LOCUS       sequence-324153-        6551 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-ter Flag tag under the pH promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6551)
  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 6551)
  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-Flag-Cter (VE5739)
FEATURES             Location/Qualifiers
     source          1..6551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     108..125
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_recomb     517..1344
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             517..641
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(679..1284)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     protein_bind    complement(1351..1384)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1574..1622
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1647..1666
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(1758..1867)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        1912..2003
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1928..1945
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2118..2141
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2287..2421
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2324..2343)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2378..2397
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_recomb     2540..3897
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(2556..3897)
                     /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      4044..4499
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4131..4150)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4341..4362
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4516..4535)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4635..4657
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4695..4713)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4781..4885
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4886..4954,4955..5746)
                     /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(5104..5123)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5917..6505
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6406..6425
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
       61 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
      121 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
      181 attaatgcag aacggctccg cccactatta atgaaattaa aaattccaat tttaaaaaac
      241 gcagcaagag aaacatttgt atgaaagaat gcgtagaagg aaagaaaaat gtcgtcgaca
      301 tgctgaacaa caagattaat atgcctccgt gtataaaaaa aatattgaac gatttgaaag
      361 aaaacaatgt accgcgcggc ggtatgtaca ggaagaggtt tatactaaac tgttacattg
      421 caaacgtggt ttcgtgtgcc aagtgtgaaa accgatgttt aatcaaggct ctgacgcatt
      481 tctacaacca cgactccaag tgtgtgggtg aagtcatgca tcttttaatc aaatcccaag
      541 atgtgtataa accaccaaac tgccaaaaaa tgaaaactgt cgacaagctc tgtccgtttg
      601 ctggcaactg caagggtctc aatcctattt gtaattattg aataataaaa caattataaa
      661 tgtcaaattt gttttttatt aacgatacaa accaaacgca acaagaacat ttgtagtatt
      721 atctataatt gaaaacgcgt agttataatc gctgaggtaa tatttaaaat cattttcaaa
      781 tgattcacag ttaatttgcg acaatataat tttattttca cataaactag acgccttgtc
      841 gtcttcttct tcgtattcct tctctttttc atttttctcc tcataaaaat taacatagtt
      901 attatcgtat ccatatatgt atctatcgta tagagtaaat tttttgttgt cataaatata
      961 tatgtctttt ttaatggggt gtatagtacc gctgcgcata gtttttctgt aatttacaac
     1021 agtgctattt tctggtagtt cttcggagtg tgttgcttta attattaaat ttatataatc
     1081 aatgaatttg ggatcgtcgg ttttgtacaa tatgttgccg gcatagtacg cagcttcttc
     1141 tagttcaatt acaccatttt ttagcagcac cggattaaca taactttcca aaatgttgta
     1201 cgaaccgtta aacaaaaaca gttcacctcc cttttctata ctattgtctg cgagcagttg
     1261 tttgttgtta aaaataacag ccattgtaat gagacgcaca aactaatatc acaaactgga
     1321 aatgtctatc aatatatagt tgctgatatc ataacttcgt atagcataca ttatacgaag
     1381 ttatctggtt taaacgtacc cgtagtggct atggcagggc ttgccgcccc gacgttggct
     1441 gcgagccctg ggccttcacc cgaacttggg ggttggggtg gggaaaagga agaaacgcgg
     1501 gcgtattggt cccaatgggg tctcggtggg gtatcgacag agtgccagcc ctgggaccga
     1561 accccgcgtt tatgaacaaa cgacccaaca cccgtgcgtt ttattctgtc tttttattgc
     1621 cgtcatagcg cgggttcctt ccggtattgt ctccttccgt gtttcagtta gcctccccca
     1681 tctcccggta ccgcatgcta tgcatcagct gctagcacca tggctcgaga tcccgggtga
     1741 tcaagtcttc gtcgagtgat tgtaaataaa atgtaattta cagtatagta ttttaattaa
     1801 tatacaaatg atttgataat aattcttatt taactataat atattgtgtt gggttgaatt
     1861 aaaggtccgt atactagtat cgattcgcga cctactccgg aatattaata gatcatggag
     1921 ataattaaaa tgataaccat ctcgcaaata aataagtatt ttactgtttt cgtaacagtt
     1981 ttgtaataaa aaaacctata aatattccgg attattcata ccgtcccacc atcgggcgcg
     2041 gatcccggtc cgaagcgcgc ggaattcaaa ggcctacgtc gacgagctca ctagtcgcgg
