LOCUS       sequence-292628-        8049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AncBE4max and nSauriCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8049)
  AUTHORS   Hu Z, Wang S, Zhang C, Gao N, Li M, Wang D, Wang D, Liu D, Liu H, 
            Ong SG, Wang H, Wang Y
  TITLE     A compact Cas9 ortholog from Staphylococcus Auricularis (SauriCas9) 
            expands the DNA targeting scope.
  JOURNAL   PLoS Biol. 2020 Mar 30;18(3):e3000686. doi: 
            10.1371/journal.pbio.3000686. eCollection 2020 Mar.
  PUBMED    32226015
REFERENCE   2  (bases 1 to 8049)
  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: "PLoS 
            Biol."; date: "2020-03-30"; volume: "18(3):e3000686. doi"; pages: " 
            10.1371/journal.pbio.3000686. eCollection 2020 Mar"
COMMENT     Sequence Label: SauriBE4max
FEATURES             Location/Qualifiers
     source          1..8049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             694..714
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4714..4962
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             4993..5241
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5284..5304
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5313..5330
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5353..5370)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5359..5583
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5654..5670)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5654..5670)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5667..5689)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5678..5694
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5702..5708,5709..5726,5727..5732))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5747..5768
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5885..5902)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6056..6644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6136..6155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6815..7606,7607..7675))
                     /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     7438..7457
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7676..7780)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7859..7878)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      601 agatccgcta gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg
      661 aaacggacag ccgacggaag cgagttcgag tcaccaaaga agaagcggaa agtcagcagt
      721 gaaaccggac cagtggcagt ggacccaacc ctgaggagac ggattgagcc ccatgaattt
      781 gaagtgttct ttgacccaag ggagctgagg aaggagacat gcctgctgta cgagatcaag
      841 tggggcacaa gccacaagat ctggcgccac agctccaaga acaccacaaa gcacgtggaa
      901 gtgaatttca tcgagaagtt tacctccgag cggcacttct gcccctctac cagctgttcc
      961 atcacatggt ttctgtcttg gagcccttgc ggcgagtgtt ccaaggccat caccgagttc
     1021 ctgtctcagc accctaacgt gaccctggtc atctacgtgg cccggctgta tcaccacatg
     1081 gaccagcaga acaggcaggg cctgcgcgat ctggtgaatt ctggcgtgac catccagatc
     1141 atgacagccc cagagtacga ctattgctgg cggaacttcg tgaattatcc acctggcaag
     1201 gaggcacact ggccaagata cccacccctg tggatgaagc tgtatgcact ggagctgcac
     1261 gcaggaatcc tgggcctgcc tccatgtctg aatatcctgc ggagaaagca gccccagctg
     1321 acatttttca ccattgctct gcagtcttgt cactatcagc ggctgcctcc tcatattctg
     1381 tgggctacag gcctgaagtc tggaggatct agcggaggat cctctggcag cgagacacca
     1441 ggaacaagcg agtcagcaac accagagagc agtggcggca gcagcggcgg cagccaggag
     1501 aaccagcaga agcagaacta catcctgggc ctggccatcg gaatcaccag cgtcggctac
     1561 ggactgatcg atagcaagac aagagaagtg atcgacgccg gcgttagact ctttccagaa
     1621 gctgatagcg agaacaactc caaccgcaga agcaagcggg gcgccagacg gttaaaacgg
     1681 agaagaatcc accggctgaa ccgggtcaaa gacctgctcg ctgattacca gatgatcgat
     1741 cttaacaatg ttcctaagag caccgacccc tacaccatca gagtgaaggg cctccgggag
     1801 cctctgacaa aagaagaatt cgccatcgcc ctcctgcata tcgctaagag aagaggcctg
     1861 cacaacatca gtgtgtccat gggcgacgaa gagcaggaca atgaactgag caccaagcag
     1921 cagctgcaaa agaatgccca gcaactgcag gacaagtatg tgtgcgaact gcagttagaa
     1981 cggctgacca acatcaacaa ggtcagaggc gagaagaaca gatttaagac agaggacttt
     2041 gtgaaagaag tgaaacagct gtgcgaaacc cagagacagt accacaacat cgacgaccaa
     2101 ttcatccagc agtacatcga cctggtgtct acaagacggg agtacttcga gggccccggc
     2161 aacggctctc catacggctg ggacggcgac ctgctgaagt ggtacgagaa gctgatgggc
     2221 agatgcacct atttccccga agaactgagg tccgtgaagt acgcctacag cgccgacctc
     2281 ttcaacgccc tgaacgacct gaacaacctc gttgtgacca gggatgacaa tccaaagctt
     2341 gagtactacg agaagtacca cattattgag aacgtgttca agcaaaagaa gaatcccaca
     2401 ctcaaacaaa tcgccaaaga gatcggcgtg caagattacg acatccgggg ctatagaatc
     2461 acaaagagcg gcaaacctca gttcacctct tttaagctgt atcacgacct gaagaacatc
     2521 ttcgagcagg ccaaatacct ggaagatgtg gaaatgctgg acgagatcgc caagatcctg
     2581 accatctacc aggatgagat tagcatcaag aaagccctgg accagctgcc cgaactgctg
     2641 acagagagcg agaaatctca gatcgcacag ctcaccggct atacaggcac ccacagactg
     2701 agcctgaagt gcatccacat tgtgatcgac gagctgtggg agagccccga gaaccagatg
     2761 gaaatcttta ccagactgaa tctgaaacct aagaaggtgg aaatgagcga gatcgacagc
     2821 atacccacca ccctggtcga cgagttcatc ctctcacctg tggtgaagcg ggccttcatc
     2881 cagagcatca aggtaatcaa cgcagtgatc aatcggttcg gcctgccaga ggacatcatc
     2941 atcgagctgg ccagagaaaa gaatagcaag gatcggagaa agttcattaa caagctgcag
     3001 aaacaaaatg aggccacaag aaagaaaatc gaacagctgc tggccaagta cggcaacacc
     3061 aatgccaagt acatgatcga gaagatcaag ctgcacgaca tgcaggaggg caagtgcctg
