LOCUS       Exported                4027 bp ds-DNA     circular SYN 23-OCT-2020
DEFINITION  L4-plhp>MBP-R1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4027)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Oct 29, 2020 from SnapGene Viewer 5.0.8
            https://www.snapgene.com
COMMENT     Insert Molecule:   C417-P06, 1332 bps DNA (attB4, attB1r sites)
            Donor Vector:   pDonr-258, 4415 bps DNA circular (attP4, attP1r 
            sites)
            Recombination using BP clonase to produce Entry Clone
FEATURES             Location/Qualifiers
     source          1..4027
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     38..54
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    84..179
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        242..333
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     CDS             346..1443
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP-
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1300..1309
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    1459..1583
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1994..2785
                     /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      2878..3466
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3591..3644
                     /label=pas
                     /note=" pBR primosome ass'y site (L)"
     terminator      3796..3823
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      3915..4001
                     /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 ggtccgagga tccaaataat gattttattt tgactgatag tgacctgttc
      121 gttgcaacaa attgataagc aatgcttttt tataatgcca actttgtata gaaaagttgg
      181 ccgcccagga ctctagctat agttctagtg gttggctacg tatactccgg aatattaata
      241 gatcatggag ataattaaaa tgataaccat ctcgcaaata aataagtatt ttactgtttt
      301 cgtaacagtt ttgtaataaa aaaacctata aatatgggat ccggcaaaat cgaagaaggt
      361 aaactggtaa tctggattaa cggcgataaa ggctataacg gtctcgctga agtcggtaag
      421 aaattcgaga aagataccgg aattaaagtc accgttgagc atccggataa actggaagag
      481 aaattcccac aggttgcggc aactggcgat ggccctgaca ttatcttctg ggcacacgac
      541 cgctttggtg gctacgctca atctggcctg ttggctgaaa tcaccccgga caaagcgttc
      601 caggacaagc tgtatccgtt tacctgggat gccgtacgtt acaacggcaa gctgattgct
      661 tacccgatcg ctgttgaagc gttatcgctg atttataaca aagatctgct gccgaacccg
      721 ccaaaaacct gggaagagat cccggcgctg gataaagaac tgaaagcgaa aggtaagagc
      781 gcgctgatgt tcaacctgca agaaccgtac ttcacctggc cgctgattgc tgctgacggg
      841 ggttatgcgt tcaagtatga aaacggcaag tacgacatta aagacgtggg cgtggataac
      901 gctggcgcga aagcgggtct gaccttcctg gttgacctga ttaaaaacaa acacatgaat
      961 gcagacaccg attactccat cgcagaagct gcctttaata aaggcgaaac agcgatgacc
     1021 atcaacggcc cgtgggcatg gtccaacatc gacaccagca aagtgaatta tggtgtaacg
     1081 gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg ttggcgtgct gagcgcaggt
     1141 attaacgccg ccagtccgaa caaagagctg gcaaaagagt tcctcgaaaa ctatctgctg
     1201 actgatgaag gtctggaagc ggttaataaa gacaaaccgc tgggtgccgt agcgctgaag
     1261 tcttacgagg aagagttggc gaaagatcca cgtattgccg ccaccatgga aaacgcccag
     1321 aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt tctggtatgc cgtgcgtact
     1381 gcggtgatca acgccgccag cggtcgtcag actgtcgatg aagccctgaa agacgcgcag
     1441 actaattcgg gatcctcgac aactttgtac aaaaaagctg aacgagaaac gtaaaatgat
     1501 ataaatatca atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa
     1561 acacaacata tgcagtcact atgaatcaac tacttagatg gtattagtga cctgtagcat
     1621 gcaagcttag atcctctaga ccagccagga cagaaatgcc tcgacttcgc tgctacccaa
     1681 ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt ggacataagc
     1741 ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac
     1801 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg
     1861 tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg
     1921 tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac
     1981 aaagttaaac attatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt
     2041 agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc
     2101 cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt
     2161 aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc
     2221 ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat
     2281 cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga
     2341 cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac
     2401 aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc
     2461 ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc
     2521 gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag
     2581 cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct
     2641 gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga
     2701 agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga
     2761 gatcaccaag gtagtcggca aataaccctc gagccaccca tgaccaaaat cccttaacgt
     2821 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2881 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2941 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3001 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     3061 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3121 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3181 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3241 caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag
     3301 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3361 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3421 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3481 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3541 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3601 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     3661 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     3721 cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga
     3781 gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg
     3841 gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa
     3901 tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa
     3961 cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac
     4021 tctcgcg
//
