LOCUS       sequence-262576-        8706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ABEmax and nSauriCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8706)
  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 8706)
  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: SauriABEmax
FEATURES             Location/Qualifiers
     source          1..8706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     286..306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     377..396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             445..465
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4876..4896
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4906..4962
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(4963..4965,4966..4968,4969..5679)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4963..4965,4966..4968,4969..5679)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5008..5029)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5269..5288)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5616..5637
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5692..5712
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5721..5738
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5761..5778)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5767..5991
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6062..6078)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6062..6078)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6075..6097)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6086..6102
                     /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(6110..6116,6117..6134,6135..6140))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6155..6176
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6293..6310)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6464..7052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6544..6563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7223..8014,8015..8083))
                     /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     7846..7865
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8084..8188)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8267..8286)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(8458..8706,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat gaaacggaca
      421 gccgacggaa gcgagttcga gtcaccaaag aagaagcgga aagtctctga agtcgagttt
      481 agccacgagt attggatgag gcacgcactg accctggcaa agcgagcatg ggatgaaaga
      541 gaagtccccg tgggcgccgt gctggtgcac aacaatagag tgatcggaga gggatggaac
      601 aggccaatcg gccgccacga ccctaccgca cacgcagaga tcatggcact gaggcaggga
      661 ggcctggtca tgcagaatta ccgcctgatc gatgccaccc tgtatgtgac actggagcca
      721 tgcgtgatgt gcgcaggagc aatgatccac agcaggatcg gaagagtggt gttcggagca
      781 cgggacgcca agaccggcgc agcaggctcc ctgatggatg tgctgcacca ccccggcatg
      841 aaccaccggg tggagatcac agagggaatc ctggcagacg agtgcgccgc cctgctgagc
      901 gatttcttta gaatgcggag acaggagatc aaggcccaga agaaggcaca gagctccacc
      961 gactctggag gatctagcgg aggatcctct ggaagcgaga caccaggcac aagcgagtcc
     1021 gccacaccag agagctccgg cggctcctcc ggaggatcct ctgaggtgga gttttcccac
     1081 gagtactgga tgagacatgc cctgaccctg gccaagaggg cacgcgatga gagggaggtg
     1141 cctgtgggag ccgtgctggt gctgaacaat agagtgatcg gcgagggctg gaacagagcc
     1201 atcggcctgc acgacccaac agcccatgcc gaaattatgg ccctgagaca gggcggcctg
     1261 gtcatgcaga actacagact gattgacgcc accctgtacg tgacattcga gccttgcgtg
     1321 atgtgcgccg gcgccatgat ccactctagg atcggccgcg tggtgtttgg cgtgaggaac
     1381 gcaaaaaccg gcgccgcagg ctccctgatg gacgtgctgc actaccccgg catgaatcac
     1441 cgcgtcgaaa ttaccgaggg aatcctggca gatgaatgtg ccgccctgct gtgctatttc
     1501 tttcggatgc ctagacaggt gttcaatgct cagaagaagg cccagagctc caccgactcc
     1561 ggaggatcta gcggaggctc ctctggctct gagacacctg gcacaagcga gagcgcaaca
     1621 cctgaaagca gcgggggcag cagcgggggg tcacaggaga accagcagaa gcagaactac
     1681 atcctgggcc tggccatcgg aatcaccagc gtcggctacg gactgatcga tagcaagaca
     1741 agagaagtga tcgacgccgg cgttagactc tttccagaag ctgatagcga gaacaactcc
     1801 aaccgcagaa gcaagcgggg cgccagacgg ttaaaacgga gaagaatcca ccggctgaac
     1861 cgggtcaaag acctgctcgc tgattaccag atgatcgatc ttaacaatgt tcctaagagc
     1921 accgacccct acaccatcag agtgaagggc ctccgggagc ctctgacaaa agaagaattc
     1981 gccatcgccc tcctgcatat cgctaagaga agaggcctgc acaacatcag tgtgtccatg
     2041 ggcgacgaag agcaggacaa tgaactgagc accaagcagc agctgcaaaa gaatgcccag
     2101 caactgcagg acaagtatgt gtgcgaactg cagttagaac ggctgaccaa catcaacaag
     2161 gtcagaggcg agaagaacag atttaagaca gaggactttg tgaaagaagt gaaacagctg
     2221 tgcgaaaccc agagacagta ccacaacatc gacgaccaat tcatccagca gtacatcgac
     2281 ctggtgtcta caagacggga gtacttcgag ggccccggca acggctctcc atacggctgg
     2341 gacggcgacc tgctgaagtg gtacgagaag ctgatgggca gatgcaccta tttccccgaa
     2401 gaactgaggt ccgtgaagta cgcctacagc gccgacctct tcaacgccct gaacgacctg
     2461 aacaacctcg ttgtgaccag ggatgacaat ccaaagcttg agtactacga gaagtaccac
     2521 attattgaga acgtgttcaa gcaaaagaag aatcccacac tcaaacaaat cgccaaagag
     2581 atcggcgtgc aagattacga catccggggc tatagaatca caaagagcgg caaacctcag
     2641 ttcacctctt ttaagctgta tcacgacctg aagaacatct tcgagcaggc caaatacctg
     2701 gaagatgtgg aaatgctgga cgagatcgcc aagatcctga ccatctacca ggatgagatt
     2761 agcatcaaga aagccctgga ccagctgccc gaactgctga cagagagcga gaaatctcag
     2821 atcgcacagc tcaccggcta tacaggcacc cacagactga gcctgaagtg catccacatt
     2881 gtgatcgacg agctgtggga gagccccgag aaccagatgg aaatctttac cagactgaat
     2941 ctgaaaccta agaaggtgga aatgagcgag atcgacagca tacccaccac cctggtcgac
     3001 gagttcatcc tctcacctgt ggtgaagcgg gccttcatcc agagcatcaa ggtaatcaac
     3061 gcagtgatca atcggttcgg cctgccagag gacatcatca tcgagctggc cagagaaaag
     3121 aatagcaagg atcggagaaa gttcattaac aagctgcaga aacaaaatga ggccacaaga
     3181 aagaaaatcg aacagctgct ggccaagtac ggcaacacca atgccaagta catgatcgag
     3241 aagatcaagc tgcacgacat gcaggagggc aagtgcctgt acagcctgga ggctattcct
