LOCUS       sequence-379000-        7283 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 GST tag 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_PH-GST-Nter-GWs-Lox (VE5586)
FEATURES             Location/Qualifiers
     source          1..7283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     4..1361
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(20..1361)
                     /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      1508..1963
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1595..1614)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1805..1826
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1980..1999)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2099..2121
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2159..2177)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2245..2349
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2350..2418,2419..3210)
                     /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(2568..2587)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3381..3969
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3870..3889
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4123..4140
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_recomb     4532..5359
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             4532..4656
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4694..5299)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     protein_bind    5363..5396
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5397..5488
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5413..5430
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     protein_bind    5509..5633
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(5658..5663,5664..5681,5682..5688)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             5742..6422
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(5808..5827)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             6742..7047
                     /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     6926..6945
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(7088..7212)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             7220..7249
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 cgggttatta gtacatttat taagcgctag attctgtgcg ttgttgattt acagacaatt
       61 gttgtacgta ttttaataat tcattaaatt tataatcttt agggtggtat gttagagcga
      121 aaatcaaatg attttcagcg tctttatatc tgaatttaaa tattaaatcc tcaatagatt
      181 tgtaaaatag gtttcgatta gtttcaaaca agggttgttt ttccgaaccg atggctggac
      241 tatctaatgg attttcgctc aacgccacaa aacttgccaa atcttgtagc agcaatctag
      301 ctttgtcgat attcgtttgt gttttgtttt gtaataaagg ttcgacgtcg ttcaaaatat
      361 tatgcgcttt tgtatttctt tcatcactgt cgttagtgta caattgactc gacgtaaaca
      421 cgttaaataa agcttggaca tatttaacat cgggcgtgtt agctttatta ggccgattat
      481 cgtcgtcgtc ccaaccctcg tcgttagaag ttgcttccga agacgatttt gccatagcca
      541 cacgacgcct attaattgtg tcggctaaca cgtccgcgat caaatttgta gttgagcttt
      601 ttggaattat ttctgattgc gggcgttttt gggcgggttt caatctaact gtgcccgatt
      661 ttaattcaga caacacgtta gaaagcgatg gtgcaggcgg tggtaacatt tcagacggca
      721 aatctactaa tggcggcggt ggtggagctg atgataaatc taccatcggt ggaggcgcag
      781 gcggggctgg cggcggaggc ggaggcggag gtggtggcgg tgatgcagac ggcggtttag
      841 gctcaaatgt ctctttaggc aacacagtcg gcacctcaac tattgtactg gtttcgggcg
      901 ccgtttttgg tttgaccggt ctgagacgag tgcgattttt ttcgtttcta atagcttcca
      961 acaattgttg tctgtcgtct aaaggtgcag cgggttgagg ttccgtcggc attggtggag
     1021 cgggcggcaa ttcagacatc gatggtggtg gtggtggtgg aggcgctgga atgttaggca
     1081 cgggagaagg tggtggcggc ggtgccgccg gtataatttg ttctggttta gtttgttcgc
     1141 gcacgattgt gggcaccggc gcaggcgccg ctggctgcac aacggaaggt cgtctgcttc
     1201 gaggcagcgc ttggggtggt ggcaattcaa tattataatt ggaatacaaa tcgtaaaaat
     1261 ctgctataag cattgtaatt tcgctatcgt ttaccgtgcc gatatttaac aaccgctcaa
     1321 tgtaagcaat tgtattgtaa agagattgtc tcaagctcgg atcgatcccg cacgccgata
     1381 acaagccttt tcatttttac tacagcattg tagtggcgag acacttcgct gtcgtcgcct
     1441 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     1501 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     1561 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     1621 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     1681 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     1741 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     1801 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt
