LOCUS       sequence-418035-        7118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SynCas.v35 for analysis of RNA knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7118)
  AUTHORS   Brogan DJ, Dalla Benetta E, Wang T, Lin CP, Chen F, Li H, Lin C, 
            Komives EA, Akbari OS
  TITLE     Synthetic type III-E CRISPR-Cas effectors for programmable 
            RNA-targeting
  JOURNAL   bioRxiv 2024.02.23.581838; doi: 
            https://doi.org/10.1101/2024.02.23.581838
REFERENCE   2  (bases 1 to 7118)
  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: "bioRxiv 
            2024.02.23.581838; doi: https://doi.org/10.1101/2024.02.23.581838"
COMMENT     Sequence Label: CMV-SynCas.v35 mammalian expression
FEATURES             Location/Qualifiers
     source          1..7118
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     536..555
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(774..878)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(957..976)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1148..1527
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1528..1731
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1682..1702
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5529..5546
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5569..5586)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5575..5799
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5870..5886)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5870..5886)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5883..5905)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5894..5910
                     /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(5918..5924,5925..5942,5943..5948))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5963..5984
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6101..6118)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6272..6860)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6352..6371)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7031..7118,1..704,705..773))
                     /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"
ORIGIN
        1 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
       61 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
      121 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
      181 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
      241 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
      301 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
      361 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
      421 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
      481 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
      541 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
      601 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
      661 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
      721 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
      781 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
      841 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
      901 cctgacgtcg acggatcggg agatcgatct cccgatcccc tagggtcgac tctcagtaca
      961 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     1021 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     1081 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     1141 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     1201 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     1261 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     1321 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     1381 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     1441 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     1501 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     1561 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     1621 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     1681 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
     1741 aaccgtcaga tccgctagag atccgcggcc gcatgacgac tactatgaag atttcaattg
     1801 aattcctcga gccgtttcgg atgaccaaat ggcaggaaag cactaggcga aacaagaaca
     1861 acaaggaatt tgttcgcggc caagcttttg caagatggca tagaaataag aaggataaca
     1921 caaaagggcg gccttacata actgggacat tgctgcggtc cgcagtaatt agaagcgcgg
     1981 aaaacctgct gacattgagt gatgggaaaa ttagcgaaaa gacatgttgt cctgggaaat
     2041 ttgatacgga agataaagac aggttgctgc aattgaggca gcggtccacc cttcgttgga
     2101 ccgataagaa cccctgtcca gataacgctg aaacttactg cccgttttgc gaacttctcg
     2161 ggagatccgg taacgatgga aagaaggctg agaagaagga ttggcgcttt cgtattcatt
     2221 ttggcaatct ttcccttcca gggaagcccg atttcgacgg gccaaaagct ataggcagcc
     2281 aacgggtact taacagggtc gatttcaaat ccggtaaggc ccatgacttc tttaaggctt
     2341 acgaagtcga ccatactagg tttccccgct tcgaagggga gattaccata gataataaag
     2401 tcagcgctga agccaggaaa ctgctctgtg attctctcaa gtttacggat cggctgtgtg
     2461 gagctctgtg cgtaatcagg ttcgatgaat atacaccagc agccgatagt gggaaacaaa
     2521 ccgaaaatgt ccaagcagaa ccgaatgcta atctcgctga aaagaccgca gagcaaatta
     2581 ttagcattct ggacgataat aagaaaaccg aatatacccg cctgcttgct gatgctattc
     2641 gttcactccg ccgctctagt aaacttgtcg ctggcttgcc aaaggatcat gacgggaagg
