LOCUS       sequence-270511-        6937 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 10-His tag under the pH promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6937)
  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 6937)
  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-3C-10His-Cter (VE5622)
FEATURES             Location/Qualifiers
     source          1..6937
                     /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"
     promoter        complement(408..517)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        562..653
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     578..595
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             768..791
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             792..821
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     polyA_signal    967..1101
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1004..1023)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1058..1077
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_recomb     1220..2577
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(1236..2577)
                     /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      2724..3179
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2811..2830)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3021..3042
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3196..3215)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3315..3337
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3375..3393)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3461..3565
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3566..3634,3635..4426)
                     /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(3784..3803)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4597..5185
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5086..5105
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5339..5356
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_recomb     5748..6575
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             5748..5872
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5910..6515)
                     /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 ccgcatgcta tgcatcagct
      361 gctagcacca tggctcgaga tcccgggtga tcaagtcttc gtcgagtgat tgtaaataaa
      421 atgtaattta cagtatagta ttttaattaa tatacaaatg atttgataat aattcttatt
      481 taactataat atattgtgtt gggttgaatt aaaggtccgt atactagtat cgattcgcga
      541 cctactccgg aatattaata gatcatggag ataattaaaa tgataaccat ctcgcaaata
      601 aataagtatt ttactgtttt cgtaacagtt ttgtaataaa aaaacctata aatattccgg
      661 attattcata ccgtcccacc atcgggcgcg gatcccggtc cgaagcgcgc ggaattcaaa
      721 ggcctacgtc gacgagctca ctagtcgcgg ccgctttcga atctagactg gaagttctgt
      781 ttcagggtcc gcaccatcat caccatcacc atcaccacca ttaactgcag tctcgacaag
      841 cttgtcgaga agtactagag gatcataatc agccatacca catttgtaga ggttttactt
      901 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
      961 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     1021 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     1081 gtatcttatc atgtctggat ctgatcactg cttgagccta gaagatccgg ctgctaacaa
     1141 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactat cataacccct
     1201 aggcgattaa ttgatccggg ttattagtac atttattaag cgctagattc tgtgcgttgt
     1261 tgatttacag acaattgttg tacgtatttt aataattcat taaatttata atctttaggg
     1321 tggtatgtta gagcgaaaat caaatgattt tcagcgtctt tatatctgaa tttaaatatt
     1381 aaatcctcaa tagatttgta aaataggttt cgattagttt caaacaaggg ttgtttttcc
     1441 gaaccgatgg ctggactatc taatggattt tcgctcaacg ccacaaaact tgccaaatct
     1501 tgtagcagca atctagcttt gtcgatattc gtttgtgttt tgttttgtaa taaaggttcg
     1561 acgtcgttca aaatattatg cgcttttgta tttctttcat cactgtcgtt agtgtacaat
     1621 tgactcgacg taaacacgtt aaataaagct tggacatatt taacatcggg cgtgttagct
     1681 ttattaggcc gattatcgtc gtcgtcccaa ccctcgtcgt tagaagttgc ttccgaagac
     1741 gattttgcca tagccacacg acgcctatta attgtgtcgg ctaacacgtc cgcgatcaaa
     1801 tttgtagttg agctttttgg aattatttct gattgcgggc gtttttgggc gggtttcaat
     1861 ctaactgtgc ccgattttaa ttcagacaac acgttagaaa gcgatggtgc aggcggtggt
     1921 aacatttcag acggcaaatc tactaatggc ggcggtggtg gagctgatga taaatctacc
     1981 atcggtggag gcgcaggcgg ggctggcggc ggaggcggag gcggaggtgg tggcggtgat
     2041 gcagacggcg gtttaggctc aaatgtctct ttaggcaaca cagtcggcac ctcaactatt
     2101 gtactggttt cgggcgccgt ttttggtttg accggtctga gacgagtgcg atttttttcg
     2161 tttctaatag cttccaacaa ttgttgtctg tcgtctaaag gtgcagcggg ttgaggttcc
     2221 gtcggcattg gtggagcggg cggcaattca gacatcgatg gtggtggtgg tggtggaggc
     2281 gctggaatgt taggcacggg agaaggtggt ggcggcggtg ccgccggtat aatttgttct
     2341 ggtttagttt gttcgcgcac gattgtgggc accggcgcag gcgccgctgg ctgcacaacg
     2401 gaaggtcgtc tgcttcgagg cagcgcttgg ggtggtggca attcaatatt ataattggaa
