LOCUS       sequence-466896-        6527 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 6527)
  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 6527)
  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: pJC175e
FEATURES             Location/Qualifiers
     source          1..6527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..951)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      999..1221
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             1979..3253
                     /codon_start=1
                     /gene="III"
                     /product="pIII"
                     /label=M13 gene III
                     /note="minor coat protein"
                     /translation="MKKLLFAIPLVVPFYSHSAETVESCLAKPHTENSFTNVWKDDKTL
                     DRYANYEGCLWNATGVVVCTGDETQCYGTWVPIGLAIPENEGGGSEGGGSEGGGSEGGG
                     TKPPEYGDTPIPGYTYINPLDGTYPPGTEQNPANPNPSLEESQPLNTFMFQNNRFRNRQ
                     GALTVYTGTVTQGTDPVKTYYQYTPVSSKAMYDAYWNGKFRDCAFHSGFNEDPFVCEYQ
                     GQSSDLPQPPVNAGGGSGGGSGGGSEGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKM
                     ANANKGAMTENADENALQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGS
                     NSQMAQVGDGDNSPLMNNFRQYLPSLPQSVECRPFVFGAGKPYEFSIDCDKINLFRGVF
                     AFLLYVATFMYVFSTFANILRNKES"
     CDS             3253..4332
                     /codon_start=1
                     /gene="luxA"
                     /product="LuxA luciferase subunit"
                     /label=LuxA
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
                     FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
                     ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
                     SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
                     DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
                     LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
                     KLFQSDVMPFLKEKQRSLLY"
     CDS             4396..5376
                     /codon_start=1
                     /gene="luxB"
                     /product="LuxB luciferase subunit"
                     /label=LuxB
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="KFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENHF
                     SDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVRIAEEACLLDQLSEGRFILGF
                     SDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAYT
                     PGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEID
                     HQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECITA
                     AKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHTEYT"
     CDS             complement(join(5456..6247,6248..6316))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6079..6098
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6317..6421)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6489..6507
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag
       61 ccatgagaac gaaccattga gatcatgctt actttgcatg tcactcaaaa attttgcctc
      121 aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt
      181 acgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg
      241 gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa
      301 cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa
      361 atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt
      421 ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt
      481 tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc
      541 aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg
      601 caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt
      661 ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat
      721 cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat
      781 ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat
      841 cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc
      901 atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa
      961 ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat gataattact
     1021 agtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact
     1081 tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta
     1141 tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga
     1201 tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc
     1261 gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac
     1321 cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga
     1381 gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg
     1441 gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag
     1501 aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct
     1561 gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta
     1621 tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca
     1681 ggcttacccg tcttactgtc aagaggacat ccggtcaaat aaaacgaaag gctcagtcga
     1741 aagactgggc ctttcgtttt gctgaggaga cttagggacc ctgatgaaat tcgccacttg
     1801 ttagtgtaat tcgctaactc atcctggcat gttgctgttg attcttcaat cagatcttta
     1861 taaatcaaaa agataaaaaa ttggcacgca aattgtatta acagttcagc aggacaatcc
     1921 tgaacgcaga aatcaagagg acaacattac ctgcaggtgc agtaaggagg aaaaaaaaat
     1981 gaaaaaatta ttattcgcaa ttcctttagt tgttcctttc tattctcact ccgctgaaac
