LOCUS       sequence-324175-        6535 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 p10 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6535)
  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 6535)
  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-Flag-Cter(VE5738)
FEATURES             Location/Qualifiers
     source          1..6535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     52..74
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(112..130)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        198..302
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(303..371,372..1163)
                     /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(521..540)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1334..1922
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1823..1842
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2076..2093
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_recomb     2485..3312
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             2485..2609
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(2647..3252)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     protein_bind    complement(3319..3352)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3542..3590
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3615..3634
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3664..3687)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3735..3844)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        3889..3980
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3905..3922
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    4239..4373
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4276..4295)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4330..4349
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_recomb     4492..5849
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(4508..5849)
                     /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      5996..6451
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6083..6102)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6293..6314
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6468..6487)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
       61 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
      121 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
      181 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
      241 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
      301 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
      361 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
      421 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
      481 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
      541 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
      601 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
      661 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
      721 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
      781 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
      841 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
      901 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
      961 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     1021 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     1081 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     1141 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     1201 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     1261 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     1321 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1381 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1441 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1501 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1561 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1621 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1681 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1741 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1801 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1861 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     1921 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     1981 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     2041 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     2101 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagaa
     2161 cggctccgcc cactattaat gaaattaaaa attccaattt taaaaaacgc agcaagagaa
     2221 acatttgtat gaaagaatgc gtagaaggaa agaaaaatgt cgtcgacatg ctgaacaaca
     2281 agattaatat gcctccgtgt ataaaaaaaa tattgaacga tttgaaagaa aacaatgtac
     2341 cgcgcggcgg tatgtacagg aagaggttta tactaaactg ttacattgca aacgtggttt
     2401 cgtgtgccaa gtgtgaaaac cgatgtttaa tcaaggctct gacgcatttc tacaaccacg
     2461 actccaagtg tgtgggtgaa gtcatgcatc ttttaatcaa atcccaagat gtgtataaac