     2461 tacaaatcgt aaaaatctgc tataagcatt gtaatttcgc tatcgtttac cgtgccgata
     2521 tttaacaacc gctcaatgta agcaattgta ttgtaaagag attgtctcaa gctcggatcg
     2581 atcccgcacg ccgataacaa gccttttcat ttttactaca gcattgtagt ggcgagacac
     2641 ttcgctgtcg tcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2701 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2761 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2821 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2881 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     2941 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3001 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3061 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3121 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     3181 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3241 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3301 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3361 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3421 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3481 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3541 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3601 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3661 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3721 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3781 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3841 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3901 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3961 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4021 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4081 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4141 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4201 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4261 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4321 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4381 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4441 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4501 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4561 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4621 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4681 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4741 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4801 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4861 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4921 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4981 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5041 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5101 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5161 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5221 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     5281 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     5341 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     5401 ttggccgatt cattaatgca gaacggctcc gcccactatt aatgaaatta aaaattccaa
     5461 ttttaaaaaa cgcagcaaga gaaacatttg tatgaaagaa tgcgtagaag gaaagaaaaa
     5521 tgtcgtcgac atgctgaaca acaagattaa tatgcctccg tgtataaaaa aaatattgaa
     5581 cgatttgaaa gaaaacaatg taccgcgcgg cggtatgtac aggaagaggt ttatactaaa
     5641 ctgttacatt gcaaacgtgg tttcgtgtgc caagtgtgaa aaccgatgtt taatcaaggc
     5701 tctgacgcat ttctacaacc acgactccaa gtgtgtgggt gaagtcatgc atcttttaat
     5761 caaatcccaa gatgtgtata aaccaccaaa ctgccaaaaa atgaaaactg tcgacaagct
     5821 ctgtccgttt gctggcaact gcaagggtct caatcctatt tgtaattatt gaataataaa
     5881 acaattataa atgctaaatt tgttttttat taacgataca aaccaaacgc aacaagaaca
     5941 tttgtagtat tatctataat tgaaaacgcg tagttataat cgctgaggta atatttaaaa
     6001 tcattttcaa atgattcaca gttaatttgc gacaatataa ttttattttc acataaacta
     6061 gacgccttgt cgtcttcttc ttcgtattcc ttctcttttt catttttctc ctcataaaaa
     6121 ttaacatagt tattatcgta tccatatatg tatctatcgt atagagtaaa ttttttgttg
     6181 tcataaatat atatgtcttt tttaatgggg tgtatagtac cgctgcgcat agtttttctg
     6241 taatttacaa cagtgctatt ttctggtagt tcttcggagt gtgttgcttt aattattaaa
     6301 tttatataat caatgaattt gggatcgtcg gttttgtaca atatgttgcc ggcatagtac
     6361 gcagcttctt ctagttcaat tacaccattt tttagcagca ccggattaac ataactttcc
     6421 aaaatgttgt acgaaccgtt aaacaaaaac agttcacctc ccttttctat actattgtct
     6481 gcgagcagtt gtttgttgtt aaaaataaca gccattgtaa tgagacgcac aaactaatat
     6541 cacaaactgg aaatgtctat caatatatag ttgctgatat cccattttac caccataata
     6601 cttaaaatga tccacaaaca agccaccacc aatcgctaca ctcaccgccg ctacagacgc
     6661 tacaatcacc tcatatttat tctcctcaca tatttattct attctaattc tcaatcatac
     6721 caatcaactt ctcatactca atcataccaa tcaatacaca ttatactact actcatttcc
     6781 aatcattcac attcataata tatctgttat actactcctt atcactactc tctccatcca
     6841 cattattatc ctccatcata cgacgctaca taactacgtg gttcctaact acaccaagca
     6901 tagccctagc cagccagcca caccttacag ccttttt
//
