LOCUS       sequence-344934-        7986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes human codon optimized HvsCas7-11 protein alone for 
            expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7986)
  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 7986)
  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: pDF0317 HvsCas7-11 mammalian expression
FEATURES             Location/Qualifiers
     source          1..7986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(70..89)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        261..640
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        641..844
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     795..815
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     886..905
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        886..904
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5510..5527
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5550..5567)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5556..5780
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5851..5867)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5851..5867)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5864..5886)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5875..5891
                     /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(5899..5905,5906..5923,5924..5929))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5944..5965
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6082..6099)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6253..6841)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6333..6352)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7012..7803,7804..7872))
                     /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     7635..7654
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7873..7977)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgaaaagtg ccacctgacg tcgacggatc gggagatcga tctcccgatc ccctagggtc
       61 tactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
      121 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
      181 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
      241 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
      301 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      361 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      421 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      481 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      541 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      601 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      661 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      721 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      781 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      841 agctggttta gtgaaccgtc agatccgcta gagatccgcg gccgctaata cgactcacta
      901 tagggagagc cgccaccatg aagtccatcc ctataacact gacttttctg gaaccctaca
      961 ggatactgcc ctgggccgaa aaggggaaac gagataaaaa ggagtacctg cgaggtgcca
     1021 attatgttcg gttgcataag gataaaaatg gaaagttcaa gccctacata acaggtacat
     1081 taatccggag cgccgtgctg tcagcaatcg agatgttgct ggacatcaca aacggcgaat
     1141 ggaacggcaa agaatgttgc ttggcaaaat tccacactga gggtgagaag cccagctttc
     1201 tgcggaagaa gcccatttac atccgcgctg aaaaagacga gatttgcacc tcccgggaaa
     1261 cggcatgtcc actgtgcctc atactcgggc ggtttgacaa agcggaaaaa aaggagaaag
     1321 ataaagaaaa atttgatgtg cacttcagca atcttaattt atatagttct aaggagttca
     1381 gtactatcga agagctggct cctaagcggg cactcaatcg gattgagcag tacactggaa
     1441 aggcccaaga ttacttcaca gtatatgaag ccctgaataa agagttttgg acatttaagg
     1501 gacggatccg gattaaagaa gatatctacg acaaggtgac agatctgctt ttttctgcac
     1561 tgaggtgcgt tgagaaaatt gccggcgcac tttgtcggat tgagattgat aaagaaccca
     1621 gccagcagaa ggggttcgtg aaacgccagc ttagtaagca ggccaaagaa gatattgaaa
     1681 agatatttca ggtggttaaa gacgctcaga agctcagact gctgagtgac tgttttagag
     1741 agttaacaag aatggcaaac aaagatgagc tggctttgcc attaggccca gaggacgacg
     1801 gacattatct gtgggataag attaaagttg agggaaagac actgcgcata ttcttgcgga
     1861 attgttttag tcaatataag gacaattggt tatgtttttg cgacgaagcc agtaaaaagg
     1921 gctaccagaa gtacagagag aagagacaca aactgacgga tagagaactg ccaactgcca
     1981 ctccaaaaca cttcgccgaa aaaaaagatc cacagatatc tccaatctat attgacaagg
     2041 atgacaaggt ttatgagtgg atcatagtcg gcaggctcat cgctcaaaca ccttttcatt
     2101 tcggcgatga agagaaagcc gagggcgcaa ttctcctcac tccagataat cggttcaggc
     2161 tgccacgaac agcactgagg ggaatcctga gaagggactt aaaactcgcc ggtgcatcag
     2221 cctgtgaggt ggaagtgggg cggagtgagc catgcccttg tgacgtgtgt aagattatgc
     2281 gccgcgtgac actgttggat accgtgtctg aggaccttcg ggatttcctg ccagagttac
     2341 ggaaaagaat tcggatcaat ccgcagagcg gcactgtggc agagggcgcg ctgttcgaca
     2401 cagaagtggg acccgagggt ttgtcctttc ccttcgtgct cagatacaag tgcgagaaac
     2461 tgcccgacag cttgactaca gtcctgtgtt ggtggcagga agggttggct ttcctttccg
     2521 gagagagtgc gacagggaaa ggtcggttta gattagagat taacggcgct tttgtatggg
     2581 atctccagaa gggcttgttt aactatataa aaaaccacgg atttcggggc gaagaaaggt
     2641 tattcctgga aggtaatgag gccgaactcg aaaaaatggg gatccagatc aataccgaat
     2701 tgttgcaacc agaaatgatc aagaaggaaa aaaattttac cgatttcccc tatgacttaa
     2761 ttaagtacca gctgaacatt tccagccctc tccttcttaa cgacccaatt agggctattg
     2821 ctttgtacga gggggaaggg aaagccccag acgcagtgtt cttcaaaaag tacgtctttg
     2881 aaaatggaaa gattgaggaa aagccatgtt ttaaagctga gtccatcagg ggaatcttca
     2941 ggaccgcagt gggacggatc aagaatgtgt taactaagaa tcacgaggat tgcatttgtg
     3001 tactgtgcca cctgttcggc aatgtgcatg aaactgggag attaaagttt gaagatctta
