LOCUS       sequence-148337-        8244 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 8244)
  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 8244)
  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: pAB088i
FEATURES             Location/Qualifiers
     source          1..8244
                     /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             3162..4241
                     /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             4305..5285
                     /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(6056..6079)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      complement(6752..6779)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6871..6957)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     7110..7127
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(join(7173..7964,7965..8033))
                     /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     7796..7815
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8034..8138)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8206..8224
                     /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 ggcttacccg tcgtacggtc aagaggacat ccgggcaaat gcaacgaaag gctcagtcga
     1741 aagactgggc ctttcgttgc gctgcggaga cttagggacc ctaatacaag aaagtttgtt
     1801 cattaggcac cccgggcttt actcgtaaag cttccggcgc gtatgttgtg tcgaccgacc
     1861 tgcaggtgca gtaaggagga aaaaaaaatg aaaaaattat tattcgcaat tcctttagtt
     1921 gttcctttct attctcactc cgctgaaact gttgaaagtt gtttagcaaa accccataca
     1981 gaaaattcat ttactaacgt ctggaaagac gacaaaactt tagatcgtta cgctaactat
     2041 gagggctgtc tgtggaatgc tacaggcgtt gtagtttgta ctggtgacga aactcagtgt
     2101 tacggtacat gggttcctat tgggcttgct atccctgaaa atgagggtgg tggctctgag
     2161 ggtggtggtt ctgagggtgg cggttctgag ggtggcggta ctaaacctcc tgagtacggt
     2221 gatacaccta ttccgggcta tacttatatc aaccctctcg acggcactta tccgcctggt
     2281 actgagcaaa accccgctaa tcctaatcct tctcttgagg agtctcagcc tcttaatact
     2341 ttcatgtttc agaataatag gttccgaaat aggcaggggg cattaactgt ttatacgggc
     2401 actgttactc aaggcactga ccccgttaaa acttattacc agtacactcc tgtatcatca
     2461 aaagccatgt atgacgctta ctggaacggt aaattcagag actgcgcttt ccattctggc
     2521 tttaatgagg atccattcgt ttgtgaatat caaggccaat cgtctgacct gcctcaacct
     2581 cctgtcaatg ctggcggcgg ctctggtggt ggttctggtg gcggctctga gggtggtggc
     2641 tctgagggtg gcggttctga gggtggcggc tctgagggag gcggttccgg tggtggctct
     2701 ggttccggtg attttgatta tgaaaagatg gcaaacgcta ataagggggc tatgaccgaa
     2761 aatgccgatg aaaacgcgct acagtctgac gctaaaggca aacttgattc tgtcgctact
     2821 gattacggtg ctgctatcga tggtttcatt ggtgacgttt ccggccttgc taatggtaat
     2881 ggtgctactg gtgattttgc tggctctaat tcccaaatgg ctcaagtcgg tgacggtgat
     2941 aattcacctt taatgaataa tttccgtcaa tatttacctt ccctccctca atcggttgaa
     3001 tgtcgccctt ttgtctttgg cgctggtaaa ccttacgagt tcagtatcga ctgcgataag
     3061 atcaacctgt tccgcggtgt ctttgcgttt cttttatatg ttgccacctt tatgtatgta
     3121 ttttctacgt ttgctaacat actgcgtaat aaggagtctt aatgaaattt ggaaactttt
     3181 tgcttacata ccaacctccc caattttccc aaacagaggt aatgaaacgt ttggttaaat
     3241 taggtcgcat ctctgaggag tgtggttttg ataccgtatg gttactggag catcatttca
     3301 cggagtttgg tttgcttggt aacccttatg tcgctgctgc atatttactt ggcgcgacta
     3361 aaaaattgaa tgtaggaact gccgctattg ttcttcccac agcccatcca gtacgccaac
     3421 ttgaagatgt gaatttattg gatcaaatgt caaaaggacg atttcggttt ggtatttgcc
     3481 gagggcttta caacaaggac tttcgcgtat tcggcacaga tatgaataac agtcgcgcct
     3541 tagcggaatg ctggtacggg ctgataaaga atggcatgac agagggatat atggaagctg
     3601 ataatgaaca tatcaagttc cataaggtaa aagtaaaccc cgcggcgtat agcagaggtg
     3661 gcgcaccggt ttatgtggtg gctgaatcag cttcgacgac tgagtgggct gctcaatttg
     3721 gcctaccgat gatattaagt tggattataa atactaacga aaagaaagca caacttgagc
     3781 tttataatga agtggctcaa gaatatgggc acgatattca taatatcgac cattgcttat
     3841 catatataac atctgtagat catgactcaa ttaaagcgaa agagatttgc cggaaatttc
     3901 tggggcattg gtatgattct tatgtgaatg ctacgactat ttttgatgat tcagaccaaa
     3961 caagaggtta tgatttcaat aaagggcagt ggcgtgactt tgtattaaaa ggacataaag
     4021 atactaatcg ccgtattgat tacagttacg aaatcaatcc cgtgggaacg ccgcaggaat
     4081 gtattgacat aattcaaaaa gacattgatg ctacaggaat atcaaatatt tgttgtggat
     4141 ttgaagctaa tggaacagta gatgaaatta ttgcttccat gaagctcttc cagtctgatg
     4201 tcatgccatt tcttaaagaa aaacaacgtt cgctattata ttatggcggt ggcggtagcg
     4261 gcggtggcgg tagcggcggt ggcggtagcg gcggtggcgg tagcaaattt ggattgttct
     4321 tccttaactt catcaattca acaactgttc aagaacagag tatagttcgc atgcaggaaa
