LOCUS       sequence-478950-        9063 bp    DNA     circular SYN 17-JUL-2025
DEFINITION  Expression of array of 36 crRNAs targeting MYC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9063)
  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 9063)
  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-36XMYC array
FEATURES             Location/Qualifiers
     source          1..9063
                     /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"
     promoter        2597..2874
                     /label=chicken beta-actin promoter
     intron          2875..3891
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     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    5453..6041
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5506..5526)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5924..5935)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6116..6349
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6468..6486)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6554..6658
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6659..6727,6728..7519)
                     /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(6877..6896)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7690..8278
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8179..8198
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8432..8449
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8689..8710
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8725..8730,8731..8748,8749..8755)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8763..8779
                     /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     8768..8790
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8787..8803
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8787..8803
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8853..8872
                     /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 ggcgccgggg ggggcggggg ggggggaggg ggggggcggg
     2761 gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
     2821 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
     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 ggcggccgct aatttctact
     3961 aagtgtagat ccccgacccc aacaccttct taatttctac taagtgtaga tgctacacaa
     4021 gaatctagaa ttaatttcta ctaagtgtag atgccggaga ccctggattc tataatttct
     4081 actaagtgta gatactagag gaccaactca gtataatttc tactaagtgt agatgcaggg
     4141 gctgataaca ttcttaattt ctactaagtg tagatgcagg aaatgtaaac gaagataatt
     4201 tctactaagt gtagatagta gcagccttgt ctagcctaat ttctactaag tgtagatcat
     4261 gagagaaaac aaagccataa tttctactaa gtgtagatac ataggaaggc acaaataata
     4321 atttctacta agtgtagatg ctgccttcca ggcattaatt aatttctact aagtgtagat
     4381 acttaaaaca gctcaataca taatttctac taagtgtaga tcaatgccca gatccactca
     4441 ctaatttcta ctaagtgtag atatggaact cactatgaaa cataatttct actaagtgta
     4501 gatcctggca cgtgtccctg gtctaatttc tactaagtgt agatgcttca attagagtaa
     4561 tgcctaattt ctactaagtg tagataaccc agcaaatgtc tattctaatt tctactaagt
     4621 gtagatagat aatctctttg tgattctaat ttctactaag tgtagataat gaagaggaaa
     4681 attaatctaa tttctactaa gtgtagatga gatggcacgt caccacatta atttctacta
     4741 agtgtagatc cagtctccac cactccctat aatttctact aagtgtagat gtctatgtac
     4801 ttgtgaatta taatttctac taagtgtaga ttgtgttgct caggctggtc ttaatttcta
     4861 ctaagtgtag atgtagagac agggtttcac cataatttct actaagtgta gatctttaat
     4921 gaatgcagta tactaatttc tactaagtgt agatttggag gtggtggagg gagataattt
     4981 ctactaagtg tagattctga agtgcagagt tggtataatt tctactaagt gtagatattc
     5041 tgtaagatga tcatggtaat ttctactaag tgtagatctc tagctacaca ctattcttaa
     5101 tttctactaa gtgtagatca tcgttggcct ctacccacta atttctacta agtgtagata
     5161 gacaggatct cactctgtct aatttctact aagtgtagat ttgcatactc attttagtat
     5221 taatttctac taagtgtaga tgtggagacg gggtttcacc ataatttcta ctaagtgtag
     5281 attacagggt gagaccctgt cttaatttct actaagtgta gatgtctggg gaactgcctg
     5341 aactaatttc tactaagtgt agatatgccg tggaataaca caaataattt ctactaagtg
     5401 tagatcagtg agacctcatc tctacttttt ttctcgagaa tcgatagatc ctaatcaacc
     5461 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     5521 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     5581 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     5641 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     5701 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     5761 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     5821 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     5881 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     5941 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     6001 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctgagatcc tttaagacca
     6061 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     6121 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
     6181 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     6241 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     6301 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
     6361 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     6421 cccgggttaa ttaaggaaag ggctagatca ttcttgaaga cgaaagggcc tcgtgatacg
     6481 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     6541 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     6601 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     6661 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6721 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6781 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6841 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6901 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6961 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     7021 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     7081 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     7141 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     7201 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     7261 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     7321 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     7381 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     7441 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     7501 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     7561 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     7621 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7681 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7741 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7801 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     7861 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7921 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7981 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     8041 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     8101 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     8161 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     8221 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     8281 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     8341 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     8401 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     8461 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagcaagct
     8521 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     8581 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctccccgtg
     8641 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     8701 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     8761 aattgtgagc ggataacaat ttcacacagg aaacagctat gacatgatta cgaatttcac
     8821 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     8881 ttatcatgtc tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac
     8941 cccataccct attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc
     9001 ctctgaatta ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt
     9061 agt
//
