LOCUS       sequence-330549-        2913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB5r/attB1r entry clone containing divergent AcMNPV 
            polyhedrin and p10 promoters, for use in construction of 
            polycistronic baculovirus expression vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2913)
  AUTHORS   Wall VE, Garvey LA, Mehalko JL, Procter LV, Esposito D
  TITLE     Combinatorial assembly of clone libraries using site-specific 
            recombination.
  JOURNAL   Methods Mol Biol. 2014;1116:193-208. doi: 
            10.1007/978-1-62703-764-8_14.
  PUBMED    24395366
REFERENCE   2  (bases 1 to 2913)
  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: "Methods Mol
            Biol."; date: "2014"; volume: "1116:193-208. doi"; pages: " 
            10.1007/978-1-62703-764-8_14"
COMMENT     Sequence Label: C513-E07: attB5r-<p10p-polyhedrinp>-attB1r
FEATURES             Location/Qualifiers
     source          1..2913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(44..167)
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(174..283)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        307..398
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     323..340
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     protein_bind    399..523
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(595..614)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(596..614)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(619..635)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(619..635)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1060..1851
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1944..2532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2433..2452
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2667..2694)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2786..2872)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agctcgaatt ctacagtcac taataccatc taagtagttg attcatagtg actgcatatg
       61 ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat
      121 atttatatca ttttacgttt ctcgttcaac ttttgtatac aaagtggtcg agtgattgta
      181 aataaaatgt aatttacagt atagtatttt aattaatata caaatgattt gataataatt
      241 cttatttaac tataatatat tgtgttgggt tgaattaaag gtccgtatac tccggaatat
      301 taatagatca tggagataat taaaatgata accatctcgc aaataaataa gtattttact
      361 gttttcgtaa cagttttgta ataaaaaaac ctataaatac aactttgtac aaaaaagttg
      421 aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg cataaaaaac
      481 agactacata atactgtaaa acacaacata tgcagtcact atgaaccaac tacttagatg
      541 gtattagtga cctgtagaat tcgagctcta gagctgcagg gcggccgcga tatcccctat
      601 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
      661 ctctgatgtt acattgcaca agatcggtac ccggggatcc tctagaccag ccaggacaga
      721 aatgcctcga cttcgctgct acccaaggtt gccgggtgac gcacaccgtg gaaacggatg
      781 aaggcacgaa cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt
      841 atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg
      901 gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc
      961 aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt
     1021 acgcagcagg gcagtcgccc taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc
     1081 gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg
     1141 ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat
     1201 attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc
     1261 aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa
     1321 gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg
     1381 caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc
     1441 gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt
     1501 ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat
     1561 gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg
     1621 cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc
     1681 gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct
     1741 agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa
     1801 gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata accctcgagc
     1861 cacccatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     1921 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1981 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2041 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     2101 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2161 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2221 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2281 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2341 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2401 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2461 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2521 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2581 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgct
     2641 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
     2701 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
     2761 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
     2821 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
     2881 cagttcccta ctctcgcgtt aacgctacca tgg
//
