LOCUS       sequence-379026-        7283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transfer vector for gene expression to generate recombinant 
            baculoviruses by homologous recombination. Contains expression 
            cassette under the pH promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7283)
  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 7283)
  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_GWs-Lox (VE5589)
FEATURES             Location/Qualifiers
     source          1..7283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      354..809
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(441..460)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     651..672
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(826..845)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     945..967
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1005..1023)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1091..1195
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1196..1264,1265..2056)
                     /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(1414..1433)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2227..2815
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2716..2735
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2969..2986
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_recomb     3378..4205
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             3378..3502
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(3540..4145)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     protein_bind    4209..4242
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4243..4334
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4259..4276
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     protein_bind    4355..4479
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(4504..4509,4510..4527,4528..4534)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             4588..5268
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(4654..4673)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             5588..5893
                     /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
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     5772..5791
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(5934..6058)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             6066..6095
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_recomb     join(6133..7283,1..207)
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(join(6149..7283,1..207))
                     /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 accggcgcag gcgccgctgg ctgcacaacg gaaggtcgtc tgcttcgagg cagcgcttgg
       61 ggtggtggca attcaatatt ataattggaa tacaaatcgt aaaaatctgc tataagcatt
      121 gtaatttcgc tatcgtttac cgtgccgata tttaacaacc gctcaatgta agcaattgta
      181 ttgtaaagag attgtctcaa gctcggatcg atcccgcacg ccgataacaa gccttttcat
      241 ttttactaca gcattgtagt ggcgagacac ttcgctgtcg tcgcctgatg cggtattttc
      301 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
      361 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
      421 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
      481 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
      541 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
      601 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
      661 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
      721 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
      781 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
      841 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
      901 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
      961 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     1021 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     1081 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1141 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1201 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1261 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1321 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1381 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     1441 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     1501 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     1561 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1621 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1681 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     1741 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     1801 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     1861 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     1921 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     1981 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2041 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2101 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2161 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2221 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2281 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2341 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     2401 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2461 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2521 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2581 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2641 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2701 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2761 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2821 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     2881 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     2941 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3001 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gaacggctcc
