LOCUS       sequence-333362-        3834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4/attB1r entry clone for transient insect expression 
            using the ie1 promoter from AcMNPV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3834)
  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 3834)
  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-E45: attB4-hr5-ie1p>-attB1r
FEATURES             Location/Qualifiers
     source          1..3834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     65..87
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     79..96
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     80..96
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    126..221
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        713..1305
                     /label=IE1 promoter
                     /note="promoter of the ie1 gene from the baculovirus 
                     Autographa californica"
     protein_bind    1308..1432
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1843..2634
                     /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      2727..3315
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3216..3235
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3469..3486
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3645..3672)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(join(3764..3834,1..16))
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg
       61 ttttcccagt cacgacgttg taaaacgacg gccagtctta aaattcgagc tcggtccgag
      121 gatccaaata atgattttat tttgactgat agtgacctgt tcgttgcaac aaattgataa
      181 gcaatgcttt tttataatgc caactttgta tagaaaagtt ggcgccgcgt aaaacacaat
      241 caagtatgag tcataagctg atgtcatgtt ttgcacacgg ctcataaccg aactggcttt
      301 acgagtagaa ttctacttgt aacgcacgat cgagtggatg atggtcattt gtttttcaaa
      361 tcgagatgat gtcatgtttt gcacacgggc tcataaactg ctttacgagt agaattctac
      421 gtgtaacgca cgatcgattg atgagtcatt tgttttgcaa tatgatatca tacaatatga
      481 ctcatttgtt tttcaaaacc gaacttgatt tacgggtaga attctactcg taaagcacaa
      541 tcaaaaagat gatgtcattt gtttttcaaa actgaactct cggctttacg agtagaattc
      601 tacgtgtaaa acacaatcaa gaaatgatgt catttgttat aaaaataaaa gctgatgtca
      661 tgttttgcac atggctcata actaaactcg ctttacgggt agaattctac gtcgatgtct
      721 ttgtgatgcg ccgacatttt tgtaggttat tgataaaatg aacggataca gttgcccgac
      781 attatcatta aatccttggc gtagaatttg tcgggtccat tgtccgtgtg cgctagcatg
      841 cccgctaacg gacctcgtac ttttggcttc aaaggttttg cgcacagaca aaatgtgcca
      901 cacttgcagc tctgcatgtg tgcgcgttac cacaaatccc aacggcgcag tgtacttgtt
      961 gtatgcaaat aaatctcgat aaaggcgcgg cgcgcgaatg cagctgatca cgtacgctcc
     1021 tcgtgttccg ttcaaggacg gtgttatcga cctcagatta atgtttatcg gccgactgtt
     1081 ttcgtatccg ctcaccaaac gcgtttttgc attaacattg tatgtcggcg gatgttctat
     1141 atctaatttg aataaataaa cgataaccgc gttggtttta gagggcataa taaaagaaat
     1201 attgttatcg tgttcgccat tagggcagta taaattgacg ttcatgttgg atattgtttc
     1261 agttgcaagt tgacactggc ggcgacaaga tcgtgaacaa ccaagtgaca actttgtaca
     1321 aaaaagctga acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc
     1381 ataaaaaaca gactacataa tactgtaaaa cacaacatat gcagtcacta tgaatcaact
     1441 acttagatgg tattagtgac ctgtagcatg caagcttaga tcctctagac cagccaggac
     1501 agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg
     1561 atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag
     1621 cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca
     1681 gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca
     1741 tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat
     1801 gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc
     1861 gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg
     1921 acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt
     1981 gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg
     2041 atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta
     2101 gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa
     2161 ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg
     2221 atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta
     2281 ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta
     2341 aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta
     2401 gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat
     2461 gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa
     2521 gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg
     2581 gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg
     2641 agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac
     2701 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2761 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2821 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     2881 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2941 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3001 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3061 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3121 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3181 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3241 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3301 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3361 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3421 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3481 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     3541 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     3601 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
     3661 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
     3721 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
     3781 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgag
//
