LOCUS       sequence-418034-        7229 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SynCas.v47 for analysis of RNA knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7229)
  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 7229)
  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.v47 mammalian expression
FEATURES             Location/Qualifiers
     source          1..7229
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     539..558
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(777..881)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(960..979)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1151..1530
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1531..1734
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1685..1705
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5643..5660
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5683..5700)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5689..5913
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5984..6000)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5984..6000)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5997..6019)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6008..6024
                     /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(6032..6038,6039..6056,6057..6062))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6077..6098
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6215..6232)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6386..6974)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6466..6485)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7145..7229,1..707,708..776))
                     /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 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
       61 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
      121 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
      181 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
      241 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
      301 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
      361 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
      421 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
      481 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
      541 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
      601 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
      661 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
      721 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
      781 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
      841 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
      901 ccacctgacg tcgacggatc gggagatcga tctcccgatc ccctagggtc gactctcagt
      961 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     1021 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     1081 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     1141 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     1201 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     1261 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     1321 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     1381 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     1441 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1501 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     1561 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     1621 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     1681 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta
     1741 gtgaaccgtc agatccgcta gagatccgcg gccgcatgac gactactatg aagatttcaa
     1801 ttgaattcct cgagccgttt cggatgacca aatggcagga aagcactagg cgaaacaaga
     1861 acaacaagga atttgttcgc ggccaagctt ttgcaagatg gcatagaaat aagaaggata
     1921 acacaaaagg gcggccttac ataactggga cattgctgcg gtccgcagta attagaagcg
     1981 cggaaaacct gctgacattg agtgatggga aaattagcga aaagacatgt tgtcctggga
     2041 aatttgatac ggaagataaa gacaggttgc tgcaattgag gcagcggtcc acccttcgtt
     2101 ggaccgataa gaacccctgt ccagataacg ctgaaactta ctgcccgttt tgcgaacttc
     2161 tcgggagatc cggtaacgat ggaaagaagg ctgagaagaa ggattggcgc tttcgtattc
     2221 attttggcaa tctttccctt ccagggaagc ccgatttcga cgggccaaaa gctataggca
     2281 gccaacgggt acttaacagg gtcgatttca aatccggtaa ggcccatgac ttctttaagg
     2341 cttacgaagt cgaccatact aggtttcccc gcttcgaagg ggagattacc atagataata
     2401 aagtcagcgc tgaagccagg aaactgctct gtgattctct caagtttacg gatcggctgt
     2461 gtggagctct gtgcgtaatc aggttcgatg aatatacacc agcagccgat agtgggaaac
     2521 aaaccgaaaa tgtccaagca gaaccgaatg ctgcggttaa gaaggctgcg ggagatctgg
     2581 ttaacagcgc gttaaactac gggtacggca tattatatgg tcgaagtatt aacgcaatta
     2641 ttcaagctgg tttaaatcca atggcgggtt ttctgcattc gtaccaaccg accaaaccgg
     2701 ttttggtgta tgacctggtg gaagagttcc gtcctctcgc cgttgatcgc ggtattttca
     2761 ccttgctgaa ccgtggtcag aagattgaac tgggcagaga ctctctttta agcgccaaaa
     2821 gtcggaagaa gatcgctaag tcagtaattg gatgcctgtc atcagaagtg tgctttaacg
     2881 gccaccgact gtcactcgag gacgtgatta aggaacaagc attaaatata aagaagcacc
