LOCUS       sequence-287974-        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"
     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             5125..5145
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5155..5211
                     /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(5212..5214,5215..5217,5218..5928)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5212..5214,5215..5217,5218..5928)
                     /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(5257..5278)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5518..5537)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5865..5886
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5941..5961
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5970..5987
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6010..6027)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6016..6240
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6311..6327)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6311..6327)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6324..6346)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6335..6351
                     /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(6359..6365,6366..6383,6384..6389))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6404..6425
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6542..6559)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6713..7301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6793..6812)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7472..8263,8264..8332))
                     /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     8095..8114
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8333..8437)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8516..8535)
                     /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 agtctctgaa
      721 gtcgagttta gccacgagta ttggatgagg cacgcactga ccctggcaaa gcgagcatgg
      781 gatgaaagag aagtccccgt gggcgccgtg ctggtgcaca acaatagagt gatcggagag
      841 ggatggaaca ggccaatcgg ccgccacgac cctaccgcac acgcagagat catggcactg
      901 aggcagggag gcctggtcat gcagaattac cgcctgatcg atgccaccct gtatgtgaca
      961 ctggagccat gcgtgatgtg cgcaggagca atgatccaca gcaggatcgg aagagtggtg
     1021 ttcggagcac gggacgccaa gaccggcgca gcaggctccc tgatggatgt gctgcaccac
     1081 cccggcatga accaccgggt ggagatcaca gagggaatcc tggcagacga gtgcgccgcc
     1141 ctgctgagcg atttctttag aatgcggaga caggagatca aggcccagaa gaaggcacag
     1201 agctccaccg actctggagg atctagcgga ggatcctctg gaagcgagac accaggcaca
     1261 agcgagtccg ccacaccaga gagctccggc ggctcctccg gaggatcctc tgaggtggag
     1321 ttttcccacg agtactggat gagacatgcc ctgaccctgg ccaagagggc acgcgatgag
     1381 agggaggtgc ctgtgggagc cgtgctggtg ctgaacaata gagtgatcgg cgagggctgg
     1441 aacagagcca tcggcctgca cgacccaaca gcccatgccg aaattatggc cctgagacag
     1501 ggcggcctgg tcatgcagaa ctacagactg attgacgcca ccctgtacgt gacattcgag
     1561 ccttgcgtga tgtgcgccgg cgccatgatc cactctagga tcggccgcgt ggtgtttggc
     1621 gtgaggaacg caaaaaccgg cgccgcaggc tccctgatgg acgtgctgca ctaccccggc
     1681 atgaatcacc gcgtcgaaat taccgaggga atcctggcag atgaatgtgc cgccctgctg
     1741 tgctatttct ttcggatgcc tagacaggtg ttcaatgctc agaagaaggc ccagagctcc
     1801 accgactccg gaggatctag cggaggctcc tctggctctg agacacctgg cacaagcgag
     1861 agcgcaacac ctgaaagcag cgggggcagc agcggggggt cacacgagaa ccagcagaag
     1921 cagaactaca tcctgggcct ggccatcgga atcaccagcg tcggctacgg actgatcgat
     1981 agcaagacaa gagaagtgat cgacgccggc gttagactct ttccagaagc tgatagcgag
     2041 aacaactcca accgcagaag caagcggggc gccagacggt taaaacggag aagaatccac
     2101 cggctgaacc gggtcaaaga cctgctcgct gattaccaga tgatcgatct taacaatgtt
     2161 cctaagagca ccgaccccta caccatcaga gtgaagggcc tccgggagcc tctgacaaaa
     2221 gaagaattcg ccatcgccct cctgcatatc gctaagagaa gaggcctgca caacatcagt
     2281 gtgtccatgg gcgacgaaga gcaggacaat gaactgagca ccaagcagca gctgcaaaag
     2341 aatgcccagc aactgcagga caagtatgtg tgcgaactgc agttagaacg gctgaccaac
     2401 atcaacaagg tcagaggcga gaagaacaga tttaagacag aggactttgt gaaagaagtg
     2461 aaacagctgt gcgaaaccca gagacagtac cacaacatcg acgaccaatt catccagcag
     2521 tacatcgacc tggtgtctac aagacgggag tacttcgagg gccccggcaa cggctctcca
     2581 tacggctggg acggcgacct gctgaagtgg tacgagaagc tgatgggcag atgcacctat
     2641 ttccccgaag aactgaggtc cgtgaagtac gcctacagcg ccgacctctt caacgccctg
     2701 aacgacctga acaacctcgt tgtgaccagg gatgacaatc caaagcttga gtactacgag
     2761 aagtaccaca ttattgagaa cgtgttcaag caaaagaaga atcccacact caaacaaatc
     2821 gccaaagaga tcggcgtgca agattacgac atccggggct atagaatcac aaagagcggc
     2881 aaacctcagt tcacctcttt taagctgtat cacgacctga agaacatctt cgagcaggcc
     2941 aaatacctgg aagatgtgga aatgctggac gagatcgcca agatcctgac catctaccag
     3001 gatgagatta gcatcaagaa agccctggac cagctgcccg aactgctgac agagagcgag
     3061 aaatctcaga tcgcacagct caccggctat acaggcaccc acagactgag cctgaagtgc
     3121 atccacattg tgatcgacga gctgtgggag agccccgaga accagatgga aatctttacc
     3181 agactgaatc tgaaacctaa gaaggtggaa atgagcgaga tcgacagcat acccaccacc
     3241 ctggtcgacg agttcatcct ctcacctgtg gtgaagcggg ccttcatcca gagcatcaag
     3301 gtaatcaacg cagtgatcaa tcggttcggc ctgccagagg acatcatcat cgagctggcc
     3361 agagaaaaga atagcaagga tcggagaaag ttcattaaca agctgcagaa acaaaatgag
     3421 gccacaagaa agaaaatcga acagctgctg gccaagtacg gcaacaccaa tgccaagtac
     3481 atgatcgaga agatcaagct gcacgacatg caggagggca agtgcctgta cagcctggag
     3541 gctattcctc tggaagacct gctgagcaac ccgacacact acgaagttga ccacattatc
     3601 cccagatctg tgagctttga caacagcctg aacaacaaag tgctggtgaa acaaagcgaa
