081 cacgactcca agtgtgtggg tgaagtcatg catcttttaa tcaaatccca agatgtgtat
     7141 aaaccaccaa actgccaaaa aatgaaaact gtcgacaagc tctgtccgtt tgctggcaac
     7201 tgcaagggtc tcaatcctat ttgtaattat tgaataataa aacaattata aatgtcaaat
     7261 ttgtttttta ttaacgatac aaaccaaacg caacaagaac atttgtagta ttatctataa
     7321 ttgaaaacgc gtagttataa tcgctgaggt aatatttaaa atcattttca aatgattcac
     7381 agttaatttg cgacaatata attttatttt cacataaact agacgccttg tcgtcttctt
     7441 cttcgtattc cttctctttt tcatttttct cctcataaaa attaacatag ttattatcgt
     7501 atccatatat gtatctatcg tatagagtaa attttttgtt gtcataaata tatatgtctt
     7561 ttttaatggg gtgtatagta ccgctgcgca tagtttttct gtaatttaca acagtgctat
     7621 tttctggtag ttcttcggag tgtgttgctt taattattaa atttatataa tcaatgaatt
     7681 tgggatcgtc ggttttgtac aatatgttgc cggcatagta cgcagcttct tctagttcaa
     7741 ttacaccatt ttttagcagc accggattaa cataactttc caaaatgttg tacgaaccgt
     7801 taaacaaaaa cagttcacct cccttttcta tactattgtc tgcgagcagt tgtttgttgt
     7861 taaaaataac agccattgta atgagac
//
