LOCUS       sequence-418033-        7169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SynCas.v10 for analysis of RNA knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7169)
  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 7169)
  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.v10 mammalian expression
FEATURES             Location/Qualifiers
     source          1..7169
                     /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             5583..5600
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5623..5640)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5629..5853
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5924..5940)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5924..5940)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5937..5959)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5948..5964
                     /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(5972..5978,5979..5996,5997..6002))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6017..6038
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6155..6172)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6326..6914)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6406..6425)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7085..7169,1..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 ctaatctcgc tgaaaagacc gcagagcaaa
     2581 ttattagcat tctggacgat aataagaaaa ccgaatatac ccgcctgctt gctgatgcta
     2641 ttcgttcact ccgccgctct agtaaacttg tcgctggctt gccaaaggat catgacggga
     2701 aggatgacca ttacctctgg gatatcggca agaagaagaa agacgaaaac tctgtcacta
     2761 ttaggcaaat ccttacgacc tcagcagaca ccaaggaact caagaatgcc ggaaaatgga
     2821 gagaattctg cgagaagctg ggtgaagcgc tgtacctcaa gagtaaagac atgagtggcg
     2881 gcctgaaaat tactcggaga atactgggcg atgctgaatt ccatggaaag cccgatcggc
     2941 ttgaaaagag ccggtctgtg tcaatcggat ctgtgttgaa agaaactgtg gtatgcggcg
     3001 aactggtcgc taaaacgccc ttcttcttcg gagcgataga cgaagatgca aaacaaaccg
     3061 acctgcaagt actcctcact cccgataaca agtatagact gccaagaagc gccgtgcgag
     3121 gtatactccg tcgagatctt caaacctatt ttgatagccc atgcaatgct gagttgggtg
     3181 gacggccatg catgtgtaag acgtgtagga ttatgagagg gatcacggtg atggatgcgc
     3241 gcagtgagta caatgccccg ccagaaataa ggcatcgtac ccgcattaat cccttcacag
     3301 gcacggttgc ggaaggtgcc ctgtttaata tggaagtagc ccccgagggg attgtctttc
     3361 catttcaact ccggtaccgg ggctctgaag atgggctgcc cgatgcactg aaaacggtgt
     3421 tgaaatggtg ggctgagggg caggcattca tgagtggcgc tgcctcaacc gggaagggcc
     3481 gattccggat ggaaaatgct aaatatgaaa cgctggatct gagcgacgag aatcaaagga
     3541 atgactatct taagaattgg ggatggcgtg acgagaaggg gctcgaggaa ctgaagaaac
     3601 gactgaactc aggtctgcca gagcccggta attataggga tccaaaatgg acaaaagtag
     3661 ggtatgagat taaaatacag tcgccccttc ttagcaacga tcctatcgca gcattgttcg
     3721 acactgcaaa ccacgacgcg gtcgcgtata agaaacgagt gtgggagaac ggacgtatca
     3781 tagagcgaca cactttcaaa ggagagtcta tacgaggggt gctgcgcact gcacttgggc
     3841 gtgagtacgg attgttcgaa ttaaaacacg aagactgtcc ttgtagtctt tgcacaattt
     3901 tcggtaacga acacgaagcg ggcaaaatac gtttcgaaga cttagaaatc gaggacggtg
     3961 ttgccgaaaa gcacttcgat cacgttgcta tcgatcgctt cacaggcgga gccgtagaca
     4021 agaagaaatt cgacgatttc tctttagctg gttccccggg aaagcccatt aaactcaaag
     4081 ggtgcttttg gttgcgtgcg gatatattta gtgacgagaa gaaggacgag aagaagaaaa
     4141 taataaacgc tctttgtgac gtgcgtgatg gcttgtaccc tctcggtggc aagggcggtg
     4201 tcggatacgg gtgggtaaaa gagctgagaa taacggacgg cccagacgat tttaagctcc
     4261 cggaaccgaa atgtaaggaa attccgccag acggtactcc aagcgattac ccttttccgc
     4321 agttaaagcc actgcaaaca gagaaggata aagtttactc gccccactac tttctgaaac
     4381 cagcagagaa gaaagttgac cggaactggg aaaagcggct gatcggacac gagcgtttcg
     4441 ataaggacct gttgactggt aaaattgagt gtacactgaa aacgctgagc cctttaataa
     4501 taccggacag tgaggtcgtt acggagaacg gaaacggcca caagtcttac cagttcttca
     4561 gattaaataa tgaaatcatc attccgggaa gcgaaattcg gggtatgatc agttccgtgt
     4621 acgaagcact tactaactct tgcttccgtg tcgtggacga catcatcccg gtggtgatct
     4681 cccgtagcgt gtcaccgtac cagatcgctc agaagctgcc cagtccggat ctgcgtcctt
     4741 gcgagcgtga gattcttgag gactcagaaa ttggatcttt agacaagtgt tcagagaaac
     4801 gccttttccg cctgcatgaa gacggtcttt gcccatcgtg tcgtctgttc ggcactacac
     4861 attataaggg acgcgttcgg ttcggcttcg caaagcacga gggcggcgaa aagtggctca
     4921 tggacgacaa gactaaggac gagtatctga ctttaccgtt gttggaacgg ccgcgcccaa
     4981 catggagcat gccaagcgac gaagacgacg taccgggacg gaaattctac gtacaccaca
     5041 atggttggga gaaagttaag caagatagtt tagataataa aacgagcaaa accgagaata
     5101 atcgaagcgt ggaaccactc gataaggata acctgtttaa attcgacgtg ttcttcgaga
     5161 acctggaacc ctgggaactt ggtttgcttc tctactcatt agaattggaa gaggggttag
     5221 cccacaagtt tggaatggca aaagcgttcg gcttcggctc gacaaagatt gacgccgaca
     5281 aaatactgtt acgtgaggct ccgaacgagt ggcaagatgc taccaatcgt aaggacgaca
     5341 tgttacgcaa gggcttcggc gagctgacat cctggttcga caaggattgg gacaaagtcg
     5401 agcacataga cggtctgcgt tccttacttc agattccgga agcgctttca agagccgaag
     5461 tgcgctaccc ggaccttacg gaagacgacc cagagaagcc cccgggctac gtgcaattgt
     5521 ctgataagaa agagggccgc aagtacttga tccgtccctg gcaaccttgg tactaaaccg
     5581 gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta
     5641 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     5701 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     5761 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     5821 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     5881 gctcgatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5941 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6001 aagcctagga tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6061 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggaa
     6121 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6181 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6241 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6301 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6361 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6421 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6481 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6541 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6601 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6661 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6721 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6781 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6841 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6901 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacact cagtggaacg
     6961 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7021 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7081 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7141 ccatagttgc ctgactcccc gtcgtgtag
//
