LOCUS       sequence-344939-        8028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes human codon optimized HvsCas7-11 protein with C-terminal NES
            tag for expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8028)
  AUTHORS   Ozcan A, Krajeski R, Ioannidi E, Lee B, Gardner A, Makarova KS, 
            Koonin EV, Abudayyeh OO, Gootenberg JS
  TITLE     Programmable RNA targeting with the single-protein CRISPR effector 
            Cas7-11.
  JOURNAL   Nature. 2021 Sep 6. pii: 10.1038/s41586-021-03886-5. doi: 
            10.1038/s41586-021-03886-5.
  PUBMED    34489594
REFERENCE   2  (bases 1 to 8028)
  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: "Nature. 
            2021 Sep 6. pii: 10.1038/s41586-021-03886-5. doi: 
            10.1038/s41586-021-03886-5."
COMMENT     Sequence Label: pDF0352 HvsCas7-11 with C-term Nes mammalian
            e...
FEATURES             Location/Qualifiers
     source          1..8028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     860..879
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        860..878
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5526..5543
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5566..5583)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5572..5796
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5867..5883)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5867..5883)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5880..5902)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5891..5907
                     /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(5915..5921,5922..5939,5940..5945))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5960..5981
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6098..6115)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6269..6857)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6349..6368)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7028..7819,7820..7888))
                     /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     7651..7670
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7889..7993)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gatcgggaga tcgatctccc gatcccctag ggtctactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
      841 gctagagatc cgcggccgct aatacgactc actataggga gagccgccac catgaagtcc
      901 atccctataa cactgacttt tctggaaccc tacaggatac tgccctgggc cgaaaagggg
      961 aaacgagata aaaaggagta cctgcgaggt gccaattatg ttcggttgca taaggataaa
     1021 aatggaaagt tcaagcccta cataacaggt acattaatcc ggagcgccgt gctgtcagca
     1081 atcgagatgt tgctggacat cacaaacggc gaatggaacg gcaaagaatg ttgcttggca
     1141 aaattccaca ctgagggtga gaagcccagc tttctgcgga agaagcccat ttacatccgc
     1201 gctgaaaaag acgagatttg cacctcccgg gaaacggcat gtccactgtg cctcatactc
     1261 gggcggtttg acaaagcgga aaaaaaggag aaagataaag aaaaatttga tgtgcacttc
     1321 agcaatctta atttatatag ttctaaggag ttcagtacta tcgaagagct ggctcctaag
     1381 cgggcactca atcggattga gcagtacact ggaaaggccc aagattactt cacagtatat
     1441 gaagccctga ataaagagtt ttggacattt aagggacgga tccggattaa agaagatatc
     1501 tacgacaagg tgacagatct gcttttttct gcactgaggt gcgttgagaa aattgccggc
     1561 gcactttgtc ggattgagat tgataaagaa cccagccagc agaaggggtt cgtgaaacgc
     1621 cagcttagta agcaggccaa agaagatatt gaaaagatat ttcaggtggt taaagacgct
     1681 cagaagctca gactgctgag tgactgtttt agagagttaa caagaatggc aaacaaagat
     1741 gagctggctt tgccattagg cccagaggac gacggacatt atctgtggga taagattaaa
     1801 gttgagggaa agacactgcg catattcttg cggaattgtt ttagtcaata taaggacaat
     1861 tggttatgtt tttgcgacga agccagtaaa aagggctacc agaagtacag agagaagaga
     1921 cacaaactga cggatagaga actgccaact gccactccaa aacacttcgc cgaaaaaaaa
     1981 gatccacaga tatctccaat ctatattgac aaggatgaca aggtttatga gtggatcata
     2041 gtcggcaggc tcatcgctca aacacctttt catttcggcg atgaagagaa agccgagggc
     2101 gcaattctcc tcactccaga taatcggttc aggctgccac gaacagcact gaggggaatc
     2161 ctgagaaggg acttaaaact cgccggtgca tcagcctgtg aggtggaagt ggggcggagt
     2221 gagccatgcc cttgtgacgt gtgtaagatt atgcgccgcg tgacactgtt ggataccgtg
     2281 tctgaggacc ttcgggattt cctgccagag ttacggaaaa gaattcggat caatccgcag
     2341 agcggcactg tggcagaggg cgcgctgttc gacacagaag tgggacccga gggtttgtcc
     2401 tttcccttcg tgctcagata caagtgcgag aaactgcccg acagcttgac tacagtcctg
     2461 tgttggtggc aggaagggtt ggctttcctt tccggagaga gtgcgacagg gaaaggtcgg
     2521 tttagattag agattaacgg cgcttttgta tgggatctcc agaagggctt gtttaactat
     2581 ataaaaaacc acggatttcg gggcgaagaa aggttattcc tggaaggtaa tgaggccgaa
     2641 ctcgaaaaaa tggggatcca gatcaatacc gaattgttgc aaccagaaat gatcaagaag
     2701 gaaaaaaatt ttaccgattt cccctatgac ttaattaagt accagctgaa catttccagc
     2761 cctctccttc ttaacgaccc aattagggct attgctttgt acgaggggga agggaaagcc
     2821 ccagacgcag tgttcttcaa aaagtacgtc tttgaaaatg gaaagattga ggaaaagcca
     2881 tgttttaaag ctgagtccat caggggaatc ttcaggaccg cagtgggacg gatcaagaat
     2941 gtgttaacta agaatcacga ggattgcatt tgtgtactgt gccacctgtt cggcaatgtg
     3001 catgaaactg ggagattaaa gtttgaagat cttaaaatcg tctccggcca ggaggagaaa
     3061 ttttttgacc acgtggcaat tgaccgcttc ttggggggcg caaaagagaa atacaagttc
     3121 gacgataaac ccatcatcgg ggcacccgac acccctatcg tcctggaggg caagatctgg
     3181 gtcaagaaag atatcaatga tgaagcaaaa gagacactga gccaggcctt ctcagatata
     3241 aatactggca tttactacct gggggcaaac ggctctatcg gttacggctg gatcgaagag