     2041 tgttgaaagt tgtttagcaa aaccccatac agaaaattca tttactaacg tctggaaaga
     2101 cgacaaaact ttagatcgtt acgctaacta tgagggctgt ctgtggaatg ctacaggcgt
     2161 tgtagtttgt actggtgacg aaactcagtg ttacggtaca tgggttccta ttgggcttgc
     2221 tatccctgaa aatgagggtg gtggctctga gggtggcggt tctgagggtg gcggttctga
     2281 gggtggcggt actaaacctc ctgagtacgg tgatacacct attccgggct atacttatat
     2341 caaccctctc gacggcactt atccgcctgg tactgagcaa aaccccgcta atcctaatcc
     2401 ttctcttgag gagtctcagc ctcttaatac tttcatgttt cagaataata ggttccgaaa
     2461 taggcagggg gcattaactg tttatacggg cactgttact caaggcactg accccgttaa
     2521 aacttattac cagtacactc ctgtatcatc aaaagccatg tatgacgctt actggaacgg
     2581 taaattcaga gactgcgctt tccattctgg ctttaatgag gatccattcg tttgtgaata
     2641 tcaaggccaa tcgtctgacc tgcctcaacc tcctgtcaat gctggcggcg gctctggtgg
     2701 tggttctggt ggcggctctg agggtggtgg ctctgagggt ggcggttctg agggtggcgg
     2761 ctctgaggga ggcggttccg gtggtggctc tggttccggt gattttgatt atgaaaagat
     2821 ggcaaacgct aataaggggg ctatgaccga aaatgccgat gaaaacgcgc tacagtctga
     2881 cgctaaaggc aaacttgatt ctgtcgctac tgattacggt gctgctatcg atggtttcat
     2941 tggtgacgtt tccggccttg ctaatggtaa tggtgctact ggtgattttg ctggctctaa
     3001 ttcccaaatg gctcaagtcg gtgacggtga taattcacct ttaatgaata atttccgtca
     3061 atatttacct tccctccctc aatcggttga atgtcgccct tttgtctttg gcgctggtaa
     3121 accttacgag ttcagtatcg actgcgataa gatcaacctg ttccgcggtg tctttgcgtt
     3181 tcttttatat gttgccacct ttatgtatgt attttctacg tttgctaaca tactgcgtaa
     3241 taaggagtct taatgaaatt tggaaacttt ttgcttacat accaacctcc ccaattttcc
     3301 caaacagagg taatgaaacg tttggttaaa ttaggtcgca tctctgagga gtgtggtttt
     3361 gataccgtat ggttactgga gcatcatttc acggagtttg gtttgcttgg taacccttat
     3421 gtcgctgctg catatttact tggcgcgact aaaaaattga atgtaggaac tgccgctatt
     3481 gttcttccca cagcccatcc agtacgccaa cttgaagatg tgaatttatt ggatcaaatg
     3541 tcaaaaggac gatttcggtt tggtatttgc cgagggcttt acaacaagga ctttcgcgta
     3601 ttcggcacag atatgaataa cagtcgcgcc ttagcggaat gctggtacgg gctgataaag
     3661 aatggcatga cagagggata tatggaagct gataatgaac atatcaagtt ccataaggta
     3721 aaagtaaacc ccgcggcgta tagcagaggt ggcgcaccgg tttatgtggt ggctgaatca
     3781 gcttcgacga ctgagtgggc tgctcaattt ggcctaccga tgatattaag ttggattata
     3841 aatactaacg aaaagaaagc acaacttgag ctttataatg aagtggctca agaatatggg
     3901 cacgatattc ataatatcga ccattgctta tcatatataa catctgtaga tcatgactca
     3961 attaaagcga aagagatttg ccggaaattt ctggggcatt ggtatgattc ttatgtgaat
     4021 gctacgacta tttttgatga ttcagaccaa acaagaggtt atgatttcaa taaagggcag
     4081 tggcgtgact ttgtattaaa aggacataaa gatactaatc gccgtattga ttacagttac
     4141 gaaatcaatc ccgtgggaac gccgcaggaa tgtattgaca taattcaaaa agacattgat
     4201 gctacaggaa tatcaaatat ttgttgtgga tttgaagcta atggaacagt agacgaaatt
     4261 attgcttcca tgaagctctt ccagtctgat gtcatgccat ttcttaaaga aaaacaacgt
     4321 tcgctattat attatggcgg tggcggtagc ggcggtggcg gtagcggcgg tggcggtagc
     4381 ggcggtggcg gtagcaaatt tggattgttc ttccttaact tcatcaattc aacaactgtt
     4441 caagaacaga gtatagttcg catgcaggaa ataacggagt atgttgataa gttgaatttt
     4501 gaacagattt tagtgtatga aaatcatttt tcagataatg gtgttgtcgg cgctcctctg
     4561 actgtttctg gttttctgct cggtttaaca gagaaaatta aaattggttc attaaatcac
     4621 atcattacaa ctcatcatcc tgtccgcata gcggaggaag cttgcttatt ggatcagtta
     4681 agtgaaggga gatttatttt agggtttagt gattgcgaaa aaaaagatga aatgcatttt
     4741 tttaatcgcc cggttgaata tcaacagcaa ctatttgaag agtgttatga aatcattaac
     4801 gatgctttaa caacaggcta ttgtaatcca gataacgatt tttatagctt ccctaaaata
     4861 tctgtaaatc cccatgctta tacgccaggc ggacctcgga aatatgtaac agcaaccagt
     4921 catcatattg ttgagtgggc ggccaaaaaa ggtattcctc tcatctttaa gtgggatgat
     4981 tctaatgatg ttagatatga atatgctgaa agatataaag ccgttgcgga taaatatgac
     5041 gttgacctat cagagataga ccatcagtta atgatattag ttaactataa cgaagatagt
     5101 aataaagcta aacaagagac gcgtgcattt attagtgatt atgttcttga aatgcaccct
     5161 aatgaaaatt tcgaaaataa acttgaagaa ataattgcag aaaacgctgt cggaaattat
     5221 acggagtgta taactgcggc taagttggca attgaaaagt gtggtgcgaa aagtgtattg
     5281 ctgtcctttg aaccaatgaa tgatttgatg agccaaaaaa atgtaatcaa tattgttgat
     5341 gataatatta agaagtacca cacggaatat acctaaactt aattaacggc actcctcagc
     5401 aaatataatg accctcttga taacccaaga gggcattttt taatgcccat ggcgtttacc
     5461 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5521 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5581 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5641 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5701 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5761 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5821 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5881 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5941 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6001 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6061 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6121 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6181 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6241 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6301 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6361 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6421 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     6481 cctataaaaa taggcgtatc acgaggccct taggccaaat aggccgt
//
