LOCUS       sequence-418031-        7094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SynCas.v41 for analysis of RNA knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7094)
  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 7094)
  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.v41 mammalian expression
FEATURES             Location/Qualifiers
     source          1..7094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     535..554
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(773..877)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(956..975)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1147..1526
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1527..1730
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1681..1701
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5504..5521
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5544..5561)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5550..5774
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5845..5861)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5845..5861)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5858..5880)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5869..5885
                     /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(5893..5899,5900..5917,5918..5923))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5938..5959
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6076..6093)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6247..6835)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6327..6346)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7006..7094,1..703,704..772))
                     /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 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
       61 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
      121 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
      181 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
      241 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
      301 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
      361 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
      421 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
      481 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
      541 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
      601 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
      661 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
      721 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
      781 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
      841 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      901 ctgacgtcga cggatcggga gatcgatctc ccgatcccct agggtcgact ctcagtacaa
      961 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     1021 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     1081 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
     1141 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     1201 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     1261 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     1321 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     1381 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     1441 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     1501 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     1561 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     1621 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     1681 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     1741 accgtcagat ccgctagaga tccgcggccg catgacgact actatgaaga tttcaattga
     1801 attcctcgag ccgtttcgga tgaccaaatg gcaggaaagc actaggcgaa acaagaacaa
     1861 caaggaattt gttcgcggcc aagcttttgc aagatggcat agaaataaga aggataacac
     1921 aaaagggcgg ccttacataa ctgggacatt gctgcggtcc gcagtaatta gaagcgcgga
     1981 aaacctgctg acattgagtg atgggaaaat tagcgaaaag acatgttgtc ctgggaaatt
     2041 tgatacggaa gataaagaca ggttgctgca attgaggcag cggtccaccc ttcgttggac
     2101 cgataagaac ccctgtccag ataacgctga aacttactgc ccgttttgcg aacttctcgg
     2161 gagatccggt aacgatggaa agaaggctga gaagaaggat tggcgctttc gtattcattt
     2221 tggcaatctt tcccttccag ggaagcccga tttcgacggg ccaaaagcta taggcagcca
     2281 acgggtactt aacagggtcg atttcaaatc cggtaaggcc catgacttct ttaaggctta
     2341 cgaagtcgac catactaggt ttccccgctt cgaaggggag attaccatag ataataaagt
     2401 cagcgctgaa gccaggaaac tgctctgtga ttctctcaag tttacggatc ggctgtgtgg
     2461 agctctgtgc gtaatcaggt tcgatgaata tacaccagca gccgatagtg ggaaacaaac
     2521 cgaaaatgtc caagcagaac cgaatgctca gcttagtaag caggccaaag aagatattga
     2581 aaagatattt caggtggtta aagacgctca gaagctcaga ctgctgagtg actgttttag
     2641 agagttaaca agaatggcaa acaaagatga gctggctttg ccattaggcc cagaggacga
     2701 cggacattat ctgtgggata agattaaagt tgagggaaag acactgcgca tattcttgcg
     2761 gaattgtttt agtcaatata aggacaattg gttatgtttt tgcgacgaag ccagtaaaaa
     2821 gggctaccag aagtacagag agatgagtgg cggcctgaaa attactcgga gaatactggg
