LOCUS       sequence-479454-        9594 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of array of 48 crRNAs targeting CCAT1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9594)
  AUTHORS   Yang LZ, Min YH, Liu YX, Gao BQ, Liu XQ, Huang Y, Wang H, Yang L, 
            Liu ZJ, Chen LL
  TITLE     CRISPR-array-mediated imaging of non-repetitive and multiplex 
            genomic loci in living cells.
  JOURNAL   Nat Methods. 2024 Sep;21(9):1646-1657. doi: 
            10.1038/s41592-024-02333-3. Epub 2024 Jul 4.
  PUBMED    38965442
REFERENCE   2  (bases 1 to 9594)
  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/s41592-024-02333-3"; journalName: "Nat Methods"; date: 
            "2024-09"; volume: "21"; issue: "9"; pages: "1646-1657"
COMMENT     Sequence Label: pCAG-48xCCAT1 array
FEATURES             Location/Qualifiers
     source          1..9594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2274..2577
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2579..2856
                     /label=chicken beta-actin promoter
     intron          2857..3873
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3797..3819
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3881..3900
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    5984..6572
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6037..6057)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6455..6466)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6647..6880
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6999..7017)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7085..7189
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7190..7258,7259..8050)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7408..7427)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8221..8809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8710..8729
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8963..8980
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9220..9241
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9256..9261,9262..9279,9280..9286)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9294..9310
                     /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)."
     primer_bind     9299..9321
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9318..9334
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9318..9334
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9384..9403
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagttat
     2221 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     2281 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     2341 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     2401 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     2461 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     2521 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
     2581 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
     2641 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggg
     2701 cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg
     2761 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
     2821 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
     2881 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
     2941 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
     3001 taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc
     3061 cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc
     3121 gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt
     3181 gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg
     3241 ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt
     3301 gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac
     3361 ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc
     3421 ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggagggc
     3481 tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc
     3541 agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc
     3601 tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag
     3661 cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc
     3721 cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg
     3781 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag cgcctctgct
     3841 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg
     3901 ctgtctcatc attttggcaa agggcggccg catcttgtgg aaaggacgaa acaccgtaat
     3961 ttctactaag tgtagatata gggaaacatg aagaaactaa tttctactaa gtgtagatgc
     4021 ctagtggaga aaacagatta atttctacta agtgtagatc tgtgagcaca ccaaagaaat
     4081 aatttctact aagtgtagat ctctaccatg cctactgtaa taatttctac taagtgtaga
     4141 tggataacgg acacagccat ataatttcta ctaagtgtag atggactata tcagaactat
     4201 gataatttct actaagtgta gatgtatgaa tgtatgacct tcttaatttc tactaagtgt
     4261 agataatagg gaaatctgta cacataattt ctactaagtg tagatggagg cttccacagt
     4321 agacctaatt tctactaagt gtagatgtga tgagttacca gatggttaat ttctactaag
     4381 tgtagatgtt tatatcctca gttacaataa tttctactaa gtgtagatgc tccatcactg
     4441 ctggcgacta atttctacta agtgtagata gagaaacatg attagttctt aatttctact
     4501 aagtgtagat ggtctcaaaa tggataataa taatttctac taagtgtaga tcttactgat
     4561 gttaccacaa ataatttcta ctaagtgtag atgagaagaa ggactaaaaa tctaatttct
     4621 actaagtgta gattcaccag tactacaact tagtaatttc tactaagtgt agatgattgg
     4681 aaagtccaca tgcctaattt ctactaagtg tagatccttg tgccaggcaa cattctaatt
     4741 tctactaagt gtagatccca cagccatatg gtccgataat ttctactaag tgtagatcca
     4801 tgcatggtag atcctaataa tttctactaa gtgtagatgg gctccctgga tacttaagta
     4861 atttctacta agtgtagatc atggtagatt ctctataatt aatttctact aagtgtagat
     4921 ctggacaggg tattgccaac taatttctac taagtgtaga tctgagcaca gttctctcat
     4981 gtaatttcta ctaagtgtag atgagctagg catattgtat tctaatttct actaagtgta
     5041 gatgggtatc cttcattatc aagtaatttc tactaagtgt agatagacct cagagtctat
     5101 ctgttaattt ctactaagtg tagatcctat gaaaatggca aaggttaatt tctactaagt
     5161 gtagattccc ttatcagaat gtaaaataat ttctactaag tgtagataca agaggaaatg
     5221 atgaggctaa tttctactaa gtgtagatag ggtggaagag ctagaacata atttctacta
     5281 agtgtagatg cacgagtcta ccattgctct aatttctact aagtgtagat ccagaaaaga
     5341 taaacaagtt taatttctac taagtgtaga tgaataaatg agtaccttcg gtaatttcta
     5401 ctaagtgtag atcttggcaa ccagaatgca tgtaatttct actaagtgta gatatggatg
     5461 gaagtacaca ttctaatttc tactaagtgt agattcagga cacagtgata acaataattt
     5521 ctactaagtg tagatcattc tgataaggga taaaataatt tctactaagt gtagatggct
     5581 gtctgagaaa ccaaaataat ttctactaag tgtagatgac tataaaagcc ataggcttaa
     5641 tttctactaa gtgtagatac tgcattcata aggattggta atttctacta agtgtagatg
     5701 aatgtgctga tgctagcatt aatttctact aagtgtagat cctgccttgc aaggacacat
     5761 taatttctac taagtgtaga taatacccca aggatggagg ttaatttcta ctaagtgtag
     5821 atacctaaga aacagcaaag attaatttct actaagtgta gatgatctgc tctgcctcac
     5881 agataatttc tactaagtgt agatgaggca agaatgcaca tatttaattt ctactaagtg
     5941 tagatttttt ttgaattctc gacctcgaga atcgatagat cctaatcaac ctctggatta
     6001 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     6061 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     6121 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     6181 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     6241 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     6301 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     6361 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     6421 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     6481 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     6541 gagtcggatc tccctttggg ccgcctcccc gcctgagatc ctttaagacc aatgacttac
     6601 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     6661 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
     6721 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     6781 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     6841 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt
     6901 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gcccgggtta
     6961 attaaggaaa gggctagatc attcttgaag acgaaagggc ctcgtgatac gcctattttt
     7021 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     7081 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     7141 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     7201 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     7261 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     7321 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     7381 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     7441 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     7501 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     7561 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     7621 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     7681 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     7741 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     7801 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     7861 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     7921 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     7981 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     8041 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     8101 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     8161 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     8221 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     8281 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     8341 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     8401 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     8461 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     8521 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     8581 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     8641 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     8701 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     8761 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     8821 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     8881 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     8941 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     9001 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagcaagc tcatggctga
     9061 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     9121 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctccccgt ggcacgacag
     9181 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     9241 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     9301 cggataacaa tttcacacag gaaacagcta tgacatgatt acgaatttca caaataaagc
     9361 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     9421 ctggatcaac tggataactc aagctaacca aaatcatccc aaacttccca ccccataccc
     9481 tattaccact gccaattacc tgtggtttca tttactctaa acctgtgatt cctctgaatt
     9541 attttcattt taaagaaatt gtatttgtta aatatgtact acaaacttag tagt
//