     3061 aaatcgtctc cggccaggag gagaaatttt ttgaccacgt ggcaattgac cgcttcttgg
     3121 ggggcgcaaa agagaaatac aagttcgacg ataaacccat catcggggca cccgacaccc
     3181 ctatcgtcct ggagggcaag atctgggtca agaaagatat caatgatgaa gcaaaagaga
     3241 cactgagcca ggccttctca gatataaata ctggcattta ctacctgggg gcaaacggct
     3301 ctatcggtta cggctggatc gaagaggtta aggctctcaa ggctccctcc tggcttaaga
     3361 taaaggaaaa gcctaatttc gagaaggata ctagtttgaa tattagtgcc attatgaatg
     3421 aattcaagaa agacattcag acactgaatt tggataagac atatctgccg tatggcttcc
     3481 tcaaattact cgagaaggtt aaacgtacct caagtcctat tactcacgag cgcttctacg
     3541 aaaaccactt aactggattc atcgagtgta gtctgaaagt cctgtctcct ctgattattc
     3601 ccgacaccga aacacccgag aaggaagaaa atggacacaa gtactatcac ttcctcaaaa
     3661 tagacaacaa gcctatcatt ccgggcgccg agattagagg tgctgtgtca tctatttacg
     3721 aggctcttac taattcatgt tttcgtgtgt tcggtgaaaa gaaagtcctg tcctggcgga
     3781 tggaaggtaa ggatgcaaag gaatttatgc ctggcagagt gtccaagaag aaaggtaagt
     3841 tgtacatggt aaagatgcag gccctgcgct taccggtata cgacaacccg gccctggcca
     3901 acgaaattcg gtctgggtca atctacgaga agtacaagaa ctccaaagtg gaaatcattt
     3961 tctttcaaac agttgaaggg attcgaaagt tcctccgcgg aaatttcaat aatgtcgagt
     4021 ggaagaaagt attggtcact ggtattgacc ctctcgcaat tttgccctcc cagaaaatcc
     4081 ccggcaatga caagtgggtt aagaacctcc agagcaaaat ttcccccgtc cgggggtact
     4141 tcaaatttac cgggccaaac aagatcgaaa caaagcgcag ggaggaagag aaggatgaga
     4201 aattaagaac caaagctaac aaggtctctt gcctgcagaa ggacaaatgg tatgaagcta
     4261 tgcacaacca cgtggaatat aagcaggatt atacaccccc taacagccct aaaaccgaac
     4321 ccctggaacg gcctagaaat atcccatgct tcgtgtgcag cgataaggag aagatataca
     4381 gaatgaccaa gcggtgcgag agggtctttg tgagcctggg ggagaacgca cctaagtatg
     4441 agatccccat ctcagctatt aagcgctatg aagtgatcct gtcagcctac cgcgagaact
     4501 gggagagaaa taaaacccca gaactgtttc gcaccagatt gccaggggat gggcggactt
     4561 tgaatgaaga tgatctggtg tattttaggg cagacgaaaa cgagaaagtg aaggatatca
     4621 tccccgtttg tattagtagg atcgtcgacg aggtccctct catcaaaagg ctgtcccaag
     4681 agctgtggcc ctgcgtgctg gctgaatgcc ccctgctggg ctttgagtgc aagaagtgcg
     4741 agctcgaggg actgcctgag aagatctggt tccgtataaa caaggatggg ctctgtcctg
     4801 catgcagact cttcggaact cagatctaca aatccagagt gcggttttcc ttcgcctacg
     4861 ctaagaattg gaagttctac gacggataca ttacactccc tcggctggag tctccacgcg
     4921 caacctggct gattcttaaa gagaaggata agcactacat caagtacaag gtttgtggta
     4981 gaaagttcta cctgcacaac tcaacatatg aggatatcat caacaatagc aagaaggaga
     5041 aggagaagaa gaccgagaat aatgcctctt ttgaggtgct caaggagggt gaattcacat
     5101 tcaaggttta cttcgaaaac ttagagaact gggagttagg tctgcttttg ctgtccttaa
     5161 cagggctcgg cgaggctatc aagataggcc acgctaaacc tctcggtttc gggtctgtga
     5221 agatagaggc caaaaagatc tactttcgag aagaggccgg gaagttccac ccttgcgaga
     5281 aagcagacga ataccttaag aagggcttga acaagctgac cagctggttc ggtaagaacg
     5341 aaatcaatga gcacatgcga aacttactct tgttcatgac ttactatcag aaccttccca
     5401 aggttaagta ccctgatttt gacggatacg caaaatggcg gtgcagctac gttgaacagg
     5461 acaaggtgga atactttcag aaccgctgga ttgtggcctc ttaaccggtc atcatcacca
     5521 tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     5581 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     5641 ttcctaataa aatgagaaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     5701 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     5761 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct cgataccgtc
     5821 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5881 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctagggtgc
     5941 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6001 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6061 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6121 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6181 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6241 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6301 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6361 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6421 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6481 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6541 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6601 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6661 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6721 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6781 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6841 agaagatcct ttgatctttt ctacggggtc tgacactcag tggaacgaaa actcacgtta
     6901 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6961 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7021 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7081 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7141 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7201 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7261 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7321 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7381 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7441 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7501 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7561 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7621 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7681 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7741 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7801 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7861 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7921 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7981 atttcc
//