     2701 atgaccatta cctctgggat atcggcaaga agaagaaaga cgaaaactct gtcactatta
     2761 ggcaaatcct tacgacctca gcagacacca aggaactcaa gaatgccgga aaatggagag
     2821 aattctgcga gaagctgggt gaagcgctgt acctcaagag taaagacatg agtggcggcc
     2881 tgaaaattac tcggagaata ctgggcgatg ctgaattcca tggaaagccc gatcggcttg
     2941 aaaagagccg gtctgtgtca atcggatctg tgttgtatga gtggatcata gtcggcaggc
     3001 tcatcgctca aacacctttt catttcggcg atgaagagaa agccgagggc gcaattctcc
     3061 tcactccaga taatcggttc aggctgccac gaacagcact gaggggaatc ctgagaaggg
     3121 acttaaaact cgccggtgca tcagcctgtg aggtggaagt ggggcggagt gagccatgcc
     3181 cttgtgacgt gtgtaagatt atgcgccgcg tgacactgtt ggataccgtg tctgaggacc
     3241 ttcgggattt cctgccagag ttacggaaaa gaattcggat caatccgcag agcggcactg
     3301 tggcagaggg cgcgctgttc gacacagaag tgggacccga gggtttgtcc tttcccttcg
     3361 tgctcagata caagtgcgag aaactgcccg acagcttgac tacagtcctg tgttggtggc
     3421 aggaagggtt ggctttcctt tccggagaga gtgcgacagg gaaaggtcgg tttagattag
     3481 agattaacgg cgcttttgta tgggatctca gcgacgagaa tcaaaggaat gactatctta
     3541 agaattgggg atggcgtgac gagaaggggc tcgaggaact gaagaaacga ctgaactcag
     3601 gtctgccaga gcccggtaat tatagggatc caaaatggga cttaattaag taccagctga
     3661 acatttccag ccctctcctt cttaacgacc caattagggc tattgctttg tacgaggggg
     3721 aagggaaagc cccagacgca gtgttcttca aaaagtacgt ctttgaaaat ggaaagattg
     3781 aggaaaagcc atgttttaaa gctgagtcca tcaggggaat cttcaggacc gcagtgggac
     3841 ggatcaagaa tgtgttaact aagaatcacg aggattgcat ttgtgtactg tgccacctgt
     3901 tcggcaatgt gcatgaaact gggagattaa agtttgaaga tcttaaaatc gtctccggcc
     3961 aggaggagaa attttttgac cacgtggcaa ttgaccgctt cttggggggc gcaaaagaga
     4021 aatacaagtt cgacgataaa cccatcatcg gggcacccga cacccctatc gtcctggagg
     4081 gcaagatctg ggtcaagaaa gatatcaatg atgaagcaaa agagacactg agccaggcct
     4141 tctcagatat aaatactggc atttactacc tgggggcaaa cggctctatc ggttacggct
     4201 ggatcgaaga ggttaaggct ctcaaggatg ataagaggat ttctcgcctc atgaatcctg
     4261 cctttgatga gacagatgta gccgttccag aaaagcccaa aactgatgcc gaggttcgca
     4321 tcgaggcaga gaaagtatat tacccacact atttcgtcga accccataag aaggtggaac
     4381 gcgaggagaa accctgtggt catcaaaagt tccacgaggg gcgactgact ggtaaaattg
     4441 agtgtacact gaaaacgctg agccctttaa taataccgga cagtgaggtc gttacggaga
     4501 acggaaacgg ccacaagtct taccagttct tcagattaaa taatgaaatc atcattccgg
     4561 gaagcgaaat tcggggtatg atcagttccg tgtacgaagc acttactaac tcttgcttcc
     4621 gtgtcgtgga cgacatcatc ccggtggtga tctcccgtag cgtgtcaccg taccagatcg
     4681 ctcagaagct gcccagtccg gatctgcgtc cttgcgagcg tgagattctt gaggactcag
     4741 aaattggatc tttagacaag tgttcagaga aacgcctttt ccgcctgcat gaagacggtc
     4801 tttgcccatc gtgtcgtctg ttcggcacta cacattataa gggacgcgtt cggttcggct
     4861 tcgcaaagca cgagggcggc gaaaagtggc tcatggacga caagactaag gacgagtatc
     4921 tgactttacc gttgttggaa cggccgcgcc caacatggag catgccaagc gacgaagacg
     4981 acgtaccggg acggaaattc tacgtacacc acaatggttg ggagaaagtt aagcaagata
     5041 gtttagataa taaaacgagc aaaaccgaga ataatcgaag cgtggaacca ctcgataagg
     5101 ataacctgtt taaattcgac gtgttcttcg agaacctgga accctgggaa cttggtttgc
     5161 ttctctactc attagaattg gaagaggggt tagcccacaa gtttggaatg gcaaaagcgt
     5221 tcggcttcgg ctcgacaaag attgacgccg acaaaatact gttacgtgaa gaggccggga
     5281 agttccaccc ttgcgagaaa gcagacgaat accttaagaa gggcttgaac aagctgacca
     5341 gctggttcgg taagaacgaa atcaatgagc acatgcgaaa cttactcttg ttcatgactt
     5401 actatcagaa ccttcccaag gttaagtacc ctgattttga cggatacgca aaatggcggt
     5461 gcagctacgt tgaacaggac aaggtggaat actttcagaa ccgctggatt gtggcctctt
     5521 aaaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
     5581 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     5641 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     5701 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     5761 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     5821 gctggggctc gataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5881 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5941 agtgtaaagc ctaggatgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6001 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6061 cgggaagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6121 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6181 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6241 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6301 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6361 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6421 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6481 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6541 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6601 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6661 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6721 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6781 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6841 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacactcagt
     6901 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6961 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7021 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7081 gttcatccat agttgcctga ctccccgtcg tgtagata
//