     3301 gttaaggctc tcaaggctcc ctcctggctt aagataaagg aaaagcctaa tttcgagaag
     3361 gatactagtt tgaatattag tgccattatg aatgaattca agaaagacat tcagacactg
     3421 aatttggata agacatatct gccgtatggc ttcctcaaat tactcgagaa ggttaaacgt
     3481 acctcaagtc ctattactca cgagcgcttc tacgaaaacc acttaactgg attcatcgag
     3541 tgtagtctga aagtcctgtc tcctctgatt attcccgaca ccgaaacacc cgagaaggaa
     3601 gaaaatggac acaagtacta tcacttcctc aaaatagaca acaagcctat cattccgggc
     3661 gccgagatta gaggtgctgt gtcatctatt tacgaggctc ttactaattc atgttttcgt
     3721 gtgttcggtg aaaagaaagt cctgtcctgg cggatggaag gtaaggatgc aaaggaattt
     3781 atgcctggca gagtgtccaa gaagaaaggt aagttgtaca tggtaaagat gcaggccctg
     3841 cgcttaccgg tatacgacaa cccggccctg gccaacgaaa ttcggtctgg gtcaatctac
     3901 gagaagtaca agaactccaa agtggaaatc attttctttc aaacagttga agggattcga
     3961 aagttcctcc gcggaaattt caataatgtc gagtggaaga aagtattggt cactggtatt
     4021 gaccctctcg caattttgcc ctcccagaaa atccccggca atgacaagtg ggttaagaac
     4081 ctccagagca aaatttcccc cgtccggggg tacttcaaat ttaccgggcc aaacaagatc
     4141 gaaacaaagc gcagggagga agagaaggat gagaaattaa gaaccaaagc taacaaggtc
     4201 tcttgcctgc agaaggacaa atggtatgaa gctatgcaca accacgtgga atataagcag
     4261 gattatacac cccctaacag ccctaaaacc gaacccctgg aacggcctag aaatatccca
     4321 tgcttcgtgt gcagcgataa ggagaagata tacagaatga ccaagcggtg cgagagggtc
     4381 tttgtgagcc tgggggagaa cgcacctaag tatgagatcc ccatctcagc tattaagcgc
     4441 tatgaagtga tcctgtcagc ctaccgcgag aactgggaga gaaataaaac cccagaactg
     4501 tttcgcacca gattgccagg ggatgggcgg actttgaatg aagatgatct ggtgtatttt
     4561 agggcagacg aaaacgagaa agtgaaggat atcatccccg tttgtattag taggatcgtc
     4621 gacgaggtcc ctctcatcaa aaggctgtcc caagagctgt ggccctgcgt gctggctgaa
     4681 tgccccctgc tgggctttga gtgcaagaag tgcgagctcg agggactgcc tgagaagatc
     4741 tggttccgta taaacaagga tgggctctgt cctgcatgca gactcttcgg aactcagatc
     4801 tacaaatcca gagtgcggtt ttccttcgcc tacgctaaga attggaagtt ctacgacgga
     4861 tacattacac tccctcggct ggagtctcca cgcgcaacct ggctgattct taaagagaag
     4921 gataagcact acatcaagta caaggtttgt ggtagaaagt tctacctgca caactcaaca
     4981 tatgaggata tcatcaacaa tagcaagaag gagaaggaga agaagaccga gaataatgcc
     5041 tcttttgagg tgctcaagga gggtgaattc acattcaagg tttacttcga aaacttagag
     5101 aactgggagt taggtctgct tttgctgtcc ttaacagggc tcggcgaggc tatcaagata
     5161 ggccacgcta aacctctcgg tttcgggtct gtgaagatag aggccaaaaa gatctacttt
     5221 cgagaagagg ccgggaagtt ccacccttgc gagaaagcag acgaatacct taagaagggc
     5281 ttgaacaagc tgaccagctg gttcggtaag aacgaaatca atgagcacat gcgaaactta
     5341 ctcttgttca tgacttacta tcagaacctt cccaaggtta agtaccctga ttttgacgga
     5401 tacgcaaaat ggcggtgcag ctacgttgaa caggacaagg tggaatactt tcagaaccgc
     5461 tggattgtgg cctctggcgg ctcactctat ccagagaggc tgcgccgtat tctgacataa
     5521 ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt
     5581 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     5641 ccactcccac tgtcctttcc taataaaatg agaaaattgc atcgcattgt ctgagtaggt
     5701 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     5761 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     5821 ggggctcgat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5881 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5941 gtaaagccta gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6001 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6061 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6121 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6181 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6241 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6301 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6361 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6421 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6481 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6541 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6601 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6661 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6721 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6781 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6841 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac actcagtgga
     6901 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6961 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7021 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7081 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7141 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7201 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7261 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7321 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7381 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7441 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7501 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7561 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7621 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7681 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7741 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7801 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7861 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7921 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7981 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacg
//