     2101 ccgctttcga atctagagac tacaaggacg acgatgacaa gtaactgcag tctcgacaag
     2161 cttgtcgaga agtactagag gatcataatc agccatacca catttgtaga ggttttactt
     2221 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     2281 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2341 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2401 gtatcttatc atgtctggat ctgatcactg cttgagccta gaagatccgg ctgctaacaa
     2461 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactat cataacccct
     2521 aggcgattaa ttgatccggg ttattagtac atttattaag cgctagattc tgtgcgttgt
     2581 tgatttacag acaattgttg tacgtatttt aataattcat taaatttata atctttaggg
     2641 tggtatgtta gagcgaaaat caaatgattt tcagcgtctt tatatctgaa tttaaatatt
     2701 aaatcctcaa tagatttgta aaataggttt cgattagttt caaacaaggg ttgtttttcc
     2761 gaaccgatgg ctggactatc taatggattt tcgctcaacg ccacaaaact tgccaaatct
     2821 tgtagcagca atctagcttt gtcgatattc gtttgtgttt tgttttgtaa taaaggttcg
     2881 acgtcgttca aaatattatg cgcttttgta tttctttcat cactgtcgtt agtgtacaat
     2941 tgactcgacg taaacacgtt aaataaagct tggacatatt taacatcggg cgtgttagct
     3001 ttattaggcc gattatcgtc gtcgtcccaa ccctcgtcgt tagaagttgc ttccgaagac
     3061 gattttgcca tagccacacg acgcctatta attgtgtcgg ctaacacgtc cgcgatcaaa
     3121 tttgtagttg agctttttgg aattatttct gattgcgggc gtttttgggc gggtttcaat
     3181 ctaactgtgc ccgattttaa ttcagacaac acgttagaaa gcgatggtgc aggcggtggt
     3241 aacatttcag acggcaaatc tactaatggc ggcggtggtg gagctgatga taaatctacc
     3301 atcggtggag gcgcaggcgg ggctggcggc ggaggcggag gcggaggtgg tggcggtgat
     3361 gcagacggcg gtttaggctc aaatgtctct ttaggcaaca cagtcggcac ctcaactatt
     3421 gtactggttt cgggcgccgt ttttggtttg accggtctga gacgagtgcg atttttttcg
     3481 tttctaatag cttccaacaa ttgttgtctg tcgtctaaag gtgcagcggg ttgaggttcc
     3541 gtcggcattg gtggagcggg cggcaattca gacatcgatg gtggtggtgg tggtggaggc
     3601 gctggaatgt taggcacggg agaaggtggt ggcggcggtg ccgccggtat aatttgttct
     3661 ggtttagttt gttcgcgcac gattgtgggc accggcgcag gcgccgctgg ctgcacaacg
     3721 gaaggtcgtc tgcttcgagg cagcgcttgg ggtggtggca attcaatatt ataattggaa
     3781 tacaaatcgt aaaaatctgc tataagcatt gtaatttcgc tatcgtttac cgtgccgata
     3841 tttaacaacc gctcaatgta agcaattgta ttgtaaagag attgtctcaa gctcggatcg
     3901 atcccgcacg ccgataacaa gccttttcat ttttactaca gcattgtagt ggcgagacac
     3961 ttcgctgtcg tcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4021 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4081 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4141 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4201 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     4261 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4321 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4381 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4441 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     4501 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4561 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4621 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4681 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4741 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4801 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4861 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4921 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4981 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5041 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5101 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5161 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5221 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5281 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5341 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5401 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5461 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5521 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5581 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5641 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5701 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5761 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5821 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5881 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5941 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6001 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6061 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6121 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6181 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6241 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6301 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6361 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6421 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6481 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6541 cttttgctgg c
//
