LOCUS       Exported                3673 bp ds-DNA     circular SYN 23-OCT-2020
DEFINITION  RBS-His6-Halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3673)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Oct 30, 2020 from SnapGene Viewer 5.0.8
            https://www.snapgene.com
COMMENT     Insert Molecule:   C514-P54, 1046 bps DNA (attB5r, attB1r sites)
            Donor Vector:   pDonr-235, 4749 bps DNA circular (attP5r, attP1r 
            sites)
            Recombination using BP clonase to produce Entry Clone
FEATURES             Location/Qualifiers
     source          1..3673
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    complement(43..166)
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             202..219
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis-
                     /translation="HHHHHH"
     CDS             220..1110
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)-
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             1123..1143
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     protein_bind    1159..1283
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1356..1374)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1379..1395)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1820..2611
                     /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      2704..3292
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      3427..3454
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      3546..3632
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agcgaattct acaggtcact aataccatct aagtagttga ttcatagtga ctgcatatgt
       61 tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata
      121 tttatatcat tttacgtttc tcgttcaact tttgtataca aagtggtcat ttaactttaa
      181 gaaggagata tacatatggg ccaccatcac catcaccata tggcagaaat cggtactggc
      241 tttccattcg acccccatta tgtggaagtc ctgggcgagc gcatgcacta cgtcgatgtt
      301 ggtccgcgcg atggcacccc tgtgctgttc ctgcacggta acccgacctc ctcctacgtg
      361 tggcgcaaca tcatcccgca tgttgcaccg acccatcgct gcattgctcc agacctgatc
      421 ggtatgggca aatccgacaa accagacctg ggttatttct tcgacgacca cgtccgcttc
      481 atggatgcct tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat tcacgactgg
      541 ggctccgctc tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa aggtattgca
      601 tttatggagt tcatccgccc tatcccgacc tgggacgaat ggccagaatt tgcccgcgag
      661 accttccagg ccttccgcac caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt
      721 tttatcgagg gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt cgagatggac
      781 cattaccgcg agccgttcct gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac
      841 gagctgccaa tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga atacatggac
      901 tggctgcacc agtcccctgt cccgaagctg ctgttctggg gcaccccagg cgttctgatc
      961 ccaccggccg aagccgctcg cctggccaaa agcctgccta actgcaaggc tgtggacatc
     1021 ggcccgggtc tgaatctgct gcaagaagac aacccggacc tgatcggcag cgagatcgcg
     1081 cgctggctgt cgacgctcga gatttccggc gagccaacca ctgaggatct gtactttcag
     1141 agcgataacg cgatcgccac aactttgtac aaaaaagttg aacgagaaac gtaaaatgat
     1201 ataaatatca atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa
     1261 acacaacata tgcagtcact atgaaccaac tacttagatg gtattagtga cctgtagaat
     1321 tcgagctcta gagctgcagg gcggccgcga tatcccctat agtgagtcgt attacatggt
     1381 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     1441 agatcggtac ccggggatcc tctagaccag ccaggacaga aatgcctcga cttcgctgct
     1501 acccaaggtt gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa cccagtggac
     1561 ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac gcaactggtc
     1621 cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta
     1681 tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc
     1741 cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc
     1801 taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc
     1861 agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta
     1921 cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc tggttacggt
     1981 gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc
     2041 ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga
     2101 cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg
     2161 caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt
     2221 gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt
     2281 tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa
     2341 ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg
     2401 gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga
     2461 gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca
     2521 agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa
     2581 aggcgagatc accaaggtag tcggcaaata accctcgagc cacccatgac caaaatccct
     2641 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2701 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2761 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2821 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2881 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2941 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3001 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3061 gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca cgcttcccga
     3121 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3181 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3241 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3301 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3361 tgcgttatcc cctgattctg tggataaccg tattaccgct agccaggaag agtttgtaga
     3421 aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat
     3481 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc
     3541 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
     3601 cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt
     3661 aacgctacca tgg
//
