LOCUS       sequence-324125-        7374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transfer vector for gene expression to generate recombinant 
            baculoviruses by homologous recombination. Contains expression 
            cassette with an N-ter 10 His tag under the pH promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7374)
  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 7374)
  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_PH-10His-Nter-GWs-Lox (VE5588)
FEATURES             Location/Qualifiers
     source          1..7374
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1440..1895
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1527..1546)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1737..1758
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1912..1931)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2031..2053
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2091..2109)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2177..2281
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2282..2350,2351..3142)
                     /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(2500..2519)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3313..3901
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3802..3821
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4055..4072
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_recomb     4464..5291
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             4464..4588
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4626..5231)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     protein_bind    complement(5295..5328)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5329..5420
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5345..5362
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             5444..5473
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             5483..5506
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     protein_bind    5531..5655
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(5680..5685,5686..5703,5704..5710)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             5764..6423
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5830..5849)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             6765..7070
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     6949..6968
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(7111..7235)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             7243..7272
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_recomb     join(7310..7374,1..1293)
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(join(7326..7374,1..1293))
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="PSLRQSLYNTIAYIERLLNIGTVNDSEITMLIADFYDLYSNYNIE
                     LPPPQALPRSRRPSVVQPAAPAPVPTIVREQTKPEQIIPAAPPPPPSPVPNIPAPPPPP
                     PPSMSELPPAPPMPTEPQPAAPLDDRQQLLEAIRNEKNRTRLRPVKPKTAPETSTIVEV
                     PTVLPKETFEPKPPSASPPPPPPPPPPPAPPAPPPMVDLSSAPPPPPLVDLPSEMLPPP
                     APSLSNVLSELKSGTVRLKPAQKRPQSEIIPKSSTTNLIADVLADTINRRRVAMAKSSS
                     EATSNDEGWDDDDNRPNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVE
                     PLLQNKTQTNIDKARLLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDL
                     IFKFRYKDAENHLIFALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
ORIGIN
        1 tattttaata attcattaaa tttataatct ttagggtggt atgttagagc gaaaatcaaa
       61 tgattttcag cgtctttata tctgaattta aatattaaat cctcaataga tttgtaaaat
      121 aggtttcgat tagtttcaaa caagggttgt ttttccgaac cgatggctgg actatctaat
      181 ggattttcgc tcaacgccac aaaacttgcc aaatcttgta gcagcaatct agctttgtcg
      241 atattcgttt gtgttttgtt ttgtaataaa ggttcgacgt cgttcaaaat attatgcgct
      301 tttgtatttc tttcatcact gtcgttagtg tacaattgac tcgacgtaaa cacgttaaat
      361 aaagcttgga catatttaac atcgggcgtg ttagctttat taggccgatt atcgtcgtcg
      421 tcccaaccct cgtcgttaga agttgcttcc gaagacgatt ttgccatagc cacacgacgc
      481 ctattaattg tgtcggctaa cacgtccgcg atcaaatttg tagttgagct ttttggaatt
      541 atttctgatt gcgggcgttt ttgggcgggt ttcaatctaa ctgtgcccga ttttaattca
      601 gacaacacgt tagaaagcga tggtgcaggc ggtggtaaca tttcagacgg caaatctact
      661 aatggcggcg gtggtggagc tgatgataaa tctaccatcg gtggaggcgc aggcggggct
      721 ggcggcggag gcggaggcgg aggtggtggc ggtgatgcag acggcggttt aggctcaaat
      781 gtctctttag gcaacacagt cggcacctca actattgtac tggtttcggg cgccgttttt
      841 ggtttgaccg gtctgagacg agtgcgattt ttttcgtttc taatagcttc caacaattgt
      901 tgtctgtcgt ctaaaggtgc agcgggttga ggttccgtcg gcattggtgg agcgggcggc
      961 aattcagaca tcgatggtgg tggtggtggt ggaggcgctg gaatgttagg cacgggagaa
     1021 ggtggtggcg gcggtgccgc cggtataatt tgttctggtt tagtttgttc gcgcacgatt
     1081 gtgggcaccg gcgcaggcgc cgctggctgc acaacggaag gtcgtctgct tcgaggcagc
     1141 gcttggggtg gtggcaattc aatattataa ttggaataca aatcgtaaaa atctgctata
     1201 agcattgtaa tttcgctatc gtttaccgtg ccgatattta acaaccgctc aatgtaagca
     1261 attgtattgt aaagagattg tctcaagctc ggatcgatcc cgcacgccga taacaagcct
     1321 tttcattttt actacagcat tgtagtggcg agacacttcg ctgtcgtcgc ctgatgcggt
     1381 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     1441 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1501 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     1561 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1621 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     1681 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1741 actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata
