LOCUS       sequence-478955-        9653 bp    DNA     circular SYN 17-JUL-2025
DEFINITION  Expression of array of 48 crRNAs targeting NFIL3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9653)
  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 9653)
  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-48xNFIL3 array
FEATURES             Location/Qualifiers
     source          1..9653
                     /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"
     primer_bind     3815..3837
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3899..3918
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    6043..6631
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6096..6116)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6514..6525)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6706..6939
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7058..7076)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7144..7248
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7249..7317,7318..8109)
                     /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(7467..7486)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8280..8868
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8769..8788
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9022..9039
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9279..9300
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9315..9320,9321..9338,9339..9345)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9353..9369
                     /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     9358..9380
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9377..9393
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9377..9393
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9443..9462
                     /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 gccccggggg gggggggggg gggggggggg gggggggggg
     2761 gggggggggg gggggggggg gggggcggca gggggggggg ggggggcaag tttttttttt
     2821 tctggggggg ggggggggcg gcggcccaat aaaaaaagag tggcgccggg ggggggggtc
     2881 gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg
     2941 ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc
     3001 tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt
     3061 gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt
     3121 gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg
     3181 cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt
     3241 gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg
     3301 ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct
     3361 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg
     3421 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
     3481 cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt
     3541 cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga
     3601 cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct
     3661 agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     3721 cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg
     3781 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     3841 ggctctagcg cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     3901 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag ggcggccgca tcttgtggaa
     3961 aggacgaaac accgtaattt ctactaagtg tagatgccag ttgagttgag ccgattaatt
     4021 tctactaagt gtagattgtg gccggtggac gtttggtaat ttctactaag tgtagatgga
     4081 accaagcgcg tctccattaa tttctactaa gtgtagattt ctggcgcggg gggtcctcta
     4141 atttctacta agtgtagatc gagctcgggt gggagtccct aatttctact aagtgtagat
     4201 gcagcgcccg gtgatttgct taatttctac taagtgtaga tactccacgc cttcggagac
     4261 ctaatttcta ctaagtgtag atcaagccct cgggaaggtt gttaatttct actaagtgta
     4321 gattgagagg cctttcggac gtttaatttc tactaagtgt agatattgca gcgccttcgc
     4381 gccgtaattt ctactaagtg tagattgtaa tgaatgcccc gcgcgtaatt tctactaagt
     4441 gtagattttc tcctcgccgg cccgagtaat ttctactaag tgtagatatc gctgcctccg
     4501 ccgcgcataa tttctactaa gtgtagatca gccgtcatct tcgccctata atttctacta
     4561 agtgtagatc acctcaacaa agtgtattct aatttctact aagtgtagat ggagagaaga
     4621 ctaccgtatc taatttctac taagtgtaga tccgatacac ggatagtggt ctaatttcta
     4681 ctaagtgtag atgaaagcct tccagggcag cgtaatttct actaagtgta gattgctcca
     4741 tttactcgag ttataatttc tactaagtgt agatcagtta ctccggccta gtgataattt
     4801 ctactaagtg tagatgccac tgagtccccc gtctctaatt tctactaagt gtagattggg
     4861 aatccatagc ataccgtaat ttctactaag tgtagatcaa tgaacacgat tagctgctaa
     4921 tttctactaa gtgtagatcg tctcttccgt agcccgggta atttctacta agtgtagatg
     4981 tgacctcctg tctggtctgt aatttctact aagtgtagat cctcctgtta caggcagtag
     5041 taatttctac taagtgtaga ttctggatac cctggtttta ctaatttcta ctaagtgtag
     5101 atacaattgt ggcctcgcag tgtaatttct actaagtgta gatctcatgc tgccgaatgt
     5161 gtgtaatttc tactaagtgt agatgtaatt gaggcgcagc gttgtaattt ctactaagtg
     5221 tagatcaccc tgtttgtcgt cagcgtaatt tctactaagt gtagatccag tcgtgttacg
     5281 caacaataat ttctactaag tgtagatgtg ctggggcttg agacgactaa tttctactaa
     5341 gtgtagatgg caagcctgtg agtcactgta atttctacta agtgtagata acgcactctt
     5401 gttcgtgatt aatttctact aagtgtagat aagacaactc gaccttcaag taatttctac
     5461 taagtgtaga tgtttaccct tacacttgcg ataatttcta ctaagtgtag attatccagg
     5521 ttggacttgt attaatttct actaagtgta gatggacgta atatagacgt aaataatttc
     5581 tactaagtgt agataccgtg ggcagattat gcaataattt ctactaagtg tagatgatag
     5641 agaaatcatc caagctaatt tctactaagt gtagatgcat cataccctac accgcttaat
     5701 ttctactaag tgtagatcag taggacagca gcgtctataa tttctactaa gtgtagataa
     5761 ctctggagat cagtgttcta atttctacta agtgtagata tcactatcaa tactggagct
     5821 aatttctact aagtgtagat tgaacctcac cgcagaacag taatttctac taagtgtaga
     5881 tcccccagac cagatatgac ctaatttcta ctaagtgtag atccctaaat gaaacgcacg
     5941 tttaatttct actaagtgta gatagacacg acaagacaaa tcataatttc tactaagtgt
     6001 agatctgcct tgtcaggatg ggaccgagaa tcgatagatc ctaatcaacc tctggattac
     6061 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     6121 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     6181 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     6241 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     6301 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     6361 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     6421 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     6481 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     6541 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     6601 agtcggatct ccctttgggc cgcctccccg cctgagatcc tttaagacca atgacttaca
     6661 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     6721 actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga
     6781 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     6841 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     6901 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta
     6961 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg cccgggttaa
     7021 ttaaggaaag ggctagatca ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     7081 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     7141 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     7201 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     7261 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     7321 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     7381 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     7441 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     7501 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     7561 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     7621 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     7681 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     7741 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     7801 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     7861 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     7921 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     7981 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     8041 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     8101 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     8161 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     8221 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     8281 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     8341 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     8401 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     8461 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     8521 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     8581 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     8641 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     8701 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     8761 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     8821 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     8881 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     8941 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     9001 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     9061 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagcaagct catggctgac
     9121 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     9181 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctccccgtg gcacgacagg
     9241 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     9301 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     9361 ggataacaat ttcacacagg aaacagctat gacatgatta cgaatttcac aaataaagca
     9421 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     9481 tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac cccataccct
     9541 attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta
     9601 ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt agt
//
