LOCUS       sequence-418030-        7358 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SynCas.v14A for analysis of RNA knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7358)
  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 7358)
  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.v14A mammalian expression
FEATURES             Location/Qualifiers
     source          1..7358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     542..561
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(780..884)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(963..982)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1154..1533
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1534..1737
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1688..1708
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5775..5792
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5815..5832)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5821..6045
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6116..6132)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6116..6132)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6129..6151)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6140..6156
                     /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(6164..6170,6171..6188,6189..6194))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6209..6230
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6347..6364)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6518..7106)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6598..6617)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7277..7358,1..710,711..779))
                     /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 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
       61 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
      121 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
      181 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
      241 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
      301 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
      361 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
      421 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
      481 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
      541 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
      601 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
      661 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
      721 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
      781 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
      841 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
      901 gtgccacctg acgtcgacgg atcgggagat cgatctcccg atcccctagg gtcgactctc
      961 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     1021 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     1081 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
     1141 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
     1201 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     1261 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     1321 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     1381 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     1441 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     1501 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     1561 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     1621 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     1681 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
     1741 ttagtgaacc gtcagatccg ctagagatcc gcggccgcat gacgactact atgaagattt
     1801 caattgaatt cctcgagccg tttcggatga ccaaatggca ggaaagcact aggcgaaaca
     1861 agaacaacaa ggaatttgtt cgcggccaag cttttgcaag atggcataga aataagaagg
     1921 ataacacaaa agggcggcct tacataactg ggacattgct gcggtccgca gtaattagaa
     1981 gcgcggaaaa cctgctgaca ttgagtgatg ggaaaattag cgaaaagaca tgttgtcctg
     2041 ggaaatttga tacggaagat aaagacaggt tgctgcaatt gaggcagcgg tccacccttc
     2101 gttggaccga taagaacccc tgtccagata acgctgaaac ttactgcccg ttttgcgaac
     2161 ttctcgggag atccggtaac gatggaaaga aggctgagaa gaaggattgg cgctttcgta
     2221 ttcattttgg caatctttcc cttccaggga agcccgattt cgacgggcca aaagctatag
     2281 gcagccaacg ggtacttaac agggtcgatt tcaaatccgg taaggcccat gacttcttta
     2341 aggcttacga agtcgaccat actaggtttc cccgcttcga aggggagatt accatagata
     2401 ataaagtcag cgctgaagcc aggaaactgc tctgtgattc tctcaagttt acggatcggc
     2461 tgtgtggagc tctgtgcgta atcaggttcg atgaatatac accagcagcc gatagtggga
     2521 aacaaaccga aaatgtccaa gcagaaccga atgctgcggt taagaaggct gcgggagatc
     2581 tggttaacag cgcgttaaac tacgggtacg gcatattata tggtcgaagt attaacgcaa
     2641 ttattcaagc tggtttaaat ccaatggcgg gttttctgca ttcgtaccaa ccgaccaaac
     2701 cggttttggt gtatgacctg gtggaagagt tccgtcctct cgccgttgat cgcggtattt
     2761 tcaccttgct gaaccgtggt cagaagattg aactgggcag agactctctt ttaagcgcca
     2821 aaagtcggaa gaagatcgct aagtcagtaa ttggatgcct gtcatcagaa gtgtgcttta
     2881 acggccaccg actgtcactc gaggacgtga ttaaggaaca agcattaaat ataaagaagc
     2941 acctgtataa caacgcaaag tatcgtccgt tcttaggcag atggatgagt ggcggcctga
