LOCUS       sequence-330540-        3081 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4/attB1r entry clone for baculovirus expression of His6 
            tagged proteins using the polyhedrin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3081)
  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 3081)
  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-E01: attB4-polyhedrinp>His6-nostop-attB1r
FEATURES             Location/Qualifiers
     source          1..3081
                     /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"
     protein_bind    387..511
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(583..602)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(584..602)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(607..623)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(607..623)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1048..1839
                     /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      1932..2520
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2421..2440
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2674..2691
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2850..2877)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2969..3055)
                     /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 caccatacaa ctttgtacaa aaaagttgaa cgagaaacgt
      421 aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag actacataat
      481 actgtaaaac acaacatatg cagtcactat gaaccaacta cttagatggt attagtgacc
      541 tgtagaattc gagctctaga gctgcagggc ggccgcgata tcccctatag tgagtcgtat
      601 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
      661 attgcacaag atcggtaccc ggggatcctc tagaccagcc aggacagaaa tgcctcgact
      721 tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc
      781 cagtggacat aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc
      841 aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg
      901 gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg
      961 cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc
     1021 agtcgcccta aaacaaagtt aaacattatg agggaagcgg tgatcgccga agtatcgact
     1081 caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta
     1141 catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg
     1201 gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg
     1261 gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt
     1321 gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa
     1381 tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg
     1441 gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag
     1501 gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg
     1561 ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc
     1621 cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg
     1681 gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat
     1741 cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac
     1801 tacgtgaaag gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca
     1861 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     1921 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1981 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2041 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     2101 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2161 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2221 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     2281 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     2341 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2401 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2461 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     2521 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     2581 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     2641 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     2701 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     2761 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
     2821 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
     2881 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
     2941 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
     3001 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
     3061 tggcagttcc ctactctcgc g
//
