LOCUS       sequence-379027-        7362 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 6 His tag under the pH promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7362)
  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 7362)
  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-6His-Nter-GWs-Lox (VE5587)
FEATURES             Location/Qualifiers
     source          1..7362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      689..1144
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(776..795)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     986..1007
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1161..1180)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1280..1302
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1340..1358)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1426..1530
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1531..1599,1600..2391)
                     /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(1749..1768)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2562..3150
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3051..3070
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3304..3321
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_recomb     3713..4540
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             3713..3837
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(3875..4480)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     protein_bind    complement(4544..4577)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4578..4669
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4594..4611
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4693..4710
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4720..4743
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     protein_bind    4768..4892
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(4917..4922,4923..4940,4941..4947)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             5001..5660
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5067..5086)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             6002..6307
                     /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     6186..6205
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(6348..6472)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             6480..6509
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_recomb     join(6547..7362,1..542)
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(join(6563..7362,1..542))
                     /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 gtgatgcaga cggcggttta ggctcaaatg tctctttagg caacacagtc ggcacctcaa
       61 ctattgtact ggtttcgggc gccgtttttg gtttgaccgg tctgagacga gtgcgatttt
      121 tttcgtttct aatagcttcc aacaattgtt gtctgtcgtc taaaggtgca gcgggttgag
      181 gttccgtcgg cattggtgga gcgggcggca attcagacat cgatggtggt ggtggtggtg
      241 gaggcgctgg aatgttaggc acgggagaag gtggtggcgg cggtgccgcc ggtataattt
      301 gttctggttt agtttgttcg cgcacgattg tgggcaccgg cgcaggcgcc gctggctgca
      361 caacggaagg tcgtctgctt cgaggcagcg cttggggtgg tggcaattca atattataat
      421 tggaatacaa atcgtaaaaa tctgctataa gcattgtaat ttcgctatcg tttaccgtgc
      481 cgatatttaa caaccgctca atgtaagcaa ttgtattgta aagagattgt ctcaagctcg
      541 gatcgatccc gcacgccgat aacaagcctt ttcattttta ctacagcatt gtagtggcga
      601 gacacttcgc tgtcgtcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
      661 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
      721 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
      781 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
      841 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
      901 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
      961 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     1021 cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     1081 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     1141 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     1201 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     1261 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     1321 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     1381 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1441 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1501 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     1561 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1621 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1681 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1741 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     1801 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1861 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     1921 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     1981 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     2041 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     2101 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2161 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2221 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     2281 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     2341 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     2401 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2461 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2521 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2581 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2641 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2701 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2761 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2821 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2881 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2941 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3001 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3061 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3121 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3181 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3241 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3301 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     3361 gcgcgttggc cgattcatta atgcagaacg gctccgccca ctattaatga aattaaaaat