     3301 ctggaagacc tgctgagcaa cccgacacac tacgaagttg accacattat ccccagatct
     3361 gtgagctttg acaacagcct gaacaacaaa gtgctggtga aacaaagcga aaacagcaag
     3421 aagggcaatc gcacccctta ccagtacctg agcagcaacg agtctaagat tagctacaac
     3481 cagtttaagc agcacatcct gaacctgagc aaggccaagg acagaatcag caagaaaaaa
     3541 agagatatgc tgctggaaga gagagatatc aacaagttcg aagtgcagaa ggaattcatt
     3601 aaccggaacc tggtggatac acggtacgcc accagagaac tgtctaacct gctgaagacc
     3661 tacttcagca cccatgacta cgccgtgaag gtgaagacca tcaacggcgg cttcactaac
     3721 cacctgagga aggtgtggga tttcaagaag cacagaaacc acggctacaa gcaccacgcc
     3781 gaagatgccc tggtgatcgc caacgccgac ttcctgttta agacacataa ggccctgcgg
     3841 agaaccgata agatcctgga acaacctggc ctggaagtga atgatacaac cgtgaaagtg
     3901 gacaccgagg aaaaatacca ggagctgttc gagacaccta agcaagtgaa gaacatcaag
     3961 cagttccggg acttcaagta cagccaccga gtggacaaga agcctaaccg gcagcttatc
     4021 aacgacacac tgtactccac cagagagatt gatggcgaaa cctacgtggt gcagaccctt
     4081 aaggatctgt acgccaagga caacgagaaa gtgaagaagc tgttcaccga aagacctcag
     4141 aagatcctga tgtaccagca cgaccctaag accttcgaga aactgatgac aatcctgaac
     4201 cagtacgctg aggccaagaa ccctctggct gcttattacg aggacaaagg cgagtacgtg
     4261 accaagtacg ccaagaaagg caatggacct gccatccaca agatcaagta tatcgataag
     4321 aagcttggat cttacctgga tgttagcaac aagtatcctg agacacagaa caagcttgtg
     4381 aagctgtccc tgaagagctt tagattcgac atctacaagt gtgaacaggg ctacaagatg
     4441 gtgtccatcg gatacctgga cgtgctgaag aaagataact actactacat ccctaaggac
     4501 aagtacgagg ccgagaagca gaaaaagaag atcaaggaat ctgatctttt tgtgggcagc
     4561 ttctactaca acgacctcat catgtacgag gatgaactgt tcagagtgat aggagtgaac
     4621 agcgacatca acaatctggt tgagctaaac atggtcgaca ttacctacaa ggacttctgc
     4681 gaggtgaaca acgtgacagg cgagaaaaga atcaaaaaga ctatcggcaa gcgcgtggtc
     4741 ctgatcgaga agtacaccac agatattcta ggcaacctgt acaagactcc cctgcctaag
     4801 aagccccagc ttatcttcaa gcggggagaa ctgtctggcg gctcaaaaag aaccgccgac
     4861 ggcagcgaat tcgagcccaa gaagaagagg aaagtcggaa gcggagctac taacttcagc
     4921 ctgctgaagc aggctggaga cgtggaggag aaccctggac ctatggtgag caagggcgag
     4981 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     5041 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     5101 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     5161 tatggagtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     5221 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     5281 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     5341 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     5401 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     5461 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     5521 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     5581 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     5641 gccgccggga tcactctcgg catggacgag ctgtacaagt ctggtggttc tcccaagaag
     5701 aagaggaaag tctaaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag
     5761 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     5821 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     5881 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     5941 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     6001 cggaaagaac cagctggggc tcgataccgt cgacctctag ctagagcttg gcgtaatcat
     6061 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6121 ccggaagcat aaagtgtaaa gcctagggtg cctaatgagt gagctaactc acattaattg
     6181 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6241 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6301 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6361 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6421 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6481 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6541 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6601 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6661 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6721 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6781 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6841 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6901 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6961 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7021 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7081 ctgacactca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7141 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7201 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7261 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7321 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7381 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7441 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7501 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7561 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7621 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7681 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7741 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7801 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7861 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7921 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7981 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8041 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8101 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8161 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     8221 acggatcggg agatcgatct cccgatcccc tagggtcgac tctcagtaca atctgctctg
     8281 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     8341 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     8401 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
     8461 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     8521 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     8581 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     8641 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     8701 tgtatc
//
