LOCUS       sequence-478981-        9577 bp    DNA     circular SYN 17-JUL-2025
DEFINITION  Expression of array of 48 crRNAs targeting H3C1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9577)
  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 9577)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 17, 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-48xH3C1 array
FEATURES             Location/Qualifiers
     source          1..9577
                     /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        2216..2595
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          2876..3892
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3816..3838
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3900..3919
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    5967..6555
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6020..6040)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6438..6449)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6630..6863
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6982..7000)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7068..7172
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7173..7241,7242..8033)
                     /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(7391..7410)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8204..8792
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8693..8712
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8946..8963
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9203..9224
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9239..9244,9245..9262,9263..9269)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9277..9293
                     /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     9282..9304
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9301..9317
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9301..9317
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9367..9386
                     /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 gactaggaca
     2221 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     2281 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     2341 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     2401 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     2461 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     2521 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     2581 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc
     2641 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
     2701 gcgggggggg ggggggcgcg ccccaggggg gggggggggg ggcggggggg gggggggggg
     2761 gggggggcgg gggggggggg gggcgcccat ccaggccccg cccctcgcag gctgccctgc
     2821 tctggagcac cacgcgtcgc ggggccccta taaaaagcga agcgcgcggc gggcgggagt
     2881 cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg
     2941 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
     3001 ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct
     3061 tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg
     3121 tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg
     3181 gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg
     3241 tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt
     3301 gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc
     3361 tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc
     3421 ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc
     3481 gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg
     3541 tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg
     3601 acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc
     3661 tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
     3721 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg
     3781 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
     3841 cggctctagc gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
     3901 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gggcggccgc atcttgtgga
     3961 aaggacgaaa caccgaattt ctactaagtg tagatgatgc acccgttgcc ttagtaattt
     4021 ctactaagtg tagatcatac acctatctat atacaaattt ctactaagtg tagataagag
     4081 agcacccact gattcaattt ctactaagtg tagatatctg tgtactacag ctcttaattt
     4141 ctactaagtg tagatggtgt ctactgttga tgcataattt ctactaagtg tagatgattc
     4201 cagctttcca aagaaaattt ctactaagtg tagatagagc tgccaacgac acaccaattt
     4261 ctactaagtg tagatgcatt gccaagagct aatgcaattt ctactaagtg tagatcattg
     4321 accatggctt agaataattt ctactaagtg tagataatat gagatgacat ctcacaattt
     4381 ctactaagtg tagataataa cactcctaag tcaataattt ctactaagtg tagatactct
     4441 gaggtataca aacagaattt ctactaagtg tagatggatt caaagagcat aatcaaattt
     4501 ctactaagtg tagataaata agtccgaacc atgctaattt ctactaagtg tagatctact
     4561 caatctatca aagataattt ctactaagtg tagatcggag aggaggtgag cagtaaattt
     4621 ctactaagtg tagatgagct gcaacccaaa atcccaattt ctactaagtg tagatcagtt
     4681 accactgctg aacataattt ctactaagtg tagatcttag tgcagttcaa ttctaaattt
     4741 ctactaagtg tagatgaggt ccgttgccag agatgaattt ctactaagtg tagatcattg
     4801 cattggaatg acataaattt ctactaagtg tagatgtgta ctcaaattaa gcacaaattt
     4861 ctactaagtg tagataatta ataaccctaa tcaaaaattt ctactaagtg tagatataga
     4921 actagtattc ttctaaattt ctactaagtg tagataagga taaacctagg tagctaattt
     4981 ctactaagtg tagatactcc agaacattag gtttgaattt ctactaagtg tagattgagt
     5041 aatagtgtaa tcctaaattt ctactaagtg tagatagcta atataaacaa taccgaattt
     5101 ctactaagtg tagattaagc aaggaataag aataaaattt ctactaagtg tagatgagcg
     5161 atgtgtctcg cgaataattt ctactaagtg tagatagaga gaagcggtac cctctaattt
     5221 ctactaagtg tagatgaagg gggagtgata aatcaaattt ctactaagtg tagatgctgg
     5281 caagaaggca aagaaaattt ctactaagtg tagatacata gcctcatcta gtaagaattt
     5341 ctactaagtg tagatgagaa gcataacatc atgagaattt ctactaagtg tagatcgtgt
     5401 gtgcatatga gttaaaattt ctactaagtg tagataactg agaagcataa catataattt
     5461 ctactaagtg tagatctaaa gagctgaagg ttataaattt ctactaagtg tagatgtgct
     5521 gaacagaata ccagaaattt ctactaagtg tagatagata cctcgcttat tacacaattt
     5581 ctactaagtg tagatagaca tggtgactaa cacagaattt ctactaagtg tagatgccta
     5641 aattgtgttc ccccaaattt ctactaagtg tagatctaca ggcttcagtg gaagcaattt
     5701 ctactaagtg tagataatgc atgccttctc aataaaattt ctactaagtg tagatgggca
     5761 tgttgtatgg atgaaaattt ctactaagtg tagatccatg gtactccttg tccccaattt
     5821 ctactaagtg tagatatttc atcgcatgta aacctaattt ctactaagtg tagataggaa
     5881 aagcatactg tccataattt ctactaagtg tagataagtg gaaactcctg aatcagaatt
     5941 ctcgacctcg agaatcgata gatcctaatc aacctctgga ttacaaaatt tgtgaaagat
     6001 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     6061 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     6121 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     6181 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     6241 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     6301 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     6361 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     6421 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     6481 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     6541 gggccgcctc cccgcctgag atcctttaag accaatgact tacaaggcag ctgtagatct
     6601 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     6661 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     6721 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     6781 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     6841 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     6901 acttgcaaag aaatgaatat cagagagtga gaggcccggg ttaattaagg aaagggctag
     6961 atcattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     7021 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     7081 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     7141 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     7201 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     7261 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     7321 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     7381 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     7441 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     7501 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     7561 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     7621 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     7681 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     7741 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     7801 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     7861 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     7921 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     7981 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     8041 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     8101 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     8161 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     8221 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     8281 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     8341 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     8401 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     8461 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     8521 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     8581 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     8641 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     8701 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     8761 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     8821 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     8881 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     8941 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     9001 ccgcgcgttg gccgattcat taatgcagca agctcatggc tgactaattt tttttattta
     9061 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     9121 tggaggccta ggcttttgca aaaagctccc cgtggcacga caggtttccc gactggaaag
     9181 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     9241 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     9301 caggaaacag ctatgacatg attacgaatt tcacaaataa agcatttttt tcactgcatt
     9361 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa
     9421 ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt
     9481 acctgtggtt tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa
     9541 attgtatttg ttaaatatgt actacaaact tagtagt
//
