LOCUS       sequence-337349-        8421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for encoding a human codon-optimized 
            dCas13X.1-REPAIRv2 driven by CMV promoter,mCherry driven by CMV 
            promoter and U6-driven crRNAs cloning site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8421)
  AUTHORS   Xu C, Zhou Y, Xiao Q, He B, Geng G, Wang Z, Cao B, Dong X, Bai W, 
            Wang Y, Wang X, Zhou D, Yuan T, Huo X, Lai J, Yang H
  TITLE     Programmable RNA editing with compact CRISPR-Cas13 systems from 
            uncultivated microbes.
  JOURNAL   Nat Methods. 2021 May;18(5):499-506. doi: 
            10.1038/s41592-021-01124-4. Epub 2021 May 3.
  PUBMED    33941935
REFERENCE   2  (bases 1 to 8421)
  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"; doi: 
            "10.1038/s41592-021-01124-4"; journalName: "Nat Methods"; date: 
            "2021-05"; volume: "18"; issue: "5"; pages: "499-506"
COMMENT     Sequence Label: 
            CMV-dCas13X.1-REPAIRv2-SV40pA_CMV-mCherry-BGHpA_U6-BbsI-DR
FEATURES             Location/Qualifiers
     source          1..8421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(104..172,173..964)
                     /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"
     primer_bind     complement(322..341)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1135..1723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1624..1643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1817..2065
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1817..1837
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1988..2007
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        2156..2535
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2536..2739
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     intron          2831..2963
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             2996..3016
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5339..5359
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6578..6604
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(6632..6649)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6638..6845
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        6914..7217
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7218..7421
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      7450..7459
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7456..8166
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     DDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(7601..7619)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    8189..8413
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        join(8420..8421,1..103)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
       61 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
      121 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
      181 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
      241 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
      301 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
      361 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
      421 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
      481 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
      541 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
      601 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
      661 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
      721 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
      781 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
      841 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
      901 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
      961 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     1021 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     1081 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     1141 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     1201 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     1261 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     1321 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     1381 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     1441 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     1501 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     1561 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     1621 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     1681 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     1741 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     1801 ccctgattct ctcgaggagg gcctatttcc catgattcct tcatatttgc atatacgata
     1861 caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca
     1921 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
     1981 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
     2041 tatcttgtgg aaaggacgaa acaccggggt cttcgatatt caagcgtcgg aagacctgct
     2101 ggagcagccc ccgatttgtg gggtgattac agcttttttt gtggacggtg atatcggcat
     2161 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     2221 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     2281 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     2341 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     2401 tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     2461 tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc
     2521 atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg atagcggttt
     2581 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     2641 caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc
     2701 ggtaggcgtg tacggtggga ggtctatata agcagaggtc gtttagtgaa ccgtcagatc
     2761 actagtagct ttattgcggt agtttatcac agttaaattg ctaacgcagt cagtgctcga
     2821 ctgatcacag gtaagtatca aggttacaag acaggtttaa ggaggccaat agaaactggg
     2881 cttgtcgaga cagagaagat tcttgcgttt ctgataggca cctattggtc ttactgacat
     2941 ccactttgcc tttctctcca caggggtacc gaagccgcta gtcgacgcca ccatgcccaa
     3001 gaagaagcgg aaggtggccc aggtgagcaa gcagacctcc aagaagaggg agctgagcat
     3061 cgacgagtac cagggcgccc ggaagtggtg cttcaccatt gccttcaaca aggccctggt
     3121 gaaccgggac aagaacgacg gcctgttcgt ggaaagcctg ctgagacacg agaagtacag
     3181 caagcacgac tggtacgacg aagatacccg ggccctgatc aagtgcagca cccaggccgc
