LOCUS       sequence-148320-        8085 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 8085)
  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 8085)
  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: pAB076i5
FEATURES             Location/Qualifiers
     source          1..8085
                     /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             3207..4286
                     /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             4350..5330
                     /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(5738..5761)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      complement(6593..6620)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6712..6798)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     6951..6968
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(join(7014..7805,7806..7874))
                     /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     7637..7656
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7875..7979)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8047..8065
                     /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 taattttaat 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 ctgcttttcc agtctgacca cttcggctta
     1621 tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggg ctcatcaaca
     1681 ggcttaccgg tcgtacggtc aagaggacat ccgggcaaat gcaacgaaag gctcagtcga
     1741 aagactgggc ctttcgttgc gctgcggaga cttagggacc ctaacacaag aaaaactgta
     1801 tttgacaaac aagatacatt gtatgaaaat acaagaaagt ttgttcatta ggcaccccgg
     1861 gctttactcg taaagcttcc ggcgcgtatg ttgtgtcgac cgacctgcag gtgcagtaag
     1921 gaggaaaaaa aaatgaaaaa attattattc gcaattcctt tagttgttcc tttctattct
     1981 cactccgctg aaactgttga aagttgttta gcaaaacccc atacagaaaa ttcatttact
     2041 aacgtctgga aagacgacaa aactttagat cgttacgcta actatgaggg ctgtctgtgg
     2101 aatgctacag gcgttgtagt ttgtactggt gacgaaactc agtgttacgg tacatgggtt
     2161 cctattgggc ttgctatccc tgaaaatgag ggtggtggct ctgagggtgg tggttctgag
     2221 ggtggcggtt ctgagggtgg cggtactaaa cctcctgagt acggtgatac acctattccg
     2281 ggctatactt atatcaaccc tctcgacggc acttatccgc ctggtactga gcaaaacccc
     2341 gctaatccta atccttctct tgaggagtct cagcctctta atactttcat gtttcagaat
     2401 aataggttcc gaaataggca gggggcatta actgtttata cgggcactgt tactcaaggc
     2461 actgaccccg ttaaaactta ttaccagtac actcctgtat catcaaaagc catgtatgac
     2521 gcttactgga acggtaaatt cagagactgc gctttccatt ctggctttaa tgaggatcca
     2581 ttcgtttgtg aatatcaagg ccaatcgtct gacctgcctc aacctcctgt caatgctggc
     2641 ggcggctctg gtggtggttc tggtggcggc tctgagggtg gtggctctga gggtggcggt
     2701 tctgagggtg gcggctctga gggaggcggt tccggtggtg gctctggttc cggtgatttt
     2761 gattatgaaa agatggcaaa cgctaataag ggggctatga ccgaaaatgc cgatgaaaac
     2821 gcgctacagt ctgacgctaa aggcaaactt gattctgtcg ctactgatta cggtgctgct
     2881 atcgatggtt tcattggtga cgtttccggc cttgctaatg gtaatggtgc tactggtgat
     2941 tttgctggct ctaattccca aatggctcaa gtcggtgacg gtgataattc acctttaatg
     3001 aataatttcc gtcaatattt accttccctc cctcaatcgg ttgaatgtcg cccttttgtc
     3061 tttggcgctg gtaaacctta cgagttcagt atcgactgcg ataagatcaa cctgttccgc
     3121 ggtgtctttg cgtttctttt atatgttgcc acctttatgt atgtattttc tacgtttgct
     3181 aacatactgc gtaataagga gtcttaatga aatttggaaa ctttttgctt acataccaac
     3241 ctccccaatt ttcccaaaca gaggtaatga aacgtttggt taaattaggt cgcatctctg
     3301 aggagtgtgg ttttgatacc gtatggttac tggagcatca tttcacggag tttggtttgc
     3361 ttggtaaccc ttatgtcgct gctgcatatt tacttggcgc gactaaaaaa ttgaatgtag
     3421 gaactgccgc tattgttctt cccacagccc atccagtacg ccaacttgaa gatgtgaatt
     3481 tattggatca aatgtcaaaa ggacgatttc ggtttggtat ttgccgaggg ctttacaaca
     3541 aggactttcg cgtattcggc acagatatga ataacagtcg cgccttagcg gaatgctggt
     3601 acgggctgat aaagaatggc atgacagagg gatatatgga agctgataat gaacatatca
     3661 agttccataa ggtaaaagta aaccccgcgg cgtatagcag aggtggcgca ccggtttatg
     3721 tggtggctga atcagcttcg acgactgagt gggctgctca atttggccta ccgatgatat
     3781 taagttggat tataaatact aacgaaaaga aagcacaact tgagctttat aatgaagtgg
     3841 ctcaagaata tgggcacgat attcataata tcgaccattg cttatcatat ataacatctg
     3901 tagatcatga ctcaattaaa gcgaaagaga tttgccggaa atttctgggg cattggtatg
     3961 attcttatgt gaatgctacg actatttttg atgattcaga ccaaacaaga ggttatgatt
     4021 tcaataaagg gcagtggcgt gactttgtat taaaaggaca taaagatact aatcgccgta
     4081 ttgattacag ttacgaaatc aatcccgtgg gaacgccgca ggaatgtatt gacataattc
     4141 aaaaagacat tgatgctaca ggaatatcaa atatttgttg tggatttgaa gctaatggaa