     3661 aacagcaaga agggcaatcg caccccttac cagtacctga gcagcaacga gtctaagatt
     3721 agctacaacc agtttaagca gcacatcctg aacctgagca aggccaagga cagaatcagc
     3781 aagaaaaaaa gagatatgct gctggaagag agagatatca acaagttcga agtgcagaag
     3841 gaattcatta accggaacct ggtggataca cggtacgcca ccagagaact gtctaacctg
     3901 ctgaagacct acttcagcac ccatgactac gccgtgaagg tgaagaccat caacggcggc
     3961 ttcactaacc acctgaggaa ggtgtgggat ttcaagaagc acagaaacca cggctacaag
     4021 caccacgccg aagatgccct ggtgatcgcc aacgccgact tcctgtttaa gacacataag
     4081 gccctgcgga gaaccgataa gatcctggaa caacctggcc tggaagtgaa tgatacaacc
     4141 gtgaaagtgg acaccgagga aaaataccag gagctgttcg agacacctaa gcaagtgaag
     4201 aacatcaagc agttccggga cttcaagtac agccaccgag tggacaagaa gcctaaccgg
     4261 cagcttatca acgacacact gtactccacc agagagattg atggcgaaac ctacgtggtg
     4321 cagaccctta aggatctgta cgccaaggac aacgagaaag tgaagaagct gttcaccgaa
     4381 agacctcaga agatcctgat gtaccagcac gaccctaaga ccttcgagaa actgatgaca
     4441 atcctgaacc agtacgctga ggccaagaac cctctggctg cttattacga ggacaaaggc
     4501 gagtacgtga ccaagtacgc caagaaaggc aatggacctg ccatccacaa gatcaagtat
     4561 atcgataaga agcttggatc ttacctggat gttagcaaca agtatcctga gacacagaac
     4621 aagcttgtga agctgtccct gaagagcttt agattcgaca tctacaagtg tgaacagggc
     4681 tacaagatgg tgtccatcgg atacctggac gtgctgaaga aagataacta ctactacatc
     4741 cctaaggaca agtacgaggc cgagaagcag aaaaagaaga tcaaggaatc tgatcttttt
     4801 gtgggcagct tctactacaa cgacctcatc atgtacgagg atgaactgtt cagagtgata
     4861 ggagtgaaca gcgacatcaa caatctggtt gagctaaaca tggtcgacat tacctacaag
     4921 gacttctgcg aggtgaacaa cgtgacaggc gagaaaagaa tcaaaaagac tatcggcaag
     4981 cgcgtggtcc tgatcgagaa gtacaccaca gatattctag gcaacctgta caagactccc
     5041 ctgcctaaga agccccagct tatcttcaag cggggagaac tgtctggcgg ctcaaaaaga
     5101 accgccgacg gcagcgaatt cgagcccaag aagaagagga aagtcggaag cggagctact
     5161 aacttcagcc tgctgaagca ggctggagac gtggaggaga accctggacc tatggtgagc
     5221 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     5281 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     5341 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     5401 accctgacct atggagtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     5461 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     5521 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     5581 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     5641 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     5701 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     5761 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     5821 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     5881 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc tggtggttct
     5941 cccaagaaga agaggaaagt ctaaccggtc atcatcacca tcaccattga gtttaaaccc
     6001 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     6061 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     6121 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     6181 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     6241 cttctgaggc ggaaagaacc agctggggct cgataccgtc gacctctagc tagagcttgg
     6301 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6361 acatacgagc cggaagcata aagtgtaaag cctagggtgc ctaatgagtg agctaactca
     6421 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6481 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6541 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6601 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6661 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6721 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6781 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6841 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6901 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6961 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7021 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     7081 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7141 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7201 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7261 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7321 ctacggggtc tgacactcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7381 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7441 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7501 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7561 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7621 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7681 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7741 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7801 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7861 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7921 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7981 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     8041 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     8101 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     8161 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     8221 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     8281 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8341 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8401 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8461 ctgacgtcga cggatcggga gatcgatctc ccgatcccct agggtcgact ctcagtacaa
     8521 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     8581 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     8641 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
     8701 cgcgtt
//