     1861 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     1921 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     1981 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     2041 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     2101 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     2161 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     2221 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2281 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2341 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2401 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2461 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2521 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2581 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     2641 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     2701 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     2761 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     2821 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     2881 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     2941 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     3001 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3061 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3121 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3181 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3241 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3301 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3361 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3421 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3481 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     3541 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3601 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3661 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3721 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3781 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3841 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3901 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3961 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4021 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4081 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4141 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     4201 tgcagaacgg ctccgcccac tattaatgaa attaaaaatt ccaattttaa aaaacgcagc
     4261 aagagaaaca tttgtatgaa agaatgcgta gaaggaaaga aaaatgtcgt cgacatgctg
     4321 aacaacaaga ttaatatgcc tccgtgtata aaaaaaatat tgaacgattt gaaagaaaac
     4381 aatgtaccgc gcggcggtat gtacaggaag aggtttatac taaactgtta cattgcaaac
     4441 gtggtttcgt gtgccaagtg tgaaaaccga tgtttaatca aggctctgac gcatttctac
     4501 aaccacgact ccaagtgtgt gggtgaagtc atgcatcttt taatcaaatc ccaagatgtg
     4561 tataaaccac caaactgcca aaaaatgaaa actgtcgaca agctctgtcc gtttgctggc
     4621 aactgcaagg gtctcaatcc tatttgtaat tattgaataa taaaacaatt ataaatgtca
     4681 aatttgtttt ttattaacga tacaaaccaa acgcaacaag aacatttgta gtattatcta
     4741 taattgaaaa cgcgtagtta taatcgctga ggtaatattt aaaatcattt tcaaatgatt
     4801 cacagttaat ttgcgacaat ataattttat tttcacataa actagacgcc ttgtcgtctt
     4861 cttcttcgta ttccttctct ttttcatttt tctcctcata aaaattaaca tagttattat
     4921 cgtatccata tatgtatcta tcgtatagag taaatttttt gttgtcataa atatatatgt
     4981 cttttttaat ggggtgtata gtaccgctgc gcatagtttt tctgtaattt acaacagtgc
     5041 tattttctgg tagttcttcg gagtgtgttg ctttaattat taaatttata taatcaatga
     5101 atttgggatc gtcggttttg tacaatatgt tgccggcata gtacgcagct tcttctagtt
     5161 caattacacc attttttagc agcaccggat taacataact ttccaaaatg ttgtacgaac
     5221 cgttaaacaa aaacagttca cctccctttt ctatactatt gtctgcgagc agttgtttgt
     5281 tgttaaaaat aacagccatt gtaatgagac gcacaaacta atatcacaaa ctggaaatgt
     5341 ctatcaatat atagttgctg atataacttc gtataatgta tgctatacga agttatatca
     5401 tggagataat taaaatgata accatctcgc aaataaataa gtattttact gttttcgtaa
     5461 cagttttgta ataaaaaaac ctataaatac ggatccctgc aggatcaaac aagtttgtac
     5521 aaaaaagctg aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg
     5581 cataaaaaac agactacata atactgtaaa acacaacata tccagtcact atggcggccg
     5641 cattaggcac cccaggcttt acactttatg cttccggctc gtataatgtg tggattttga
     5701 gttaggatcc gtcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
     5761 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
     5821 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
     5881 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
     5941 tgaatgctca tccggaattc cgtatggcaa tgaaagacgg tgagctggtg atatgggata
     6001 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga
     6061 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt
     6121 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag
     6181 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct
     6241 tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc
     6301 tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg
     6361 aattacaaca gtactgcgat gagtggcagg cggggcgtaa tctagaggat ccggcttact
     6421 aaaagccaga taacagtatg cgtatttgcg cgctgatttt tgcggtataa gaatatatac
     6481 tgatatgtat acccgaagta tgtcaaaaag aggtatgcta tgaagcagcg tattacagtg
     6541 acagttgaca gcgacagcta tcagttgctc aaggcatata tgatgtcaat atctccggtc
     6601 tggtaagcac aaccatgcag aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg
     6661 aaaatcagga agggatggct gaggtcgccc ggtttattga aatgaacggc tcttttgctg
     6721 acgagaacag gggctggtga aatgcagttt aaggtttaca cctataaaag agagagccgt
     6781 tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg acggatggtg
     6841 atccccctgg ccagtgcacg tctgctgtca gataaagtcc cccgtgaact ttacccggtg
     6901 gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggtc
     6961 tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc
     7021 attaacctga tgttctgggg aatataaatg tcaggctccc ttatacacag ccagtctgca
     7081 ggtcgaccat agtgactgga tatgttgtgt tttacagtat tatgtagtct gttttttatg
     7141 caaaatctaa tttaatatat tgatatttat atcattttac gtttctcgtt cagctttctt
     7201 gtacaaagtg gtaggatccg aacaaaaact catctcagaa gaggatctgt aagcttgatg
     7261 ggcggccgct taattaattg atc
//