     3001 aaattactcg gagaatactg ggcgatgctg aattccatgg aaagcccgat cggcttgaaa
     3061 agagccggtc tgtgtcaatc ggatctgtgt tgaaagaaac tgtggtatgc ggcgaactgg
     3121 tcgctaaaac gcccttcttc ttcggagcga tagacgaaga tgcaaaacaa accgacctgc
     3181 aagtactcct cactcccgat aacaagtata gactgccaag aagcgccgtg cgaggtatac
     3241 tccgtcgaga tcttcaaacc tattttgata gcccatgcaa tgctgagttg ggtggacggc
     3301 catgcatgtg taagacgtgt aggattatga gagggatcac ggtgatggat gcgcgcagtg
     3361 agtacaatgc cccgccagaa ataaggcatc gtacccgcat taatcccttc acaggcacgg
     3421 ttgcggaagg tgccctgttt aatatggaag tagcccccga ggggattgtc tttccatttc
     3481 aactccggta ccggggctct gaagatgggc tgcccgatgc actgaaaacg gtgttgaaat
     3541 ggtgggctga ggggcaggca ttcatgagtg gcgctgcctc aaccgggaag ggccgattcc
     3601 ggatggaaaa tgctaaatat gaaacgctgg atctgagcga cgagaatcaa aggaatgact
     3661 atcttaagaa ttggggatgg cgtgacgaga aggggctcga ggaactgaag aaacgactga
     3721 actcaggtct gccagagccc ggtaattata gggatccaaa atggcacgag attaacgttt
     3781 ccattgagat ggcaagccct tttattaatg gcgacccaat ccgcgcagcc gtggacaaac
     3841 gtggtacaga tgtggttacc ttcgttaagt ataaagctga aggggaagag gcgaaacccg
     3901 tatgtgcata caaggccgaa tcttttagag gggtgatcag aagtgccgtg gcacgcattc
     3961 atatggaaga tggcgtccct ttgactgagt tgactcacag tgactgtgaa tgtctcctgt
     4021 gccaaatctt tggaagtgag tatgaagccg gcaaaataag gtttgaagat ctcgtattcg
     4081 aaagtgaccc ggaacctgtg accttcgatc atgtggccat cgatagattc actggcggtg
     4141 cagctgataa gaagaaattc gatgattccc ctctgcccgg tagccctgca agaccgctca
     4201 tgttgaaagg ctccttctgg atccgcaggg acgttctcga ggacgaagag tactgtaagg
     4261 cactcggtaa ggctcttgca gatgtgaata atggccttta tcccctcggt ggaaagagcg
     4321 ccatcggcta cggacaggtc aagagtctgg gtataaaggg agatgataag aggatttctc
     4381 gcctcatgaa tcctgccttt gatgagacag atgtagccgt tccagaaaag cccaaaactg
     4441 atgccgaggt tcgcatcgag gcagagaaag tatattaccc acactatttc gtcgaacccc
     4501 ataagaaggt ggaacgcgag gagaaaccct gtggtcatca aaagttccac gaggggcgac
     4561 tgacaggtaa aattcggtgt aagctcatta ccaagacacc cctcatcgtc ccagatacta
     4621 gtaatgacga tttcttcaga cctgcggata aagaagctcg gaaggaaaag gacgaatatc
     4681 ataaatcata tgctttcttc agacttcata aacaaatcat gattcccggg agcgaattga
     4741 gaggaatggt gagtagtgtc tacgaaactg tgacaaattc ttgcttcagg atatttgatg
     4801 agactaaacg gttgtcatgg cggcgtagcg tgtcaccgta ccgaactgta gacgatagga
     4861 tgatcggcaa acgtatgagc gccgatctgc ggccgtgcca tggagattgg gtggaagatg
     4921 gtgatctcag tgccttgaat gcatatcccg agaaaagact cctcttgcgc caccccaaag
     4981 gactctgccc tgcttgccgg ctctttggaa ccggatctta caagggcaga gtcaggtttg
     5041 gattcgcgtc actcgaaaac gatccggagt ggctgatccc aggcaagaat cccggcgatc
     5101 cgtttcacgg cgggccggtg atgctctcat tgttggaacg gcctcgcccg acttggagta
     5161 taccgggatc cgacaataag tttaaagtgc ctggcagaaa gttttacgtc caccaccacg
     5221 cctggaaaac cattaaggac gggaaccatc ccacaacagg caaagctatt gaacaaagcc
     5281 ctaataaccg cactgtagaa gctctcgccg gcgggaattc ctttagcttc gaaattgcct
     5341 ttgagaacct gaaagaatgg gagctgggtt tgctcatcca cagcctgcaa ctcgaaaagg
     5401 gtctggcgca taaacttgga atggcaaagt ctatgggatt tggttcagtt gaaattgacg
     5461 tcgaatcagt gcgcctgaga aaagattgga agcaatggcg gaatggcaat tccgaaattc
     5521 ccaactggtt gggaaaagga tttgctaaac tgaaggaatg gttccgggac gagctcgatt
     5581 ttatagaaaa tcttaagaaa cttctttggt ttcctgaggg cgaccaagca ccccgggttt
     5641 gctaccccat gctgcgaaag aaggacgatc ctaatgggaa tagcggttac gaagaactca
     5701 aagacgggga attcaagaaa gaagatcggc agaagaagct gaccacgccg tggacaccgt
     5761 gggcataaac cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga
     5821 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     5881 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     5941 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     6001 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     6061 gaaccagctg gggctcgata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     6121 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6181 gcataaagtg taaagcctag gatgcctaat gagtgagcta actcacatta attgcgttgc
     6241 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6301 aacgcgcggg aagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6361 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6421 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6481 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6541 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6601 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6661 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6721 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6781 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6841 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6901 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6961 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7021 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7081 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgaca
     7141 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7201 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7261 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7321 tatttcgttc atccatagtt gcctgactcc ccgtcgtg
//
