LOCUS       sequence-330542-        3753 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4/attB1r entry clone for baculovirus expression of 
            His6-GST tagged proteins using the polyhedrin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3753)
  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 3753)
  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: C417-E03: attB4-polyhedrinp>His6-GST-nostop-at...
FEATURES             Location/Qualifiers
     source          1..3753
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..45
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     37..54
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     38..54
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    110..205
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        271..362
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     287..304
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             369..386
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             393..1040
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="PILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFE
                     LGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGV
                     SRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYM
                     DPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     998..1020
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     protein_bind    1059..1183
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1255..1274)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1256..1274)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1279..1295)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1279..1295)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1720..2511
                     /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      2604..3192
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3093..3112
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3346..3363
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3522..3549)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3641..3727)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaaa
       61 attcgagctc ggtccgatcg ggcccgcgtt aacgctacca tggagctcca aataatgatt
      121 ttattttgac tgatagtgac ctgttcgttg caacaaattg ataagcaatg cttttttata
      181 atgccaactt tgtatagaaa agttggcgtt cgcccaggac tctagctata gttctagtgg
      241 ttggctacgt atactccgga atattaatag atcatggaga taattaaaat gataaccatc
      301 tcgcaaataa ataagtattt tactgttttc gtaacagttt tgtaataaaa aaacctataa
      361 atatgggcca ccatcaccac caccatggcg cccctatact aggttattgg aaaattaagg
      421 gccttgtgca acccactcga cttcttttgg aatatcttga agaaaaatat gaagagcatt
      481 tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg ggtttggagt
      541 ttcccaatct tccttattat attgatggtg atgttaaatt aacacagtct atggccatca
      601 tacgttatat agctgacaag cacaacatgt tgggtggttg tccaaaagag cgtgcagaga
      661 tttcaatgct tgaaggagcg gttttggata ttagatacgg tgtttcgaga attgcatata
      721 gtaaagactt tgaaactctc aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa
      781 tgttcgaaga tcgtttatgt cataaaacat atttaaatgg tgatcatgta acccatcctg
      841 acttcatgtt gtatgacgct cttgatgttg ttttatacat ggacccaatg tgcctggatg
      901 cgttcccaaa attagtttgt tttaaaaaac gtattgaagc tatcccacaa attgataagt
      961 acttgaaatc cagcaagtat atagcatggc ctttgcaggg ctggcaagcc acgtttggtg
     1021 gtggcgacca tcctccaaaa tcggatctgg ttccgcgtac aactttgtac aaaaaagttg
     1081 aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg cataaaaaac
     1141 agactacata atactgtaaa acacaacata tgcagtcact atgaaccaac tacttagatg
     1201 gtattagtga cctgtagaat tcgagctcta gagctgcagg gcggccgcga tatcccctat
     1261 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     1321 ctctgatgtt acattgcaca agatcggtac ccggggatcc tctagaccag ccaggacaga
     1381 aatgcctcga cttcgctgct acccaaggtt gccgggtgac gcacaccgtg gaaacggatg
     1441 aaggcacgaa cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt
     1501 atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg
     1561 gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc
     1621 aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt
     1681 acgcagcagg gcagtcgccc taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc
     1741 gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg
     1801 ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat
     1861 attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc
     1921 aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa
     1981 gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg
     2041 caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc
     2101 gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt
     2161 ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat
     2221 gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg
     2281 cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc
     2341 gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct
     2401 agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa
     2461 gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata accctcgagc
     2521 cacccatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     2581 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2641 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2701 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     2761 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2821 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2881 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2941 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3001 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3061 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3121 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3181 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3241 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3301 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3361 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     3421 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     3481 aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     3541 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     3601 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     3661 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
     3721 ttatttgatg cctggcagtt ccctactctc gcg
//
