LOCUS       sequence-148365-        4198 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Directed Evolution of Protein-Protein Interactions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4198)
  AUTHORS   Badran AH, Guzov VM, Huai Q, Kemp MM, Vishwanath P, Kain W, Nance 
            AM, Evdokimov A, Moshiri F, Turner KH, Wang P, Malvar T, Liu DR
  TITLE     Continuous evolution of Bacillus thuringiensis toxins overcomes 
            insect resistance.
  JOURNAL   Nature. 2016 May 5;533(7601):58-63. doi: 10.1038/nature17938. Epub 
            2016 Apr 27.
  PUBMED    27120167
REFERENCE   2  (bases 1 to 4198)
  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"; doi: 
            "10.1038/nature17938"; journalName: "Nature"; date: "2016-05-5- 5"; 
            volume: "533"; issue: "7601"; pages: "58-63"
COMMENT     Sequence Label: pAB094d
FEATURES             Location/Qualifiers
     source          1..4198
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     489..508
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     742..759
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    774..914
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(840..859)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1000..1022
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1016..1207)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      1318..1363
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1378..2256)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2283..2567
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     2490..2509
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2610..2882
                     /codon_start=1
                     /gene="E. coli rpoZ"
                     /product="DNA-directed RNA polymerase subunit omega"
                     /label=RNAP omega
                     /note="promotes RNA polymerase assembly"
                     /translation="MARVTVQDAVEKIGNRFDLVLVAARRARQMQVGGKDPLVPEENDK
                     TTVIALREIEEGLINNQILDVRERQEQQEQEAAELQAVTAIAEGRR"
     CDS             2892..2921
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3306..4097)
                     /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"
     promoter        complement(4098..4189)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(4198,1..588)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
       61 gcggtggttt gtttgccgga tcaagagcta ccacctcttt ttccgaaggt aactggcttc
      121 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
      181 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
      241 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
      301 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
      361 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
      421 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
      481 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
      541 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
      601 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
      661 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
      721 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
      781 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
      841 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
      901 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
      961 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     1021 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     1081 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     1141 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     1201 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     1261 aacgagagag gatgctcacg atacgggtta ctgatgatga acaagaggac atccggtcaa
     1321 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttagacttag ggacccttta
     1381 tgacaacttg acggctacat cattcacttt ttcttcacaa ccggcacgga actcgctcgg
     1441 gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt caaaaccaac
     1501 attgcgaccg acggtggcga taggcatccg ggtggtgctc aaaagcagct tcgcctggct
     1561 gatacgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc ggaaaagatg
     1621 tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat caaaattgct
     1681 gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat ccatcggtgg
     1741 atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa gcagatttat
     1801 cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt gcccaaacag
     1861 gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat tggcaaatat
     1921 tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa cccactggtg
     1981 ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg gcgggaacag
     2041 caaaatatca cccggtcggc aaacaaattc tcgtccctga tttttcacca ccccctgacc
     2101 gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga taaaaaaatc
     2161 gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat taaacgagta
     2221 tcccggcagc aggggatcat tttgcgcttc agccatactt ttcatactcc caccattcag
     2281 agaagaaacc aattgtccat attgcatcag acattgccgt cactgcgtct tttactggct
     2341 cttctcgcta acccaaccgg taaccccgct tattaaaagc attctgtaac aaagcgggac
     2401 caaagccatg acaaaaacgc gtaacaaaag tgtctataat cacggcagaa aagtccacat
     2461 tgattatttg cacggcgtca cactttgcta tgccatagca tttttatcca taagattagc
     2521 ggatcctacc tgacgctttt tatcgcaact ctctactgtt tctccatacc cgttttttta
     2581 cctgcaggtg cagtaaggag gaaaaaaaaa tggcacgcgt aactgttcag gacgctgtag
     2641 agaaaattgg taaccgtttt gacctggtac tggtcgccgc gcgtcgcgct cgtcagatgc
     2701 aggtaggcgg aaaggatccg ctggtaccgg aagaaaacga taaaaccact gtaatcgcgc
     2761 tgcgcgaaat cgaagaaggt ctgatcaaca accagatcct cgacgttcgc gaacgccagg
     2821 aacagcaaga gcaggaagcc gctgaattac aagccgttac cgctattgct gaaggtcgtc
     2881 gtgcggccgc ggaacaaaag cttatttctg aagaggactt gggcagctct gtgagtagcg
     2941 ttccgaccaa actggaagtg gttgcagcaa ccccgacgag cctgctgatt tcttgggatg
     3001 ccccgatgtc tagtagctct gtgtattact atcgtatcac ctacggtgaa acgggcggta
     3061 acagcccggt gcaggcgttt acggttccgt atagtagctc taccgcgacg attagtggcc
     3121 tgagcccggg tgtggattac accatcacgg tttatgcatg gggcgaagat agcgcgggtt
     3181 acatgttcat gtattctccg attagtatca attaccgcac ctgctaactt aattaacggc
     3241 actcctcagc aaatataatg accctcttga taacccaaga gggcattttt taatgcccat
     3301 ggcgtttatt tgccgactac cttggtgatc tcgcctttca cgtagtggac aaattcttcc
     3361 aactgatctg cgcgcgaggc caagcgatct tcttcttgtc caagataagc ctgtctagct
     3421 tcaagtatga cgggctgata ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca
     3481 tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact
     3541 acatttcgct catcgccagc ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt
     3601 agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga gttcctccgc cgctggacct
     3661 accaaggcaa cgctatgttc tcttgctttt gtcagcaaga tagccagatc aatgtcgatc
     3721 gtggctggct cgaagatacc tgcaagaatg tcattgcgct gccattctcc aaattgcagt
     3781 tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct
     3841 acagcgcgga gaatctcgct ctctccaggg gaagccgaag tttccaaaag gtcgttgatc
     3901 aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg taaccagcaa atcaatatca
     3961 ctgtgtggct tcaggccgcc atccactgcg gagccgtaca aatgtacggc cagcaacgtc
     4021 ggttcgagat ggcgctcgat gacgccaact acctctgata gttgagtcga tacttcggcg
     4081 atcaccgctt ccctcatact cttccttttt caatattatt gaagcattta tcagggttat
     4141 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataggccaaa taggccgt
//