     3241 caacgccaag gctgaagccc tggcgaacta cttcagtgct taccggcata gccctggctg
     3301 cctgaccttc accgccgagg acgaactgcg gaccatcatg gagagagcct atgagcgggc
     3361 catcttcgag tgcagaagaa gagagacaga ggtgatcatc gagtttccca gcctgttcga
     3421 gggcgaccgg atcaccaccg ccggcgtggt gtttttcgtg agctttttcg tggaaagaag
     3481 agtgctggat cggctgtatg gagccgtgtc cggcctgaag aagaatgagg gacagtacaa
     3541 gctgacccgg aaggccctga gcatgtactg cctgaaggac agcagattca ccaaggcctg
     3601 ggataagcgg gtgctgctgt tcagagacat cctggcccag ctgggaagaa tccccgccga
     3661 ggcctacgag tactaccacg gcgagcaggg tgataagaag agagctaacg acaatgaggg
     3721 cacaaatccc aagcggcaca aggacaagtt catcgaattt gcactgcact acctggaagc
     3781 ccagcacagc gagatctgct tcggcagacg ccacatcgtg cgggaagagg ccggcgccgg
     3841 cgatgagcac aagaagcacc ggaccaaggg aaaggtggtg gtggacttca gcaagaagga
     3901 cgaggaccag agctactata tctccaagaa caacgtgatc gtgcggatcg acaagaacgc
     3961 cggccctaga agctaccgga tgggcctgaa cgagctgaag tacctcgtgc tgctgagcct
     4021 gcaggggaag ggcgacgatg ccatcgccaa gctgtacaga tacagacagc acgtggagaa
     4081 catcctggat gtggtgaagg tgaccgataa ggataaccac gtgttcctgc cccgcttcgt
     4141 gctggagcag cacggcatcg gcagaaaggc cttcaagcag cggatcgatg gacgggtgaa
     4201 gcacgtgcgg ggcgtgtggg agaagaagaa ggccgccacc aatgaaatga ccctgcacga
     4261 gaaggccaga gacatcctgc agtacgtgaa cgaaaactgc acccggtcct tcaaccctgg
     4321 cgaatacaac agactgctgg tgtgcctggt gggcaaggac gtggagaact ttcaggccgg
     4381 cctgaagcgg ctgcagctgg ccgaaaggat cgatggccgg gtgtactcca tcttcgccca
     4441 gaccagcacc atcaatgaga tgcaccaggt ggtgtgcgac cagatcctga accggctgtg
     4501 cagaatcggc gaccagaagc tgtacgatta cgtgggactg ggcaagaagg acgaaatcga
     4561 ctacaagcag aaggtggcct ggttcaagga gcacatcagc atccggagag gattcctgag
     4621 aaagaagttc tggtacgata gcaagaaggg attcgcaaag ctggtggagg aacacctgga
     4681 gtccggcggc ggccagcgcg acgtgggcct ggacaagaag tactaccaca tcgacgccat
     4741 cggcagattc gagggcgcca accccgccct gtacgagacc ctggccagag atcggctgtg
     4801 cctcatgatg gcccagtact tcctgggcag cgtgagaaag gaactgggca acaagattgt
     4861 gtggagcaac gacagcatcg aactgcctgt ggaaggctct gtgggaaatg agaagagcat
     4921 cgtgttctcc gtgtctgact acggcaagct gtacgtgctg gacgatgccg aattcctggg
     4981 ccggatctgc gaatacttca tgccccacga aaagggcaag atccggtacc acacagtgta
     5041 cgaaaagggc tttagagcat acaacgacct gcagaagaag tgcgtggagg ccgtgctggc
     5101 tttcgaagag aaggtggtga aggccaagaa gatgagcgag aaggaaggcg cccactacat
     5161 cgacttccgg gagatcctgg cccagaccat gtgcaaggag gccgagaaaa ccgcagtgaa
     5221 caaggtggcg gctgccttct tcgctgcgca cctgaagttc gtgattgacg agttcggcct
     5281 gttcagcgac gtgatgaaga agtacggcat cgagaaggaa tggaagttcc ctgtcaagcc
     5341 caagaagaag cggaaggtgg gtggaggcgg aggttctggg ggaggaggta gtggcggtgg
     5401 tggttcagga ggcggcggaa gccagctgca tttaccgcag gttttagctg acgctgtctc
     5461 acgcctggtc ctgggtaagt ttggtgacct gaccgacaac ttctcctccc ctcacgctcg
     5521 cagaaaagtg ctggctggag tcgtcatgac aacaggcaca gatgttaaag atgccaaggt
     5581 gataagtgtt tctacaggag gcaaatgtat taatggtgaa tacatgagtg atcgtggcct
     5641 tgcattaaat gactgccatg cagaaataat atctcggaga tccttgctca gatttcttta
     5701 tacacaactt gagctttact taaataacaa agatgatcaa aaaagatcca tctttcagaa
     5761 atcagagcga ggggggttta ggctgaagga gaatgtccag tttcatctgt acatcagcac
     5821 ctctccctgt ggagatgcca gaatcttctc accacatgag ccaatcctgg aagaaccagc
     5881 agatagacac ccaaatcgta aagcaagagg acagctacgg accaaaatag agtctggtca
     5941 ggggacgatt ccagtgcgct ccaatgcgag catccaaacg tgggacgggg tgctgcaagg
     6001 ggagcggctg ctcaccatgt cctgcagtga caagattgca cgctggaacg tggtgggcat
     6061 ccagggatca ctgctcagca ttttcgtgga gcccatttac ttctcgagca tcatcctggg
     6121 cagcctttac cacggggacc acctttccag ggccatgtac cagcggatct ccaacataga
     6181 ggacctgcca cctctctaca ccctcaacaa gcctttgctc agtggcatca gcaatgcaga
     6241 agcacggcag ccagggaagg cccccaactt cagtgtcaac tggacggtag gcgactccgc
     6301 tattgaggtc atcaacgcca cgactgggaa ggatgagctg ggccgcgcgt cccgcctgtg
     6361 taagcacgcg ttgtactgtc gctggatgcg tgtgcacggc aaggttccct cccacttact
     6421 acgctccaag attaccaagc ccaacgtgta ccatgagtcc aagctggcgg caaaggagta
     6481 ccaggccgcc aaggcgcgtc tgttcacagc cttcatcaag gcggggctgg gggcctgggt
     6541 ggagaagccc accgagcagg accagttctc actcacgtac ccatacgacg taccagatta
     6601 cgcttaagaa ttcctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     6661 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     6721 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     6781 gggggtgggg tggggcagga cagcaagggg gaggattggg aagagaatag caggcatgct
     6841 ggggagcggc cgctagttat taatagtaat caattacggg gtcattagtt catagcccat
     6901 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     6961 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     7021 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     7081 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     7141 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     7201 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     7261 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     7321 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     7381 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     7441 tccgctagcg ccaccatggt gagcaagggc gaggaggata acatggccat catcaaggag
     7501 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag
     7561 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     7621 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
     7681 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc
     7741 ttcaagtggg agcgcgtgat gaacttcgat gacggcggcg tggtgaccgt gacccaggac
     7801 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc
     7861 tccgacggcc ccgtaatgca gaagaaaacc atgggctggg aggcctcctc cgagcggatg
     7921 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc
     7981 ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     8041 ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc
     8101 gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac
     8161 aagtaaaccg gtctcttggc ccggatccct gtgccttcta gttgccagcc atctgttgtt
     8221 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     8281 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     8341 gtggggcagg acagcaaggg ggaggattgg gaaaacaata gcaggcatgc tggggatgcg
     8401 gtgggctcta tggaagcttc g
//