     4201 cagtagacga aattattgct tccatgaagc tcttccagtc tgatgtcatg ccatttctta
     4261 aagaaaaaca acgttcgcta ttatattatg gcggtggcgg tagcggcggt ggcggtagcg
     4321 gcggtggcgg tagcggcggt ggcggtagca aatttggatt gttcttcctt aacttcatca
     4381 attcaacaac tgttcaagaa cagagtatag ttcgcatgca ggaaataacg gagtatgttg
     4441 ataagttgaa ttttgaacag attttagtgt atgaaaatca tttttcagat aatggtgttg
     4501 tcggcgctcc tctgactgtt tctggttttc tgctcggttt aacagagaaa attaaaattg
     4561 gttcattaaa tcacatcatt acaactcatc atcctgtccg catagcggag gaagcttgct
     4621 tattggatca gttaagtgaa gggagattta ttttagggtt tagtgattgc gaaaaaaaag
     4681 atgaaatgca tttttttaat cgcccggttg aatatcaaca gcaactattt gaagagtgtt
     4741 atgaaatcat taacgatgct ttaacaacag gctattgtaa tccagataac gatttttata
     4801 gcttccctaa aatatctgta aatccccatg cttatacgcc aggcggacct cggaaatatg
     4861 taacagcaac cagtcatcat attgttgagt gggcggccaa aaaaggtatt cctctcatct
     4921 ttaagtggga tgattctaat gatgttagat atgaatatgc tgaaagatat aaagccgttg
     4981 cggataaata tgacgttgac ctatcagaga tagaccatca gttaatgata ttagttaact
     5041 ataacgaaga tagtaataaa gctaaacaag agacgcgtgc atttattagt gattatgttc
     5101 ttgaaatgca ccctaatgaa aatttcgaaa ataaacttga agaaataatt gcagaaaacg
     5161 ctgtcggaaa ttatacggag tgtataactg cggctaagtt ggcaattgaa aagtgtggtg
     5221 cgaaaagtgt attgctgtcc tttgaaccaa tgaatgattt gatgagccaa aaaaatgtaa
     5281 tcaatattgt tgatgataat attaagaagt accacacgga atatacctaa acggagccaa
     5341 tgtacgcaaa aaaccccgct tcagcggggt tttttcgcca aaaaaaaccc cgccctgtca
     5401 ggggcggggt ttttttttag ggccctactg actgttagcg tttcggcgcc ggataatgca
     5461 gcgtggtgat cagaccatcc gcaaccgtgc tatggtgatg caccagttct gccagggtat
     5521 tgaagcgaga ttcactagaa acgtacagtt tgccatcgct ggcggtgttg atacggtagt
     5581 gatacacgcg accttcataa cgcagagaaa tgctgcgctg gcccgggcta ctttcacttt
     5641 cacgaaccag aaaagaacca ttaatgccag agctcagcag gtattcggcc gcgttacggc
     5701 tcacagggcc atgataccag ctgtgttttt ccagactctt gtcatcgtcg tccttgtagt
     5761 ctacgaattt taccctcgct tccacgacaa caccgataat cttgcagttt ccgttgatag
     5821 gagtcatagg ccatgaagga ttcaggcctt tcaggtactt ctgaccgcca tctatgacca
     5881 gtttcttgaa tgttgcttcg ttcgcgtcag tcagtttggc tacaacaagg cttccattca
     5941 ctggctcccg tccagtatct actaacacca tatgaccttc agggatgctt tgacctacag
     6001 gtgaggtcat ggaatcacct tcaaccttca gccagaatcc attgcctaat aagttaacgt
     6061 cactgtcata ccattcatca atgtccttga tatcgtaggg ttcacaagct tcacaccacg
     6121 aacgagctct aaccatgcta atcaatggat atttcccttt gggctcaacg tgcccaacaa
     6181 atctaacatt cgaatcagag gtgccattga gcagccagtc aacacttacg ccaagagctg
     6241 acgcaagttc tggtaaaaag cgtggtcgct tagttttacc gttttcgagc tgctctatag
     6301 actgctgggt agtccccacc ttttgagcaa gttcagcctg gttaagtcca agctgaattc
     6361 ttttgctttt taccctgctg ctgatactca tttttttttc ctccttactc tagtaaaagt
     6421 taaacaaaat tatttgtaga gggaaaccgt tgtggtctcc ctgaatatag ataccgagcc
     6481 ttatgcatgc ccgtaaagtt atccagcaac cactcataga cctagggcag cagataggga
     6541 cgacgtggtg ttagctgtgc tctagaatcc tcagcgagtt tgtagaaacg caaaaaggcc
     6601 atccgtcagg atggccttct gcttaatttg atgcctggca gtttatggcg ggcgtcctgc
     6661 ccgccaccct ccgggccgtt gcttcgcaac gttcaaatcc gctcccggcg gatttgtcct
     6721 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg
     6781 agcctttcgt tttatttgat gcctggcagt tccctactct cgcatgggga gaccccacac
     6841 taccatcggc gctacggcgt ttcacttctg agttcggcat ggggtcaggt gggaccaccg
     6901 cgctactgcc gccaggcaaa ttctgtttta tcagaccgct tctgcgttct gatttaatct
     6961 gtatcaggct gaaaatcttc tctcatccgc caaaacagcc atgcccatgg cgtttaccaa
     7021 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7081 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7141 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7201 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7261 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7321 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7381 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7441 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7501 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7561 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7621 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7681 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7741 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7801 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7861 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7921 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7981 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     8041 tataaaaata ggcgtatcac gaggccctta ggccaaatag gccgt
//
