LOCUS       sequence-424405-        6534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses pIII.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6534)
  AUTHORS   Qi T, Wang Y, Yang Y, Gao S, Liu J, Huang Q, Tian Y, Tang J, Zheng 
            WV, Wang Y
  TITLE     Phage-assisted evolution of compact Cas9 variants targeting a simple
            NNG PAM.
  JOURNAL   Nat Chem Biol. 2023 Dec 5. doi: 10.1038/s41589-023-01481-5.
  PUBMED    38052959
REFERENCE   2  (bases 1 to 6534)
  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 Chem 
            Biol. 2023 Dec 5. doi: 10.1038/s41589-023-01481-5."
COMMENT     Sequence Label: pBT120a17-sg-del
FEATURES             Location/Qualifiers
     source          1..6534
                     /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"
     CDS             complement(join(3748..4539,4540..4608))
                     /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     4371..4390
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4609..4713)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4781..4799
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(4820..5770)
                     /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      5818..6040
                     /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 attaaaggcg cacttaatta acggcactgc
     3661 gaaaaaaccc cgccgaagcg gggttttttg cgaaatataa tgaccctctt gataacccaa
     3721 gagggcattt tttaatgccc atggcgttta ccaatgctta atcagtgagg cacctatctc
     3781 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3841 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgtg acccacgctc
     3901 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3961 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4021 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4081 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4141 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4201 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4261 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4321 agaatagtgt atgcggcgac cgagttgatc ttgcccggcg tcaatacggg ataataccgc
     4381 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4441 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4501 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4561 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4621 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4681 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     4741 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     4801 cttaggccaa ataggccgtt cagatccttc cgtatttagc cagtatgttc tctagtgtgg
     4861 ttcgttgttt ttgcgtgagc catgagaacg aaccattgag atcatgctta ctttgcatgt
     4921 cactcaaaaa ttttgcctca aaactggtga gctgaatttt tgcagttaaa gcatcgtgta
     4981 gtgtttttct tagtccgtta cgtaggtagg aatctgatgt aatggttgtt ggtattttgt
     5041 caccattcat ttttatctgg ttgttctcaa gttcggttac gagatccatt tgtctatcta
     5101 gttcaacttg gaaaatcaac gtatcagtcg ggcggcctcg cttatcaacc accaatttca
     5161 tattgctgta agtgtttaaa tctttactta ttggtttcaa aacccattgg ttaagccttt
     5221 taaactcatg gtagttattt tcaagcatta acatgaactt aaattcatca aggctaatct
     5281 ctatatttgc cttgtgagtt ttcttttgtg ttagttcttt taataaccac tcataaatcc
     5341 tcatagagta tttgttttca aaagacttaa catgttccag attatatttt atgaattttt
     5401 ttaactggaa aagataaggc aatatctctt cactaaaaac taattctaat ttttcgcttg
     5461 agaacttggc atagtttgtc cactggaaaa tcttaaagcc tttaaccaaa ggattcctga
     5521 tttccacagt tctcgtcatc agctctctgg ttgctttagc taatacacca taagcatttt
     5581 ccctactgat gttcatcatc tgagcgtatt ggttataagt gaacgatacc gtccgttctt
     5641 tccttgtagg gttttcaatc gtggggttga gtagtgccac acagcataaa attagcttgg
     5701 tttcatgctc cgttaagtca tagcgactaa tcgctagttc atttgctttg aaaacaacta
     5761 attcagacat acatctcaat tggtctaggt gattttaatc actataccaa ttgagatggg
     5821 ctagtcaatg ataattacta gtccttttcc tttgagttgt gggtatctgt aaattctgct
     5881 agacctttgc tggaaaactt gtaaattctg ctagaccctc tgtaaattcc gctagacctt
     5941 tgtgtgtttt ttttgtttat attcaagtgg ttataattta tagaataaag aaagaataaa
     6001 aaaagataaa aagaatagat cccagccctg tgtataactc actactttag tcagttccgc
     6061 agtattacaa aaggatgtcg caaacgctgt ttgctcctct acaaaacaga ccttaaaacc
     6121 ctaaaggctt aagtagcacc ctcgcaagct cgggcaaatc gctgaatatt ccttttgtct
     6181 ccgaccatca ggcacctgag tcgctgtctt tttcgtgaca ttcagttcgc tgcgctcacg
     6241 gctctggcag tgaatggggg taaatggcac tacaggcgcc ttttatggat tcatgcaagg
     6301 aaactaccca taatacaaga aaagcccgtc acgggcttct cagggcgttt tatggcgggt
     6361 ctgctatgtg gtgctatctg actttttgct gttcagcagt tcctgccctc tgattttcca
     6421 gtctgaccac ttcggattat cccgtgacag gtcattcaga ctggctaatg cacccagtaa
     6481 ggcagcggta tcatcaacag gcttacccgt cttactgtca agaggacatc cggt
//