     3421 tccaatttta aaaaacgcag caagagaaac atttgtatga aagaatgcgt agaaggaaag
     3481 aaaaatgtcg tcgacatgct gaacaacaag attaatatgc ctccgtgtat aaaaaaaata
     3541 ttgaacgatt tgaaagaaaa caatgtaccg cgcggcggta tgtacaggaa gaggtttata
     3601 ctaaactgtt acattgcaaa cgtggtttcg tgtgccaagt gtgaaaaccg atgtttaatc
     3661 aaggctctga cgcatttcta caaccacgac tccaagtgtg tgggtgaagt catgcatctt
     3721 ttaatcaaat cccaagatgt gtataaacca ccaaactgcc aaaaaatgaa aactgtcgac
     3781 aagctctgtc cgtttgctgg caactgcaag ggtctcaatc ctatttgtaa ttattgaata
     3841 ataaaacaat tataaatgtc aaatttgttt tttattaacg atacaaacca aacgcaacaa
     3901 gaacatttgt agtattatct ataattgaaa acgcgtagtt ataatcgctg aggtaatatt
     3961 taaaatcatt ttcaaatgat tcacagttaa tttgcgacaa tataatttta ttttcacata
     4021 aactagacgc cttgtcgtct tcttcttcgt attccttctc tttttcattt ttctcctcat
     4081 aaaaattaac atagttatta tcgtatccat atatgtatct atcgtataga gtaaattttt
     4141 tgttgtcata aatatatatg tcttttttaa tggggtgtat agtaccgctg cgcatagttt
     4201 ttctgtaatt tacaacagtg ctattttctg gtagttcttc ggagtgtgtt gctttaatta
     4261 ttaaatttat ataatcaatg aatttgggat cgtcggtttt gtacaatatg ttgccggcat
     4321 agtacgcagc ttcttctagt tcaattacac cattttttag cagcaccgga ttaacataac
     4381 tttccaaaat gttgtacgaa ccgttaaaca aaaacagttc acctcccttt tctatactat
     4441 tgtctgcgag cagttgtttg ttgttaaaaa taacagccat tgtaatgaga cgcacaaact
     4501 aatatcacaa actggaaatg tctatcaata tatagttgct gatataactt cgtatagcat
     4561 acattatacg aagttatatc atggagataa ttaaaatgat aaccatctcg caaataaata
     4621 agtattttac tgttttcgta acagttttgt aataaaaaaa cctataaata cggatctacc
     4681 atgggcagca gccatcatca tcatcatcac tccgcgggtc tggaagttct gttccagggg
     4741 ccccatatgg gatccgaatt cccatcaaca agtttgtaca aaaaagctga acgagaaacg
     4801 taaaatgata taaatatcaa tatattaaat tagattttgc ataaaaaaca gactacataa
     4861 tactgtaaaa cacaacatat ccagtcacta tggcggccgc attaggcacc ccaggcttta
     4921 cactttatgc ttccggctcg tataatgtgt ggattttgag ttaggatccg tcgagatttt
     4981 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat
     5041 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata
     5101 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca
     5161 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc
     5221 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg
     5281 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc
     5341 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt
     5401 tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca
     5461 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg
     5521 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg
     5581 ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg
     5641 agtggcaggg cggggcgtaa agatctggat ccggcttact aaaagccaga taacagtatg
     5701 cgtatttgcg cgctgatttt tgcggtataa gaatatatac tgatatgtat acccgaagta
     5761 tgtcaaaaag aggtatgcta tgaagcagcg tattacagtg acagttgaca gcgacagcta
     5821 tcagttgctc aaggcatata tgatgtcaat atctccggtc tggtaagcac aaccatgcag
     5881 aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga agggatggct
     5941 gaggtcgccc ggtttattga aatgaacggc tcttttgctg acgagaacag gggctggtga
     6001 aatgcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt
     6061 acagagtgat attattgaca cgcccgggcg acggatggtg atccccctgg ccagtgcacg
     6121 tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag
     6181 ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt
     6241 ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg
     6301 aatataaatg tcaggctccc ttatacacag ccagtctgca ggtcgaccat agtgactgga
     6361 tatgttgtgt tttacagtat tatgtagtct gttttttatg caaaatctaa tttaatatat
     6421 tgatatttat atcattttac gtttctcgtt cagctttctt gtacaaagtg gtaggatccg
     6481 aacaaaaact catctcagaa gaggatctgt aagcttgatg ggcggccgct taattaattg
     6541 atccgggtta ttagtacatt tattaagcgc tagattctgt gcgttgttga tttacagaca
     6601 attgttgtac gtattttaat aattcattaa atttataatc tttagggtgg tatgttagag
     6661 cgaaaatcaa atgattttca gcgtctttat atctgaattt aaatattaaa tcctcaatag
     6721 atttgtaaaa taggtttcga ttagtttcaa acaagggttg tttttccgaa ccgatggctg
     6781 gactatctaa tggattttcg ctcaacgcca caaaacttgc caaatcttgt agcagcaatc
     6841 tagctttgtc gatattcgtt tgtgttttgt tttgtaataa aggttcgacg tcgttcaaaa
     6901 tattatgcgc ttttgtattt ctttcatcac tgtcgttagt gtacaattga ctcgacgtaa
     6961 acacgttaaa taaagcttgg acatatttaa catcgggcgt gttagcttta ttaggccgat
     7021 tatcgtcgtc gtcccaaccc tcgtcgttag aagttgcttc cgaagacgat tttgccatag
     7081 ccacacgacg cctattaatt gtgtcggcta acacgtccgc gatcaaattt gtagttgagc
     7141 tttttggaat tatttctgat tgcgggcgtt tttgggcggg tttcaatcta actgtgcccg
     7201 attttaattc agacaacacg ttagaaagcg atggtgcagg cggtggtaac atttcagacg
     7261 gcaaatctac taatggcggc ggtggtggag ctgatgataa atctaccatc ggtggaggcg
     7321 caggcggggc tggcggcgga ggcggaggcg gaggtggtgg cg
//