     3121 tacagcctgg aggctattcc tctggaagac ctgctgagca acccgacaca ctacgaagtt
     3181 gaccacatta tccccagatc tgtgagcttt gacaacagcc tgaacaacaa agtgctggtg
     3241 aaacaaagcg aaaacagcaa gaagggcaat cgcacccctt accagtacct gagcagcaac
     3301 gagtctaaga ttagctacaa ccagtttaag cagcacatcc tgaacctgag caaggccaag
     3361 gacagaatca gcaagaaaaa aagagatatg ctgctggaag agagagatat caacaagttc
     3421 gaagtgcaga aggaattcat taaccggaac ctggtggata cacggtacgc caccagagaa
     3481 ctgtctaacc tgctgaagac ctacttcagc acccatgact acgccgtgaa ggtgaagacc
     3541 atcaacggcg gcttcactaa ccacctgagg aaggtgtggg atttcaagaa gcacagaaac
     3601 cacggctaca agcaccacgc cgaagatgcc ctggtgatcg ccaacgccga cttcctgttt
     3661 aagacacata aggccctgcg gagaaccgat aagatcctgg aacaacctgg cctggaagtg
     3721 aatgatacaa ccgtgaaagt ggacaccgag gaaaaatacc aggagctgtt cgagacacct
     3781 aagcaagtga agaacatcaa gcagttccgg gacttcaagt acagccaccg agtggacaag
     3841 aagcctaacc ggcagcttat caacgacaca ctgtactcca ccagagagat tgatggcgaa
     3901 acctacgtgg tgcagaccct taaggatctg tacgccaagg acaacgagaa agtgaagaag
     3961 ctgttcaccg aaagacctca gaagatcctg atgtaccagc acgaccctaa gaccttcgag
     4021 aaactgatga caatcctgaa ccagtacgct gaggccaaga accctctggc tgcttattac
     4081 gaggacaaag gcgagtacgt gaccaagtac gccaagaaag gcaatggacc tgccatccac
     4141 aagatcaagt atatcgataa gaagcttgga tcttacctgg atgttagcaa caagtatcct
     4201 gagacacaga acaagcttgt gaagctgtcc ctgaagagct ttagattcga catctacaag
     4261 tgtgaacagg gctacaagat ggtgtccatc ggatacctgg acgtgctgaa gaaagataac
     4321 tactactaca tccctaagga caagtacgag gccgagaagc agaaaaagaa gatcaaggaa
     4381 tctgatcttt ttgtgggcag cttctactac aacgacctca tcatgtacga ggatgaactg
     4441 ttcagagtga taggagtgaa cagcgacatc aacaatctgg ttgagctaaa catggtcgac
     4501 attacctaca aggacttcta cgaggtgaac aacgtgacag gcgagaaaag aatcaaaaag
     4561 actatcggca agcgcgtggt cctgatcgag aagtacacca cagatattct aggcaacctg
     4621 tacaagactc ccctgcctaa gaagccccag cttatcttca agcgggaaga actgagcggc
     4681 gggagcggcg ggagcggcgg gagcgggggg agcactaatc tgagcgacat cattgagaag
     4741 gagactggga aacagctggt cattcaggag tccatcctga tgctgcctga ggaggtggag
     4801 gaagtgatcg gcaacaagcc agagtctgac atcctggtgc acaccgccta cgacgagtcc
     4861 acagatgaga atgtgatgct gctgacctct gacgcccccg agtataagcc ttgggccctg
     4921 gtcatccagg attctaacgg cgagaataag atcaagatgc tgagcggagg atccggagga
     4981 tctggaggca gcaccaacct gtctgacatc atcgagaagg agacaggcaa gcagctggtc
     5041 atccaggaga gcatcctgat gctgcccgaa gaagtcgaag aagtgatcgg aaacaagcct
     5101 gagagcgata tcctggtcca taccgcctac gacgagagta ccgacgaaaa tgtgatgctg
     5161 ctgacatccg acgccccaga gtataagccc tgggctctgg tcatccagga ttccaacgga
     5221 gagaacaaaa tcaaaatgct gtctggcggc tcaaaaagaa ccgccgacgg cagcgaattc
     5281 gagcccaaga agaagaggaa agtctaaccg gtcatcatca ccatcaccat tgagtttaaa
     5341 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     5401 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     5461 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     5521 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     5581 tggcttctga ggcggaaaga accagctggg gctcgatacc gtcgacctct agctagagct
     5641 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5701 acaacatacg agccggaagc ataaagtgta aagcctagga tgcctaatga gtgagctaac
     5761 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5821 tgcattaatg aatcggccaa cgcgcgggaa gaggcggttt gcgtattggg cgctcttccg
     5881 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5941 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6001 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6061 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6121 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6181 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6241 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6301 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6361 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6421 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6481 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6541 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6601 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6661 tttctacggg gtctgacact cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6721 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6781 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6841 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6901 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6961 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7021 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7081 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7141 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7201 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7261 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7321 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7381 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7441 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7501 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7561 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7621 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7681 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7741 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7801 cacctgacgt cgacggatcg ggagatcgat ctcccgatcc cctagggtcg actctcagta
     7861 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     7921 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     7981 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
     8041 atacgcgtt
//
