LOCUS       sequence-489517-        9951 bp    DNA     circular SYN 16-OCT-2025
DEFINITION  EF-1alpha driven EV reporter E-'No'Mi-R, a re-engineered tetraspanin
            scaffold tagged with bioluminescent and fluorescent reporter 
            proteins. Nanoluc and Flag tag are removable with TEVp..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9951)
  AUTHORS   Obuchi W, Zargani-Piccardi A, Leandro K, Rufino-Ramos D, Di Lanni E,
            Frederick DM, Maalouf K, Nieland L, Xiao T, Repiton P, Vaine CA, 
            Kleinstiver BP, Bragg DC, Lee H, Miller MA, Breakefield XO, Breyne K
  TITLE     Engineering of CD63 Enables Selective Extracellular Vesicle Cargo 
            Loading and Enhanced Payload Delivery.
  JOURNAL   J Extracell Vesicles. 2025 Jun;14(6):e70094. doi: 
            10.1002/jev2.70094.
  PUBMED    40527733
REFERENCE   2  (bases 1 to 9951)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Extracell
            Vesicles."; date: "2025-06"; volume: "14(6):e70094. doi"; pages: " 
            10.1002/jev2.70094"
COMMENT     Sequence Label: pCDH-EF1-E-'No'Mi-Red-T2A-Puro
FEATURES             Location/Qualifiers
     source          1..9951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(493..1284,1285..1353))
                     /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"
     primer_bind     1116..1135
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1354..1458)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1526..1544
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1582..1604)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1704..1723
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1917..1939
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1931..1948
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1932..1948
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1963..2189
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2190..2370
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2417..2542
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3035..3268
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3453..3497
                     /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    3764..3880
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4000..5178
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4231..5169
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5126..5146
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             5338..5358
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5359..5424
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5458..5970
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             5971..5991
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             6601..7308
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(6746..6764)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7014..7033
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             7321..7374
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             7375..7974
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7375..7394)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7871..7891
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7987..8575
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8040..8060)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8458..8469)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8649..8882
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8954..9075
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8991..9010)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9045..9064
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9115..9250
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9283..9299)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9283..9299)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9296..9318)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9307..9323
                     /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(join(9331..9337,9338..9355,9356..9361))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9376..9397
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9514..9531)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(9685..9951,1..322))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
       61 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
      121 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
      181 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
      241 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
      301 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
      361 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
      421 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
      481 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
      541 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
      601 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
      661 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
      721 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
      781 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
      841 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
      901 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
      961 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     1021 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     1081 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     1141 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     1201 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     1261 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     1321 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     1381 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     1441 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     1501 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     1561 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     1621 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     1681 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     1741 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     1801 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     1861 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     1921 gtcacgacgt tgtaaaacga cggccagtgc caagctgacg cgtgtagtct tatgcaatac
     1981 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
     2041 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
     2101 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
     2161 tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct
     2221 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     2281 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     2341 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
     2401 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     2461 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     2521 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     2581 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     2641 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     2701 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     2761 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     2821 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     2881 ggccactgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     2941 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     3001 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     3061 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     3121 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     3181 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     3241 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     3301 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     3361 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     3421 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     3481 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     3541 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     3601 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     3661 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     3721 gatccattcg attagtgaac ggatctcgac ggtatcggtt aacttttaaa agaaaagggg
     3781 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     3841 ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt atcgatatcg atagctatcg
     3901 atatcgatat cgatagcttt gcaaagatgg ataaagtttt aaacagagag gaatctttgc
     3961 agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg
     4021 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc
     4081 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     4141 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     4201 tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct
     4261 ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac
     4321 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct
     4381 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc
     4441 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt
     4501 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg
     4561 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt
     4621 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
     4681 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
     4741 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
     4801 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg
     4861 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
     4921 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt
     4981 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
     5041 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt
     5101 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
     5161 ccatttcagg tgtcgtgagg aattggatcc accatggcgg tggaaggagg aatgaaatgt
     5221 gtgaagttct tgctctacgt cctcctgctg gccttttgcg cctgtgcagt gggactgatt
     5281 gccgtgggtg tcggggcaca gcttgtcctg agtcagacca tatctagaat tcctagggag
     5341 aacctctact ttcagtccga ctacaaagac catgacggtg attataaaga tcatgacatc
     5401 gattacaagg atgacgatga caagggtggt tctggtggtg gttctggtcg atccaccatg
     5461 gtcttcacac tcgaagattt cgttggggac tggcgacaga cagccggcta caacctggac
     5521 caagtccttg aacagggagg tgtgtccagt ttgtttcaga atctcggggt gtccgtaact
     5581 ccgatccaaa ggattgtcct gagcggtgaa aatgggctga agatcgacat ccatgtcatc
     5641 atcccgtatg aaggtctgag cggcgaccaa atgggccaga tcgaaaaaat ttttaaggtg
     5701 gtgtaccctg tggatgatca tcactttaag gtgatcctgc actatggcac actggtaatc
     5761 gacggggtta cgccgaacat gatcgactat ttcggacggc cgtatgaagg catcgccgtg
     5821 ttcgacggca aaaagatcac tgtaacaggg accctgtgga acggcaacaa aattatcgac
     5881 gagcgcctga tcaaccccga cggctccctg ctgttccgag taaccatcaa cggagtgacc
     5941 ggctggcggc tgtgcgaacg cattctggcg gagaacctct actttcagtc cggcgcgcct
     6001 tctagaatcc agggggctac ccctggctct ctgttgccag tggtcatcat cgcagtgggt
     6061 gtcttcctct tcctggtggc ttttgtgggc tgctgcgggg cctgcaagga gaactattgt
     6121 cttatgatca cgtttgccat ctttctgtct cttatcatgt tggtggaggt ggccgcagcc
     6181 attgctggct atgtgtttag agataaggtg atgtcagagt ttaataacaa cttccggcag
     6241 cagatggaga attacccgaa aaataaccac actgcttcga tcctggacag gatgcaggca
     6301 gattttaagt gctgtggggc tgctaactac acagattggg agaaaatccc ttccatgtcg
     6361 aagaaccgag tccccgactc ctgctgcatt aatgttactg tgggctgtgg gattaatttc
     6421 aacgagaagg cgatccataa ggagggctgt gtggagaaga ttgggggctg gctgaggaaa
     6481 aatgtgctgg tggtagctgc agcagccctt ggaattgctt ttgtcgaggt tttgggaatt
     6541 gtctttgcct gctgccttgt gaagagtatc agaagtggct acgaggtgat ggaattctgt
     6601 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
     6661 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
     6721 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
     6781 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
     6841 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
     6901 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
     6961 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
     7021 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
     7081 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
     7141 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
     7201 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
     7261 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta cggcagtgga
     7321 gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg cccaatgacc
     7381 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     7441 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     7501 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     7561 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     7621 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     7681 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     7741 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     7801 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     7861 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     7921 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgagaattc
     7981 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     8041 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     8101 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     8161 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     8221 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     8281 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     8341 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     8401 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     8461 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     8521 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctggt
     8581 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     8641 ggggggactg gaagggctaa ttcactccca acgaagataa gatctgcttt ttgcttgtac
     8701 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     8761 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     8821 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     8881 cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc
     8941 agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     9001 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     9061 catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg
     9121 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     9181 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     9241 tttggaggcc tagacttttg cagagaccaa attcgtaatc atgtcatagc tgtttcctgt
     9301 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     9361 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     9421 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     9481 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     9541 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     9601 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     9661 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     9721 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     9781 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     9841 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     9901 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc c
//