     2941 tgtataacaa cgcaaagtat cgtccgttct taggcagatg gatgagtggc ggcctgaaaa
     3001 ttactcggag aatactgggc gatgctgaat tccatggaaa gcccgatcgg cttgaaaaga
     3061 gccggtctgt gtcaatcgga tctgtgttgt atgagtggat catagtcggc aggctcatcg
     3121 ctcaaacacc ttttcatttc ggcgatgaag agaaagccga gggcgcaatt ctcctcactc
     3181 cagataatcg gttcaggctg ccacgaacag cactgagggg aatcctgaga agggacttaa
     3241 aactcgccgg tgcatcagcc tgtgaggtgg aagtggggcg gagtgagcca tgcccttgtg
     3301 acgtgtgtaa gattatgcgc cgcgtgacac tgttggatac cgtgtctgag gaccttcggg
     3361 atttcctgcc agagttacgg aaaagaattc ggatcaatcc gcagagcggc actgtggcag
     3421 agggcgcgct gttcgacaca gaagtgggac ccgagggttt gtcctttccc ttcgtgctca
     3481 gatacaagtg cgagaaactg cccgacagct tgactacagt cctgtgttgg tggcaggaag
     3541 ggttggcttt cctttccgga gagagtgcga cagggaaagg tcggtttaga ttagagatta
     3601 acggcgcttt tgtatgggat ctcagcgacg agaatcaaag gaatgactat cttaagaatt
     3661 ggggatggcg tgacgagaag gggctcgagg aactgaagaa acgactgaac tcaggtctgc
     3721 cagagcccgg taattatagg gatccaaaat gggacttaat taagtaccag ctgaacattt
     3781 ccagccctct ccttcttaac gacccaatta gggctattgc tttgtacgag ggggaaggga
     3841 aagccccaga cgcagtgttc ttcaaaaagt acgtctttga aaatggaaag attgaggaaa
     3901 agccatgttt taaagctgag tccatcaggg gaatcttcag gaccgcagtg ggacggatca
     3961 agaatgtgtt aactaagaat cacgaggatt gcatttgtgt actgtgccac ctgttcggca
     4021 atgtgcatga aactgggaga ttaaagtttg aagatcttaa aatcgtctcc ggccaggagg
     4081 agaaattttt tgaccacgtg gcaattgacc gcttcttggg gggcgcaaaa gagaaataca
     4141 agttcgacga taaacccatc atcggggcac ccgacacccc tatcgtcctg gagggcaaga
     4201 tctgggtcaa gaaagatatc aatgatgaag caaaagagac actgagccag gccttctcag
     4261 atataaatac tggcatttac tacctggggg caaacggctc tatcggttac ggctggatcg
     4321 aagaggttaa ggctctcaag gatgataaga ggatttctcg cctcatgaat cctgcctttg
     4381 atgagacaga tgtagccgtt ccagaaaagc ccaaaactga tgccgaggtt cgcatcgagg
     4441 cagagaaagt atattaccca cactatttcg tcgaacccca taagaaggtg gaacgcgagg
     4501 agaaaccctg tggtcatcaa aagttccacg aggggcgact gactggtaaa attgagtgta
     4561 cactgaaaac gctgagccct ttaataatac cggacagtga ggtcgttacg gagaacggaa
     4621 acggccacaa gtcttaccag ttcttcagat taaataatga aatcatcatt ccgggaagcg
     4681 aaattcgggg tatgatcagt tccgtgtacg aagcacttac taactcttgc ttccgtgtcg
     4741 tggacgacat catcccggtg gtgatctccc gtagcgtgtc accgtaccag atcgctcaga
     4801 agctgcccag tccggatctg cgtccttgcg agcgtgagat tcttgaggac tcagaaattg
     4861 gatctttaga caagtgttca gagaaacgcc ttttccgcct gcatgaagac ggtctttgcc
     4921 catcgtgtcg tctgttcggc actacacatt ataagggacg cgttcggttc ggcttcgcaa
     4981 agcacgaggg cggcgaaaag tggctcatgg acgacaagac taaggacgag tatctgactt
     5041 taccgttgtt ggaacggccg cgcccaacat ggagcatgcc aagcgacgaa gacgacgtac
     5101 cgggacggaa attctacgta caccacaatg gttgggagaa agttaagcaa gatagtttag
     5161 ataataaaac gagcaaaacc gagaataatc gaagcgtgga accactcgat aaggataacc
     5221 tgtttaaatt cgacgtgttc ttcgagaacc tggaaccctg ggaacttggt ttgcttctct
     5281 actcattaga attggaagag gggttagccc acaagtttgg aatggcaaaa gcgttcggct
     5341 tcggctcgac aaagattgac gccgacaaaa tactgttacg tgaagaggcc gggaagttcc
     5401 acccttgcga gaaagcagac gaatacctta agaagggctt gaacaagctg accagctggt
     5461 tcggtaagaa cgaaatcaat gagcacatgc gaaacttact cttgttcatg acttactatc
     5521 agaaccttcc caaggttaag taccctgatt ttgacggata cgcaaaatgg cggtgcagct
     5581 acgttgaaca ggacaaggtg gaatactttc agaaccgctg gattgtggcc tcttaaaccg
     5641 gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta
     5701 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     5761 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     5821 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     5881 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     5941 gctcgatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     6001 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6061 aagcctagga tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6121 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggaa
     6181 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6241 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6301 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6361 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6421 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6481 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6541 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6601 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6661 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6721 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6781 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6841 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6901 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6961 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacact cagtggaacg
     7021 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7081 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7141 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7201 ccatagttgc ctgactcccc gtcgtgtag
//
