PCDH-TetO-mEGFP-TRIM6-TurboID

LOCUS       PCDH-TetO-mEGFP-TRIM6-TurboID              9641 bp DNA     circular SYN 04-JUL-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9641)
  AUTHORS   15129015045
  TITLE     Direct Submission
  JOURNAL   Exported Aug 12, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..9641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             234..414
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    458..583
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1076..1309
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1798..1915
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    1980..2250
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2283..2320
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             2444..3160
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      2444..2447
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3161..3190
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    3197..4660
                     /label=TRIM6
     CDS             4667..4693
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4697..4723
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4727..4753
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4754..5734
                     /codon_start=1
                     /label=TurboID
                     /translation="ASKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLR
                     DWGVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKS
                     GDACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEKEFSRAD"
     misc_feature    5738..6325
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6209..6220)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6398..6631
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6703..6824
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6843..6978
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7016..7032)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7040..7056
                     /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)."
     promoter        complement(7064..7094)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7109..7130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7418..8006)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8177..9037)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9038..9142)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9616..9632
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtcgg gtttattaca gggacagcag agatccagtt tggttatcga gtttaccact
     1981 ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga
     2041 aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagttt
     2101 accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga
     2161 tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc
     2221 gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagctcgg tacccgggtc
     2281 gagtaggcgt gtacggtggg aggcctatat aagcagagct cgtttagtga accgtcagat
     2341 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
     2401 cctccgcggc ccctctagac cgctagcgct accggtcgcc accatggtga gcaagggcga
     2461 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2521 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2581 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2641 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2701 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     2761 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2821 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     2881 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     2941 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     3001 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     3061 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     3121 cgccgccggg atcactctcg gcatggacga gctgtacaag tccggactca gatctcgagc
     3181 tcaagcttcg gaattcatga cttcaccagt actggtggac atacgagaag aggtgacctg
     3241 ccctatctgc ctggagctcc taacagaacc cctgagcata gactgtggcc acagcttctg
     3301 ccaagcctgc atcacaccaa atggcaggga atcagtgatt ggtcaagaag gggaaagaag
     3361 ctgccctgtg tgccagacca gctaccagcc agggaacctg cggcctaatc ggcatctggc
     3421 caacatagtg aggcggctca gagaggtagt gttgggccct gggaagcagc tgaaagcagt
     3481 tctttgtgca gaccatggag aaaaactgca gctcttctgt caggaggatg ggaaggtcat
     3541 ttgctggctt tgtgagcggt ctcaggagca ccgtggtcac cacacgttcc tcgtggagga
     3601 ggttgcccag gagtaccagg agaagtttca ggagtctcta aagaagctga agaacgagga
     3661 gcaggaagct gagaagctaa cagcttttat cagagagaag aaaacatcct ggaagaatca
     3721 gatggagcct gagagatgca ggatccagac agagtttaat cagctgcgaa atatcctaga
     3781 cagagtggag caacgggagc tgaaaaagct ggaacaggaa gagaagaagg ggctacgaat
     3841 tatagaagag gctgagaatg atctggtcca ccagacccag tcgctgcgag agctcatctc
     3901 ggatctggag cgtcgatgtc aggggtcaac aatggagctg ctgcaggatg tgagtgatgt
     3961 cacagaaagg agtgagttct ggaccctgag gaagccagaa gctctcccta caaagctgag
     4021 aagtatgttc cgagccccag atctgaaaag gatgctgcga gtgtgtagag agctgacaga
     4081 tgtccaaagc tactgggttg acgtgaccct gaatccacac acagctaatt taaatcttgt
     4141 cctggctaaa aaccggagac aagtgaggtt tgtgggagct aaagtatctg gaccttcctg
     4201 tctggaaaag cattatgact gtagtgtcct gggctcccag cacttctcct ctggtaagca
     4261 ttactgggag gtagatgtgg ccaagaagac tgcctggatc ctgggggtat gcagcaattc
     4321 actgggacct acattctctt tcaaccattt tgctcaaaat cacagtgctt actccaggta
     4381 tcagcctcag agtggatact gggtgattgg gttacagcat aaccatgaat atagggccta
     4441 tgaggattct tccccttccc tgcttctctc catgacagtg ccccctcgcc gtgttggggt
     4501 tttcttagat tatgaggctg gtactgtctc cttttataat gtcacaaacc atggcttccc