     4381 taacggagta tgttgataag ttgaattttg aacagatttt agtgtatgaa aatcattttt
     4441 cagataatgg tgttgtcggc gctcctctga ctgtttctgg ttttctgctc ggtttaacag
     4501 agaaaattaa aattggttca ttaaatcaca tcattacaac tcatcatcct gtccgcatag
     4561 cggaggaagc ttgcttattg gatcagttaa gtgaagggag atttatttta gggtttagtg
     4621 attgcgaaaa aaaagatgaa atgcattttt ttaatcgccc ggttgaatat caacagcaac
     4681 tatttgaaga gtgttatgaa atcattaacg atgctttaac aacaggctat tgtaatccag
     4741 ataacgattt ttatagcttc cctaaaatat ctgtaaatcc ccatgcttat acgccaggcg
     4801 gacctcggaa atatgtaaca gcaaccagtc atcatattgt tgagtgggcg gccaaaaaag
     4861 gtattcctct catctttaag tgggatgatt ctaatgatgt tagatatgaa tatgctgaaa
     4921 gatataaagc cgttgcggat aaatatgacg ttgacctatc agagatagac catcagttaa
     4981 tgatattagt taactataac gaagatagta ataaagctaa acaagagacg cgtgcattta
     5041 ttagtgatta tgttcttgaa atgcacccta atgaaaattt cgaaaataaa cttgaagaaa
     5101 taattgcaga aaacgctgtc ggaaattata cggagtgtat aactgcggct aagttggcaa
     5161 ttgaaaagtg tggtgcgaaa agtgtattgc tgtcctttga accaatgaat gatttgatga
     5221 gccaaaaaaa tgtaatcaat attgttgatg ataatattaa gaagtaccac acggaatata
     5281 cctaaacgga gccaatgtac gcaaaaaacc ccgcttcagc ggggtttttt cgccaaaaaa
     5341 aaccccgccc tgtcaggggc ggggtttttt tttagggccc tactgactgt tatgtgaggc
     5401 tgttgtccac gcccagctct gggctggagc cacccacaaa gtcctccaga tgcagctcgc
     5461 gagttaacag ggtttcctga atcgcacgca gcagtattgt gttactacgg agctgctcga
     5521 tctcgtcggt ggtggcgggc acgtggttcc gtatgaagtg cgcgcgcacc tcggtgcggt
     5581 cgtcgcgcgc ggcgccgctg ctgtccggcg cgcgcagcac cgcgtcgatg ttgcacgtca
     5641 ggctgaacac cgacgagaac gtgtccgcga tgaaagcacg gttggagtca acgacatcca
     5701 gcgtgttgtt gaacaagaag tagactctgt tgagatgagt gatcagatag atcttcacgt
     5761 cagtacgggc ttctgcacca cgggtgtctg tagccacgac atcgaactcg aacatgccga
     5821 ccatagtatc cagcgggttc atgttgaggg acaggacgcc gctgctcggg tgcagcgtga
     5881 aggcggactc acgcacgccc tccagcgagg ggtccaccac catcgtgtcc aggtccatcg
     5941 tgtacgttat gtctaggcct tcggtatgtg tcgcatgaac agtgagtagt tccctgttgg
     6001 atgtatccgt ctcgtagatg ccggcggtgt agatctcacg ctcgaatact ggccgcttgt
     6061 catcgtcgtc cttgtagtca acattcgaat cagaggtgcc attgagcagc cagtcaacac
     6121 ttacgccaag agctgacgca agttctggta aaaagcgtgg tcgcttagtt ttaccgtttt
     6181 cgagctgctc tatagactgc tgggtagtcc ccaccttttg agcaagttca gcctggttaa
     6241 gtccaagctg aattcttttg ctttttaccc tgctgctgat actagctcta accatgctaa
     6301 tcaatggata tttccctttg ggctcaacgt gcccaacaaa tctaacattc gaatcagagg
     6361 tgccattgag cagccagtca acacttacgc caagagctga cgcaagttct ggtaaaaagc
     6421 gtggtcgctt agttttaccg ttttcgagct gctctataga ctgctgggta gtccccacct
     6481 tttgagcaag ttcagcctgg ttaagtccaa gctgaattct tttgcttttt accctgctgc
     6541 tgatactcat ttttttttcc tccttactct agtaaaagtt aaacaaaatt atttgtagag
     6601 ggaaaccgtt gtggtctccc tgaatataga taccgagcct tatgcatgcc cgtaaagtta
     6661 tccagcaacc actcatagac ctagggcagc agatagggac gacgtggtgt tagctgtgct
     6721 ctagaatcct cagcgagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg
     6781 cttaatttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg
     6841 cttcgcaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga
     6901 caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt ttatttgatg
     6961 cctggcagtt ccctactctc gcatggggag accccacact accatcggcg ctacggcgtt
     7021 tcacttctga gttcggcatg gggtcaggtg ggaccaccgc gctactgccg ccaggcaaat
     7081 tctgttttat cagaccgctt ctgcgttctg atttaatctg tatcaggctg aaaatcttct
     7141 ctcatccgcc aaaacagcca tgcccatggc gtttaccaat gcttaatcag tgaggcacct
     7201 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7261 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7321 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7381 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7441 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7501 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7561 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7621 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7681 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7741 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7801 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7861 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7921 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7981 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8041 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8101 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8161 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     8221 aggcccttag gccaaatagg ccgt
//
