LOCUS       sequence-330534-        3057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4/attB1r entry clone for baculovirus expression using 
            the AcMNPV polyhedrin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3057)
  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 3057)
  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: C413-E22: attB4-polyhedrinp>-attB1r
FEATURES             Location/Qualifiers
     source          1..3057
                     /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"
     protein_bind    363..487
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(559..578)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(560..578)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(583..599)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(583..599)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1024..1815
                     /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      1908..2496
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2397..2416
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2650..2667
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2826..2853)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2945..3031)
                     /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 atacaacttt gtacaaaaaa gttgaacgag aaacgtaaaa tgatataaat atcaatatat
      421 taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa catatgcagt
      481 cactatgaac caactactta gatggtatta gtgacctgta gaattcgagc tctagagctg
      541 cagggcggcc gcgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg
      601 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagatcg gtacccgggg
      661 atcctctaga ccagccagga cagaaatgcc tcgacttcgc tgctacccaa ggttgccggg
      721 tgacgcacac cgtggaaacg gatgaaggca cgaacccagt ggacataagc ctgttcggtt
      781 cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa
      841 cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg tttttttggg
      901 gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg
      961 atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac aaagttaaac
     1021 attatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt agttggcgtc
     1081 atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc cgcagtggat
     1141 ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt aaggcttgat
     1201 gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc ccctggagag
     1261 agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat cattccgtgg
     1321 cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga cattcttgca
     1381 ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac aaaagcaaga
     1441 gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc ggttcctgaa
     1501 caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg
     1561 gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag cgcagtaacc
     1621 ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct gccggcccag
     1681 tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga agatcgcttg
     1741 gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga gatcaccaag
     1801 gtagtcggca aataaccctc gagccaccca tgaccaaaat cccttaacgt gagttacgcg
     1861 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     1921 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     1981 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2041 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2101 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2161 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2221 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2281 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2341 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2401 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     2461 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     2521 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     2581 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     2641 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     2701 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     2761 gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa
     2821 acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg
     2881 gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg
     2941 gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc
     3001 agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcg
//