     1801 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1861 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     1921 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     1981 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     2041 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     2101 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     2161 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2221 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2281 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2341 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     2401 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     2461 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     2521 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     2581 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2641 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2701 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2761 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     2821 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     2881 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     2941 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3001 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3061 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3121 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3181 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3241 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3301 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3361 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3421 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3481 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3541 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3601 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3661 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3721 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3781 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3841 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3901 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3961 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     4021 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     4081 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagaac
     4141 ggctccgccc actattaatg aaattaaaaa ttccaatttt aaaaaacgca gcaagagaaa
     4201 catttgtatg aaagaatgcg tagaaggaaa gaaaaatgtc gtcgacatgc tgaacaacaa
     4261 gattaatatg cctccgtgta taaaaaaaat attgaacgat ttgaaagaaa acaatgtacc
     4321 gcgcggcggt atgtacagga agaggtttat actaaactgt tacattgcaa acgtggtttc
     4381 gtgtgccaag tgtgaaaacc gatgtttaat caaggctctg acgcatttct acaaccacga
     4441 ctccaagtgt gtgggtgaag tcatgcatct tttaatcaaa tcccaagatg tgtataaacc
     4501 accaaactgc caaaaaatga aaactgtcga caagctctgt ccgtttgctg gcaactgcaa
     4561 gggtctcaat cctatttgta attattgaat aataaaacaa ttataaatgt caaatttgtt
     4621 ttttattaac gatacaaacc aaacgcaaca agaacatttg tagtattatc tataattgaa
     4681 aacgcgtagt tataatcgct gaggtaatat ttaaaatcat tttcaaatga ttcacagtta
     4741 atttgcgaca atataatttt attttcacat aaactagacg ccttgtcgtc ttcttcttcg
     4801 tattccttct ctttttcatt tttctcctca taaaaattaa catagttatt atcgtatcca
     4861 tatatgtatc tatcgtatag agtaaatttt ttgttgtcat aaatatatat gtctttttta
     4921 atggggtgta tagtaccgct gcgcatagtt tttctgtaat ttacaacagt gctattttct
     4981 ggtagttctt cggagtgtgt tgctttaatt attaaattta tataatcaat gaatttggga
     5041 tcgtcggttt tgtacaatat gttgccggca tagtacgcag cttcttctag ttcaattaca
     5101 ccatttttta gcagcaccgg attaacataa ctttccaaaa tgttgtacga accgttaaac
     5161 aaaaacagtt cacctccctt ttctatacta ttgtctgcga gcagttgttt gttgttaaaa
     5221 ataacagcca ttgtaatgag acgcacaaac taatatcaca aactggaaat gtctatcaat
     5281 atatagttgc tgatataact tcgtatagca tacattatac gaagttatat catggagata
     5341 attaaaatga taaccatctc gcaaataaat aagtatttta ctgttttcgt aacagttttg
     5401 taataaaaaa acctataaat acggatctac catgggcagc agccaccatc accatcatca
     5461 tcatcatcat cactccgcgg gtctggaagt tctgttccag gggccccata tgggatccga
     5521 attcccatca acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg atataaatat
     5581 caatatatta aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca
     5641 tatccagtca ctatggcggc cgcattaggc accccaggct ttacacttta tgcttccggc
     5701 tcgtataatg tgtggatttt gagttaggat ccgtcgagat tttcaggagc taaggaagct
     5761 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa
     5821 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg
     5881 gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt
     5941 attcacattc ttgcccgcct gatgaatgct catccggaat tccgtatggc aatgaaagac
     6001 ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact
     6061 gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata
     6121 tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt
     6181 gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac
     6241 gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa
     6301 ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc
     6361 catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg
     6421 taaagatctg gatccggctt actaaaagcc agataacagt atgcgtattt gcgcgctgat
     6481 ttttgcggta taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtatg
     6541 ctatgaagca gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat
     6601 atatgatgtc aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct
     6661 gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat
     6721 tgaaatgaac ggctcttttg ctgacgagaa caggggctgg tgaaatgcag tttaaggttt
     6781 acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg
     6841 acacgcccgg gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag
     6901 tctcccgtga actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca
     6961 ccgatatggc cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc
     7021 gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct
     7081 cccttataca cagccagtct gcaggtcgac catagtgact ggatatgttg tgttttacag
     7141 tattatgtag tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt
     7201 tacgtttctc gttcagcttt cttgtacaaa gtggtaggat ccgaacaaaa actcatctca
     7261 gaagaggatc tgtaagcttg atgggcggcc gcttaattaa ttgatccggg ttattagtac
     7321 atttattaag cgctagattc tgtgcgttgt tgatttacag acaattgttg tacg
//
