LOCUS       sequence-238890-        6808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BE PACE selection, express gIII, luxAB and degron-GTCCA guide RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6808)
  AUTHORS   Thuronyi BW, Koblan LW, Levy JM, Yeh WH, Zheng C, Newby GA, Wilson 
            C, Bhaumik M, Shubina-Oleinik O, Holt JR, Liu DR
  TITLE     Continuous evolution of base editors with expanded target 
            compatibility and improved activity.
  JOURNAL   Nat Biotechnol. 2019 Jul 22. pii: 10.1038/s41587-019-0193-0. doi: 
            10.1038/s41587-019-0193-0.
  PUBMED    31332326
REFERENCE   2  (bases 1 to 6808)
  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"; journalName: "Nat 
            Biotechnol. 2019 Jul 22. pii: 10.1038/s41587-019-0193-0. doi: 
            10.1038/s41587-019-0193-0."
COMMENT     Sequence Label: pBT120b
FEATURES             Location/Qualifiers
     source          1..6808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     70..89
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        70..88
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1399..2478
                     /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             2542..3522
                     /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"
     misc_RNA        complement(3642..3717)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(join(3744..3750,3751..3768,3769..3774))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3789..3810
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(join(4022..4813,4814..4882))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQDQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4645..4664
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4883..4987)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5055..5073
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(5094..6044)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFKIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      6092..6314
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttgctga ggagacttag
       61 ggaccctact aatacgactc actataggga gaaagacctg caggtgcagt aaaggaaaaa
      121 aaaaatgaaa aaattattat tcgcaattcc tttagttgtt cctttctatt ctcactccgc
      181 tgaaactgtt gaaagttgtt tagcaaaacc ccatacagaa aattcattta ctaacgtctg
      241 gaaagacgac aaaactttag atcgttacgc taactatgag ggctgtctgt ggaatgctac
      301 aggcgttgta gtttgtactg gtgacgaaac tcagtgttac ggtacatggg ttcctattgg
      361 gcttgctatc cctgaaaatg agggtggtgg ctctgagggt ggtggttctg agggtggcgg
      421 ttctgagggt ggcggtacta aacctcctga gtacggtgat acacctattc cgggctatac
      481 ttatatcaac cctctcgacg gcacttatcc gcctggtact gagcaaaacc ccgctaatcc
      541 taatccttct cttgaggagt ctcagcctct taatactttc atgtttcaga ataataggtt
      601 ccgaaatagg cagggggcat taactgttta tacgggcact gttactcaag gcactgaccc
      661 cgttaaaact tattaccagt acactcctgt atcatcaaaa gccatgtatg acgcttactg
      721 gaacggtaaa ttcagagact gcgctttcca ttctggcttt aatgaggatc cattcgtttg
      781 tgaatatcaa ggccaatcgt ctgacctgcc tcaacctcct gtcaatgctg gcggcggctc
      841 tggtggtggt tctggtggcg gctctgaggg tggtggctct gagggtggcg gttctgaggg
      901 tggcggctct gagggaggcg gttccggtgg tggctctggt tccggtgatt ttgattatga
      961 aaagatggca aacgctaata agggggctat gaccgaaaat gccgatgaaa acgcgctaca
     1021 gtctgacgct aaaggcaaac ttgattctgt cgctactgat tacggtgctg ctatcgatgg
     1081 tttcattggt gacgtttccg gccttgctaa tggtaatggt gctactggtg attttgctgg
