LOCUS       sequence-478948-        9659 bp    DNA     circular SYN 17-JUL-2025
DEFINITION  Expression of array of 48 crRNAs targeting GPBP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9659)
  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 9659)
  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-48xGPBP1 array
FEATURES             Location/Qualifiers
     source          1..9659
                     /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          2922..3938
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3862..3884
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3946..3965
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    6049..6637
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6102..6122)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6520..6531)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6712..6945
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7064..7082)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7150..7254
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7255..7323,7324..8115)
                     /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(7473..7492)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8286..8874
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8775..8794
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9028..9045
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9285..9306
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9321..9326,9327..9344,9345..9351)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9359..9375
                     /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     9364..9386
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9383..9399
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9383..9399
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9449..9468
                     /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 gggggggggg ggcccccggg gggggtgggg gggggggggg gggggggggg
     2761 gggggcgggc gcgggggggg gggggggggg gcggggggcg ggggggggcg aggcggagag
     2821 gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc
     2881 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc
     2941 ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg
     3001 cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct
     3061 tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg
     3121 agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga
     3181 gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc
     3241 tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg
     3301 gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag
     3361 ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg
     3421 agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc
     3481 cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg
     3541 agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga
     3601 gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc
     3661 aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg
     3721 cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc
     3781 gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc
     3841 gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagcgcct
     3901 ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta
     3961 ttgtgctgtc tcatcatttt ggcaaagggc ggccgcatct tgtggaaagg acgaaacacc
     4021 gtaatttcta ctaagtgtag atgccccggc ggcttggttg ggtaatttct actaagtgta
     4081 gatagaagcg ggtttgccct acttaatttc tactaagtgt agattcgggt gcttgcttgg
     4141 gtaataattt ctactaagtg tagatactct ggctacccat tttagtaatt tctactaagt
     4201 gtagatagca gttgtcttac tccttgtaat ttctactaag tgtagataga caaacggtag
     4261 ggtagcataa tttctactaa gtgtagataa tgtggtacag ccaagttgta atttctacta
     4321 agtgtagatt cgggagttgg ccactttaat aatttctact aagtgtagat cgttccaccc
     4381 gtttttcagg taatttctac taagtgtaga tacacctggg aagttggcat ttaatttcta
     4441 ctaagtgtag atcgaaaagg gccctgtacc tttaatttct actaagtgta gatcacttat
     4501 ctctgagtta cattaatttc tactaagtgt agatcaggtg attgaattac tcagtaattt
     4561 ctactaagtg tagatagttt cagaactaga gtagttaatt tctactaagt gtagatgctt
     4621 gtctgtatcc ctatattaat ttctactaag tgtagataac ctacagcaag aatgtaataa
     4681 tttctactaa gtgtagatta gaggcagcgt tttgaaagta atttctacta agtgtagatt
     4741 ccttgtgctt ctagttgtgt aatttctact aagtgtagat tctctcaggt ggtaggaatt
     4801 taatttctac taagtgtaga taactcaagt ctattagggg ctaatttcta ctaagtgtag
     4861 atatgcttgt atcaccctct ggtaatttct actaagtgta gatagcaatt aataatgccc
     4921 ctctaatttc tactaagtgt agatccgtcc ccaaccttct gttttaattt ctactaagtg
     4981 tagatacaag atgcagggaa tcaactaatt tctactaagt gtagattatt aactgctgga
     5041 gaattttaat ttctactaag tgtagatata gactcagatt aatctggtaa tttctactaa
     5101 gtgtagatta ttcgattacc atccttagta atttctacta agtgtagatc ataggatcag
     5161 gaaggagcat aatttctact aagtgtagat ccagactcag cttttaaaac taatttctac
     5221 taagtgtaga ttgaaagcca aactgtcttc ctaatttcta ctaagtgtag atacgttaca
     5281 gaatatggaa gttaatttct actaagtgta gattggtagt tgtacatgca ttataatttc
     5341 tactaagtgt agataaaatc tcgaatgggc tctgtaattt ctactaagtg tagatatttt
     5401 gaagtacagt aggactaatt tctactaagt gtagataaaa gtgaagggcg gaccagtaat
     5461 ttctactaag tgtagatatg gaatgtttct agaagattaa tttctactaa gtgtagatga
     5521 gggaaggtgg ggagaggtta atttctacta agtgtagatg gaaaggaaat ccaccatatt
     5581 aatttctact aagtgtagat ttaaaagatg gtcacatgaa taatttctac taagtgtaga
     5641 ttgtgtaagc atgatatcaa ataatttcta ctaagtgtag ataaaatcag tattaccgtg
     5701 tttaatttct actaagtgta gatgtgacac taggacatgt ttttaatttc tactaagtgt
     5761 agatattttg ctggtaattg tgattaattt ctactaagtg tagatttcaa gaagtataag
     5821 aaggttaatt tctactaagt gtagatgctc aaaatcaaaa ctcagttaat ttctactaag
     5881 tgtagattcc tcttaggttt ctttccataa tttctactaa gtgtagattt attcatacct
     5941 gctaagagta atttctacta agtgtagatt agtgtcttct gtcacatgat aatttctact
     6001 aagtgtagat tgaagctgtt cattgataac cgagaatcga tagatcctaa tcaacctctg
     6061 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     6121 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     6181 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     6241 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     6301 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     6361 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     6421 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     6481 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     6541 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     6601 cagacgagtc ggatctccct ttgggccgcc tccccgcctg agatccttta agaccaatga
     6661 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     6721 taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag
     6781 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     6841 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     6901 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     6961 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcccg
     7021 ggttaattaa ggaaagggct agatcattct tgaagacgaa agggcctcgt gatacgccta
     7081 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     7141 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     7201 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     7261 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     7321 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     7381 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     7441 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     7501 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     7561 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     7621 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     7681 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     7741 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     7801 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     7861 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     7921 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     7981 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     8041 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     8101 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     8161 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     8221 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     8281 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     8341 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     8401 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     8461 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     8521 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     8581 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     8641 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     8701 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     8761 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     8821 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     8881 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     8941 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     9001 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     9061 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag caagctcatg
     9121 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     9181 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc cccgtggcac
     9241 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     9301 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     9361 gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa tttcacaaat
     9421 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     9481 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca
     9541 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct
     9601 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagt
//