     3061 gcccactatt aatgaaatta aaaattccaa ttttaaaaaa cgcagcaaga gaaacatttg
     3121 tatgaaagaa tgcgtagaag gaaagaaaaa tgtcgtcgac atgctgaaca acaagattaa
     3181 tatgcctccg tgtataaaaa aaatattgaa cgatttgaaa gaaaacaatg taccgcgcgg
     3241 cggtatgtac aggaagaggt ttatactaaa ctgttacatt gcaaacgtgg tttcgtgtgc
     3301 caagtgtgaa aaccgatgtt taatcaaggc tctgacgcat ttctacaacc acgactccaa
     3361 gtgtgtgggt gaagtcatgc atcttttaat caaatcccaa gatgtgtata aaccaccaaa
     3421 ctgccaaaaa atgaaaactg tcgacaagct ctgtccgttt gctggcaact gcaagggtct
     3481 caatcctatt tgtaattatt gaataataaa acaattataa atgtcaaatt tgttttttat
     3541 taacgataca aaccaaacgc aacaagaaca tttgtagtat tatctataat tgaaaacgcg
     3601 tagttataat cgctgaggta atatttaaaa tcattttcaa atgattcaca gttaatttgc
     3661 gacaatataa ttttattttc acataaacta gacgccttgt cgtcttcttc ttcgtattcc
     3721 ttctcttttt catttttctc ctcataaaaa ttaacatagt tattatcgta tccatatatg
     3781 tatctatcgt atagagtaaa ttttttgttg tcataaatat atatgtcttt tttaatgggg
     3841 tgtatagtac cgctgcgcat agtttttctg taatttacaa cagtgctatt ttctggtagt
     3901 tcttcggagt gtgttgcttt aattattaaa tttatataat caatgaattt gggatcgtcg
     3961 gttttgtaca atatgttgcc ggcatagtac gcagcttctt ctagttcaat tacaccattt
     4021 tttagcagca ccggattaac ataactttcc aaaatgttgt acgaaccgtt aaacaaaaac
     4081 agttcacctc ccttttctat actattgtct gcgagcagtt gtttgttgtt aaaaataaca
     4141 gccattgtaa tgagacgcac aaactaatat cacaaactgg aaatgtctat caatatatag
     4201 ttgctgatat aacttcgtat aatgtatgct atacgaagtt atatcatgga gataattaaa
     4261 atgataacca tctcgcaaat aaataagtat tttactgttt tcgtaacagt tttgtaataa
     4321 aaaaacctat aaatacggat ccctgcagga tcaaacaagt ttgtacaaaa aagctgaacg
     4381 agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata aaaaacagac
     4441 tacataatac tgtaaaacac aacatatcca gtcactatgg cggccgcatt aggcacccca
     4501 ggctttacac tttatgcttc cggctcgtat aatgtgtgga ttttgagtta ggatccgtcg
     4561 agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt
     4621 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
     4681 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
     4741 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
     4801 gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
     4861 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
     4921 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
     4981 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg
     5041 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
     5101 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
     5161 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
     5221 tgcgatgagt ggcaggcggg gcgtaatcta gaggatccgg cttactaaaa gccagataac
     5281 agtatgcgta tttgcgcgct gatttttgcg gtataagaat atatactgat atgtataccc
     5341 gaagtatgtc aaaaagaggt atgctatgaa gcagcgtatt acagtgacag ttgacagcga
     5401 cagctatcag ttgctcaagg catatatgat gtcaatatct ccggtctggt aagcacaacc
     5461 atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg
     5521 atggctgagg tcgcccggtt tattgaaatg aacggctctt ttgctgacga gaacaggggc
     5581 tggtgaaatg cagtttaagg tttacaccta taaaagagag agccgttatc gtctgtttgt
     5641 ggatgtacag agtgatatta ttgacacgcc cgggcgacgg atggtgatcc ccctggccag
     5701 tgcacgtctg ctgtcagata aagtcccccg tgaactttac ccggtggtgc atatcgggga
     5761 tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtctccg ttatcgggga
     5821 agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta acctgatgtt
     5881 ctggggaata taaatgtcag gctcccttat acacagccag tctgcaggtc gaccatagtg
     5941 actggatatg ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta
     6001 atatattgat atttatatca ttttacgttt ctcgttcagc tttcttgtac aaagtggtag
     6061 gatccgaaca aaaactcatc tcagaagagg atctgtaagc ttgatgggcg gccgcttaat
     6121 taattgatcc gggttattag tacatttatt aagcgctaga ttctgtgcgt tgttgattta
     6181 cagacaattg ttgtacgtat tttaataatt cattaaattt ataatcttta gggtggtatg
     6241 ttagagcgaa aatcaaatga ttttcagcgt ctttatatct gaatttaaat attaaatcct
     6301 caatagattt gtaaaatagg tttcgattag tttcaaacaa gggttgtttt tccgaaccga
     6361 tggctggact atctaatgga ttttcgctca acgccacaaa acttgccaaa tcttgtagca
     6421 gcaatctagc tttgtcgata ttcgtttgtg ttttgttttg taataaaggt tcgacgtcgt
     6481 tcaaaatatt atgcgctttt gtatttcttt catcactgtc gttagtgtac aattgactcg
     6541 acgtaaacac gttaaataaa gcttggacat atttaacatc gggcgtgtta gctttattag
     6601 gccgattatc gtcgtcgtcc caaccctcgt cgttagaagt tgcttccgaa gacgattttg
     6661 ccatagccac acgacgccta ttaattgtgt cggctaacac gtccgcgatc aaatttgtag
     6721 ttgagctttt tggaattatt tctgattgcg ggcgtttttg ggcgggtttc aatctaactg
     6781 tgcccgattt taattcagac aacacgttag aaagcgatgg tgcaggcggt ggtaacattt
     6841 cagacggcaa atctactaat ggcggcggtg gtggagctga tgataaatct accatcggtg
     6901 gaggcgcagg cggggctggc ggcggaggcg gaggcggagg tggtggcggt gatgcagacg
     6961 gcggtttagg ctcaaatgtc tctttaggca acacagtcgg cacctcaact attgtactgg
     7021 tttcgggcgc cgtttttggt ttgaccggtc tgagacgagt gcgatttttt tcgtttctaa
     7081 tagcttccaa caattgttgt ctgtcgtcta aaggtgcagc gggttgaggt tccgtcggca
     7141 ttggtggagc gggcggcaat tcagacatcg atggtggtgg tggtggtgga ggcgctggaa
     7201 tgttaggcac gggagaaggt ggtggcggcg gtgccgccgg tataatttgt tctggtttag
     7261 tttgttcgcg cacgattgtg ggc
//