     1141 ctctaattcc caaatggctc aagtcggtga cggtgataat tcacctttaa tgaataattt
     1201 ccgtcaatat ttacctagtc ttccgcagag tgtagagtgc cgaccattcg ttttcggtgc
     1261 gggcaagccg tacgagttca gcatcgactg cgataagatc aatttgtttc gaggagtttt
     1321 cgcattccta ctctacgtgg caacgttcat gtacgtgttc tcaacattcg ccaatatttt
     1381 aagaaacaaa gagtcttaat gaaatttgga aactttttgc ttacatacca acctccccaa
     1441 ttttcccaaa cagaggtaat gaaacgtttg gttaaattag gtcgcatctc tgaggagtgt
     1501 ggttttgata ccgtatggtt actggagcat catttcacgg agtttggttt gcttggtaac
     1561 ccttatgtcg ctgctgcata tttacttggc gcgactaaaa aattgaatgt aggaactgcc
     1621 gctattgttc ttcccacagc ccatccagta cgccaacttg aagatgtgaa tttattggat
     1681 caaatgtcaa aaggacgatt tcggtttggt atttgccgag ggctttacaa caaggacttt
     1741 cgcgtattcg gcacagatat gaataacagt cgcgccttag cggaatgctg gtacgggctg
     1801 ataaagaatg gcatgacaga gggatatatg gaagctgata atgaacatat caagttccat
     1861 aaggtaaaag taaaccccgc ggcgtatagc agaggtggcg caccggttta tgtggtggct
     1921 gaatcagctt cgacgactga gtgggctgct caatttggcc taccgatgat attaagttgg
     1981 attataaata ctaacgaaaa gaaagcacaa cttgagcttt ataatgaagt ggctcaagaa
     2041 tatgggcacg atattcataa tatcgaccat tgcttatcat atataacatc tgtagatcat
     2101 gactcaatta aagcgaaaga gatttgccgg aaatttctgg ggcattggta tgattcttat
     2161 gtgaatgcta cgactatttt tgatgattca gaccaaacaa gaggttatga tttcaataaa
     2221 gggcagtggc gtgactttgt attaaaagga cataaagata ctaatcgccg tattgattac
     2281 agttacgaaa tcaatcccgt gggaacgccg caggaatgta ttgacataat tcaaaaagac
     2341 attgatgcta caggaatatc aaatatttgt tgtggatttg aagctaatgg aacagtagac
     2401 gaaattattg cttccatgaa gctgttccag tctgatgtca tgccatttct taaagaaaaa
     2461 caacgttcgc tattatatta tggcggtggc ggtagcggcg gtggcggtag cggcggtggc
     2521 ggtagcggcg gtggcggtag caaatttgga ttgttcttcc ttaacttcat caattcaaca
     2581 actgttcaag aacagagtat agttcgcatg caggaaataa cggagtatgt tgataagttg
     2641 aattttgaac agattttagt gtatgaaaat catttttcag ataatggtgt tgtcggcgct
     2701 cctctgactg tttctggttt tctgctcggt ttaacagaga aaattaaaat tggttcatta
     2761 aatcacatca ttacaactca tcatcctgtc cgcatagcgg aggaagcttg cttattggat
     2821 cagttaagtg aagggagatt tattttaggg tttagtgatt gcgaaaaaaa agatgaaatg
     2881 cattttttta atcgcccggt tgaatatcaa cagcaactat ttgaagagtg ttatgaaatc
     2941 attaacgatg ctttaacaac aggctattgt aatccagata acgattttta tagcttccct
     3001 aaaatatctg taaatcccca tgcttatacg ccaggcggac ctcggaaata tgtaacagca
     3061 accagtcatc atattgttga gtgggcggcc aaaaaaggta ttcctctcat ctttaagtgg
     3121 gatgattcta atgatgttag atatgaatat gctgaaagat ataaagccgt tgcggataaa
     3181 tatgacgttg acctatcaga gatagaccat cagttaatga tattagttaa ctataacgaa
     3241 gatagtaata aagctaaaca agagacgcgt gcatttatta gtgattatgt tcttgaaatg
     3301 caccctaatg aaaatttcga aaataaactt gaagaaataa ttgcagaaaa cgctgtcgga
     3361 aattatacgg agtgtataac tgcggctaag ttggcaattg aaaagtgtgg tgcgaaaagt
     3421 gtattgctgt cctttgaacc aatgaatgat ttgatgagcc aaaaaaatgt aatcaatatt
     3481 gttgatgata atattaagaa gtaccacacg gaatatacct aaacggagcc aatgtacgca
     3541 aaaaccccgc ttcagcgggg ttttttcgcc aaaaaaaacc ccgccctgtc aggggcgggg
     3601 ttttttttta tgttgaaaat ctccttctag attaaaaaaa agcaccgact cggtgccact
     3661 ttttcaagtt gataacggac tagccttatt ttaacttgct atttctagct ctaaaacact
     3721 tcgcgttcgc ctggacctcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     3781 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3841 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3901 ttgcgtattg ggcgcactta attaacggca ctgcgaaaaa accccgccga agcggggttt