     4561 catctacact ttctctaaat attactttcc cactactctt tgtccatatt ttaatccttg
     4621 caactgtgta attcctatga ccctgcgtcg tccaagctct ggatcctacc cgtatgatgt
     4681 tccggattac gctggctatc cctacgacgt gcccgactat gccgggtacc cctatgacgt
     4741 cccagactac gcagctagca aagacaatac tgtgcctctg aagctgatcg ctctcctggc
     4801 taatggcgag ttccatagtg gcgaacagct gggagaaacc ctgggcatgt ccagggccgc
     4861 tatcaacaag cacattcaga ctctgcgcga ctggggcgtg gacgtgttca ccgtgcccgg
     4921 aaagggctac tctctgcccg agcctatccc gctgctgaac gctaaacaga ttctgggaca
     4981 gctggacggc gggagcgtgg cagtcctgcc tgtggtcgac tccaccaatc agtacctgct
     5041 ggatcgaatc ggcgagctga agagtgggga tgcttgcatt gcagaatatc agcaggcagg
     5101 gagaggaagc agagggagga aatggttctc tccttttgga gctaacctgt acctgagtat
     5161 gttttggcgc ctgaagcggg gaccagcagc aatcggcctg ggcccggtca tcggaattgt
     5221 catggcagaa gcgctgcgaa agctgggagc agacaaggtg cgagtcaaat ggcccaatga
     5281 cctgtatctg caggatagaa agctggcagg catcctggtg gagctggccg gaataacagg
     5341 cgatgctgca cagatcgtca ttggcgccgg gattaacgtg gctatgaggc gcgtggagga
     5401 aagcgtggtc aatcagggct ggatcacact gcaggaagca gggattaacc tggacaggaa
     5461 tactctggcc gctacgctga tccgagagct gcgggcagcc ctggaactgt tcgagcagga
     5521 aggcctggct ccatatctgc cacggtggga gaagctggat aacttcatca atagacccgt
     5581 gaagctgatc attggggaca aagagatttt cgggattagc cgggggattg ataaacaggg
     5641 agccctgctg ctggaacagg acggagttat caaaccctgg atgggcggag aaatcagtct
     5701 gcggtctgcc gaaaaggagt tcagcagggc cgactgaaat caacctctgg attacaaaat
     5761 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5821 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5881 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5941 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     6001 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     6061 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     6121 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     6181 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     6241 cggcctgctg ccggctctgc ggcctcttcc gcgtctccgc cttcgccctc agacgagtcg
     6301 gatctccctt tggccgcctc cccgccggta cctttaagac caatgactta caaggcagct
     6361 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     6421 cgaaaataag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     6481 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     6541 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     6601 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     6661 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     6721 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6781 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     6841 ctatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6901 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     6961 gaggcttttt tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat
     7021 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     7081 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     7141 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     7201 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     7261 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     7321 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     7381 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     7441 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7501 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7561 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     7621 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7681 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7741 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7801 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     7861 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7921 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7981 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     8041 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     8101 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     8161 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     8221 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     8281 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     8341 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     8401 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     8461 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     8521 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     8581 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     8641 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     8701 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8761 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8821 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8881 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8941 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     9001 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     9061 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     9121 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     9181 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     9241 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     9301 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     9361 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     9421 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     9481 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     9541 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     9601 cccagtcacg acgttgtaaa acgacggcca gtgccaagct g
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