     2521 caccaaactg ccaaaaaatg aaaactgtcg acaagctctg tccgtttgct ggcaactgca
     2581 agggtctcaa tcctatttgt aattattgaa taataaaaca attataaatg tcaaatttgt
     2641 tttttattaa cgatacaaac caaacgcaac aagaacattt gtagtattat ctataattga
     2701 aaacgcgtag ttataatcgc tgaggtaata tttaaaatca ttttcaaatg attcacagtt
     2761 aatttgcgac aatataattt tattttcaca taaactagac gccttgtcgt cttcttcttc
     2821 gtattccttc tctttttcat ttttctcctc ataaaaatta acatagttat tatcgtatcc
     2881 atatatgtat ctatcgtata gagtaaattt tttgttgtca taaatatata tgtctttttt
     2941 aatggggtgt atagtaccgc tgcgcatagt ttttctgtaa tttacaacag tgctattttc
     3001 tggtagttct tcggagtgtg ttgctttaat tattaaattt atataatcaa tgaatttggg
     3061 atcgtcggtt ttgtacaata tgttgccggc atagtacgca gcttcttcta gttcaattac
     3121 accatttttt agcagcaccg gattaacata actttccaaa atgttgtacg aaccgttaaa
     3181 caaaaacagt tcacctccct tttctatact attgtctgcg agcagttgtt tgttgttaaa
     3241 aataacagcc attgtaatga gacgcacaaa ctaatatcac aaactggaaa tgtctatcaa
     3301 tatatagttg ctgatatcat aacttcgtat agcatacatt atacgaagtt atctggttta
     3361 aacgtacccg tagtggctat ggcagggctt gccgccccga cgttggctgc gagccctggg
     3421 ccttcacccg aacttggggg ttggggtggg gaaaaggaag aaacgcgggc gtattggtcc
     3481 caatggggtc tcggtggggt atcgacagag tgccagccct gggaccgaac cccgcgttta
     3541 tgaacaaacg acccaacacc cgtgcgtttt attctgtctt tttattgccg tcatagcgcg
     3601 ggttccttcc ggtattgtct ccttccgtgt ttcagttagc ctcccccatc tcccggtacc
     3661 ttacttgtca tcgtcgtcct tgtagtcgct agcaccatgg ctcgagatcc cgggtgatca
     3721 agtcttcgtc gagtgattgt aaataaaatg taatttacag tatagtattt taattaatat
     3781 acaaatgatt tgataataat tcttatttaa ctataatata ttgtgttggg ttgaattaaa
     3841 ggtccgtata ctagtatcga ttcgcgacct actccggaat attaatagat catggagata
     3901 attaaaatga taaccatctc gcaaataaat aagtatttta ctgttttcgt aacagttttg
     3961 taataaaaaa acctataaat attccggatt attcataccg tcccaccatc gggcgcggat
     4021 cccggtccga agcgcgcgga attcaaaggc ctacgtcgac gagctcacta gtcgcggccg
     4081 ctttcgaatc tagagcctgc agtctcgaca agcttgtcga gaagtactag aggatcataa
     4141 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     4201 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     4261 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4321 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctgatcac
     4381 tgcttgagcc tagaagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     4441 gccaccgctg agcaataact atcataaccc ctaggcgatt aattgatccg ggttattagt
     4501 acatttatta agcgctagat tctgtgcgtt gttgatttac agacaattgt tgtacgtatt
     4561 ttaataattc attaaattta taatctttag ggtggtatgt tagagcgaaa atcaaatgat
     4621 tttcagcgtc tttatatctg aatttaaata ttaaatcctc aatagatttg taaaataggt
     4681 ttcgattagt ttcaaacaag ggttgttttt ccgaaccgat ggctggacta tctaatggat
     4741 tttcgctcaa cgccacaaaa cttgccaaat cttgtagcag caatctagct ttgtcgatat
     4801 tcgtttgtgt tttgttttgt aataaaggtt cgacgtcgtt caaaatatta tgcgcttttg
     4861 tatttctttc atcactgtcg ttagtgtaca attgactcga cgtaaacacg ttaaataaag
     4921 cttggacata tttaacatcg ggcgtgttag ctttattagg ccgattatcg tcgtcgtccc
     4981 aaccctcgtc gttagaagtt gcttccgaag acgattttgc catagccaca cgacgcctat
     5041 taattgtgtc ggctaacacg tccgcgatca aatttgtagt tgagcttttt ggaattattt
     5101 ctgattgcgg gcgtttttgg gcgggtttca atctaactgt gcccgatttt aattcagaca
     5161 acacgttaga aagcgatggt gcaggcggtg gtaacatttc agacggcaaa tctactaatg
     5221 gcggcggtgg tggagctgat gataaatcta ccatcggtgg aggcgcaggc ggggctggcg
     5281 gcggaggcgg aggcggaggt ggtggcggtg atgcagacgg cggtttaggc tcaaatgtct
     5341 ctttaggcaa cacagtcggc acctcaacta ttgtactggt ttcgggcgcc gtttttggtt
     5401 tgaccggtct gagacgagtg cgattttttt cgtttctaat agcttccaac aattgttgtc
     5461 tgtcgtctaa aggtgcagcg ggttgaggtt ccgtcggcat tggtggagcg ggcggcaatt
     5521 cagacatcga tggtggtggt ggtggtggag gcgctggaat gttaggcacg ggagaaggtg
     5581 gtggcggcgg tgccgccggt ataatttgtt ctggtttagt ttgttcgcgc acgattgtgg
     5641 gcaccggcgc aggcgccgct ggctgcacaa cggaaggtcg tctgcttcga ggcagcgctt
     5701 ggggtggtgg caattcaata ttataattgg aatacaaatc gtaaaaatct gctataagca
     5761 ttgtaatttc gctatcgttt accgtgccga tatttaacaa ccgctcaatg taagcaattg
     5821 tattgtaaag agattgtctc aagctcggat cgatcccgca cgccgataac aagccttttc
     5881 atttttacta cagcattgta gtggcgagac acttcgctgt cgtcgcctga tgcggtattt
     5941 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     6001 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     6061 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     6121 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     6181 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     6241 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6301 ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg
     6361 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6421 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     6481 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccct
//