     3961 tttgcgaaat ataatgaccc tcttgataac ccaagagggc attttttaat gcccatggcg
     4021 tttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4081 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4141 ccagtgctgc aatgataccg cgtgacccac gctcaccggc tccagattta tcagcaataa
     4201 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4261 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4321 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4381 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4441 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4501 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4561 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4621 gatcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4681 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4741 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4801 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4861 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4921 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4981 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     5041 cattaaccta taaaaatagg cgtatcacga ggcccttagg ccaaataggc cgttcagatc
     5101 cttccgtatt tagccagtat gttctctagt gtggttcgtt gtttttgcgt gagccatgag
     5161 aacgaaccat tgagatcatg cttactttgc atgtcactca aaaattttgc ctcaaaactg
     5221 gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt ttcttagtcc gttacgtagg
     5281 taggaatctg atgtaatggt tgttggtatt ttgtcaccat tcatttttat ctggttgttc
     5341 tcaagttcgg ttacgagatc catttgtcta tctagttcaa cttggaaaat caacgtatca
     5401 gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc tgtaagtgtt taaatcttta
     5461 cttattggtt tcaaaaccca ttggttaagc cttttaaact catggtagtt attttcaagc
     5521 attaacatga acttaaattc atcaaggcta atctctatat ttgccttgtg agttttcttt
     5581 tgtgttagtt cttttaataa ccactcataa atcctcatag agtatttgtt ttcaaaagac
     5641 ttaacatgtt ccagattata ttttatgaat ttttttaact ggaaaagata aggcaatatc
     5701 tcttcactaa aaactaattc taatttttcg cttgagaact tggcatagtt tgtccactgg
     5761 aaaatcttaa agcctttaac caaaggattc ctgatttcca cagttctcgt catcagctct
     5821 ctggttgctt tagctaatac accataagca ttttccctac tgatgttcat catctgagcg
     5881 tattggttat aagtgaacga taccgtccgt tctttccttg tagggttttc aatcgtgggg
     5941 ttgagtagtg ccacacagca taaaattagc ttggtttcat gctccgttaa gtcatagcga
     6001 ctaatcgcta gttcatttgc tttgaaaaca actaattcag acatacatct caattggtct
     6061 aggtgatttt aatcactata ccaattgaga tgggctagtc aatgataatt actagtcctt
     6121 ttcctttgag ttgtgggtat ctgtaaattc tgctagacct ttgctggaaa acttgtaaat
     6181 tctgctagac cctctgtaaa ttccgctaga cctttgtgtg ttttttttgt ttatattcaa
     6241 gtggttataa tttatagaat aaagaaagaa taaaaaaaga taaaaagaat agatcccagc
     6301 cctgtgtata actcactact ttagtcagtt ccgcagtatt acaaaaggat gtcgcaaacg
     6361 ctgtttgctc ctctacaaaa cagaccttaa aaccctaaag gcttaagtag caccctcgca
     6421 agctcgggca aatcgctgaa tattcctttt gtctccgacc atcaggcacc tgagtcgctg
     6481 tctttttcgt gacattcagt tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg
     6541 gcactacagg cgccttttat ggattcatgc aaggaaacta cccataatac aagaaaagcc
     6601 cgtcacgggc ttctcagggc gttttatggc gggtctgcta tgtggtgcta tctgactttt
     6661 tgctgttcag cagttcctgc cctctgattt tccagtctga ccacttcgga ttatcccgtg
     6721 acaggtcatt cagactggct aatgcaccca gtaaggcagc ggtatcatca acaggcttac
     6781 ccgtcttact gtcaagagga catccggt
//