     2881 cgatgctgaa ttccatggaa agcccgatcg gcttgaaaag agccggtctg tgtcaatcgg
     2941 atctgtgttg tatgagtgga tcatagtcgg caggctcatc gctcaaacac cttttcattt
     3001 cggcgatgaa gagaaagccg agggcgcaat tctcctcact ccagataatc ggttcaggct
     3061 gccacgaaca gcactgaggg gaatcctgag aagggactta aaactcgccg gtgcatcagc
     3121 ctgtgaggtg gaagtggggc ggagtgagcc atgcccttgt gacgtgtgta agattatgcg
     3181 ccgcgtgaca ctgttggata ccgtgtctga ggaccttcgg gatttcctgc cagagttacg
     3241 gaaaagaatt cggatcaatc cgcagagcgg cactgtggca gagggcgcgc tgttcgacac
     3301 agaagtggga cccgagggtt tgtcctttcc cttcgtgctc agatacaagt gcgagaaact
     3361 gcccgacagc ttgactacag tcctgtgttg gtggcaggaa gggttggctt tcctttccgg
     3421 agagagtgcg acagggaaag gtcggtttag attagagatt aacggcgctt ttgtatggga
     3481 tctcagcgac gagaatcaaa ggaatgacta tcttaagaat tggggatggc gtgacgagaa
     3541 ggggctcgag gaactgaaga aacgactgaa ctcaggtctg ccagagcccg gtaattatag
     3601 ggatccaaaa tgggacttaa ttaagtacca gctgaacatt tccagccctc tccttcttaa
     3661 cgacccaatt agggctattg ctttgtacga gggggaaggg aaagccccag acgcagtgtt
     3721 cttcaaaaag tacgtctttg aaaatggaaa gattgaggaa aagccatgtt ttaaagctga
     3781 gtccatcagg ggaatcttca ggaccgcagt gggacggatc aagaatgtgt taactaagaa
     3841 tcacgaggat tgcatttgtg tactgtgcca cctgttcggc aatgtgcatg aaactgggag
     3901 attaaagttt gaagatctta aaatcgtctc cggccaggag gagaaatttt ttgaccacgt
     3961 ggcaattgac cgcttcttgg ggggcgcaaa agagaaatac aagttcgacg ataaacccat
     4021 catcggggca cccgacaccc ctatcgtcct ggagggcaag atctgggtca agaaagatat
     4081 caatgatgaa gcaaaagaga cactgagcca ggccttctca gatataaata ctggcattta
     4141 ctacctgggg gcaaacggct ctatcggtta cggctggatc gaagaggtta aggctctcaa
     4201 ggatgataag aggatttctc gcctcatgaa tcctgccttt gatgagacag atgtagccgt
     4261 tccagaaaag cccaaaactg atgccgaggt tcgcatcgag gcagagaaag tatattaccc
     4321 acactatttc gtcgaacccc ataagaaggt ggaacgcgag gagaaaccct gtggtcatca
     4381 aaagttccac gaggggcgac tgactggtaa aattgagtgt acactgaaaa cgctgagccc
     4441 tttaataata ccggacagtg aggtcgttac ggagaacgga aacggccaca agtcttacca
     4501 gttcttcaga ttaaataatg aaatcatcat tccgggaagc gaaattcggg gtatgatcag
     4561 ttccgtgtac gaagcactta ctaactcttg cttccgtgtc gtggacgaca tcatcccggt
     4621 ggtgatctcc cgtagcgtgt caccgtacca gatcgctcag aagctgccca gtccggatct
     4681 gcgtccttgc gagcgtgaga ttcttgagga ctcagaaatt ggatctttag acaagtgttc
     4741 agagaaacgc cttttccgcc tgcatgaaga cggtctttgc ccatcgtgtc gtctgttcgg
     4801 cactacacat tataagggac gcgttcggtt cggcttcgca aagcacgagg gcggcgaaaa
     4861 gtggctcatg gacgacaaga ctaaggacga gtatctgact ttaccgttgt tggaacggcc
     4921 gcgcccaaca tggagcatgc caagcgacga agacgacgta ccgggacgga aattctacgt
     4981 acaccacaat ggttgggaga aagttaagca agatagttta gataataaaa cgagcaaaac
     5041 cgagaataat cgaagcgtgg aaccactcga taaggataac ctgtttaaat tcgacgtgtt
     5101 cttcgagaac ctggaaccct gggaacttgg tttgcttctc tactcattag aattggaaga
     5161 ggggttagcc cacaagtttg gaatggcaaa agcgttcggc ttcggctcga caaagattga
     5221 cgccgacaaa atactgttac gtgaagaggc cgggaagttc cacccttgcg agaaagcaga
     5281 cgaatacctt aagaagggct tgaacaagct gaccagctgg ttcggtaaga acgaaatcaa
     5341 tgagcacatg cgaaacttac tcttgttcat gacttactat cagaaccttc ccaaggttaa
     5401 gtaccctgat tttgacggat acgcaaaatg gcggtgcagc tacgttgaac aggacaaggt
     5461 ggaatacttt cagaaccgct ggattgtggc ctcttaaacc ggtcatcatc accatcacca
     5521 ttgagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     5581 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     5641 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     5701 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     5761 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctcgatac cgtcgacctc
     5821 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5881 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctagg atgcctaatg
     5941 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6001 gtcgtgccag ctgcattaat gaatcggcca acgcgcggga agaggcggtt tgcgtattgg
     6061 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6121 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6181 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6241 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6301 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6361 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6421 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6481 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6541 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6601 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6661 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6721 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6781 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6841 tcctttgatc ttttctacgg ggtctgacac tcagtggaac gaaaactcac gttaagggat
     6901 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6961 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7021 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7081 cgtcgtgtag ataa
//
