LOCUS       sequence-419158-       10881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E-NoMi is a re-engineered tetraspanin scaffold tagged with 
            bioluminescent and fluorescent reporter proteins under an EF-1-alpha
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10881)
  AUTHORS   Maalouf KE, Vaine CA, Frederick DM, Yoshinaga A, Obuchi W, Mahjoum 
            S, Nieland L, Al Ali J, Bragg DC, Breakefield XO, Breyne K
  TITLE     Tracking human neurologic disease status in mouse brain/plasma using
            reporter-tagged, EV-associated biomarkers.
  JOURNAL   Mol Ther. 2023 Jul 5;31(7):2206-2219. doi: 
            10.1016/j.ymthe.2023.05.011. Epub 2023 May 16.
  PUBMED    37198883
REFERENCE   2  (bases 1 to 10881)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.ymthe.2023.05.011"; journalName: "Mol Ther"; date: 
            "2023-07-5- 5"; volume: "31"; issue: "7"; pages: "2206-2219"
COMMENT     Sequence Label: pCDH-EF1-E-NoMi-P2A-CopGFP-T2A-PuroR
FEATURES             Location/Qualifiers
     source          1..10881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..232
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             233..413
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    460..585
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1078..1311
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1496..1540
                     /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    1807..1923
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2043..3221
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2274..3212
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3169..3189
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3372..3437
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3471..3983
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     misc_feature    4581..4661
                     /label=MCS
                     /note="multiple cloning site"
     CDS             4662..5369
                     /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(4807..4825)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5075..5094
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    5370..5450
                     /label=MCS
                     /note="multiple cloning site"
     CDS             5466..5522
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5547..6209
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
     CDS             6294..6347
                     /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             6348..6947
                     /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(6348..6367)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6844..6864
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6960..7548
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7013..7033)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7431..7442)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7622..7855
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7927..8048
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7964..7983)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8018..8037
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8088..8223
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8256..8272)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8256..8272)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8269..8291)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8280..8296
                     /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(8304..8310,8311..8328,8329..8334))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8349..8370
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8487..8504)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8658..9246)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(9417..10208,10209..10277))
                     /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     10040..10059
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10278..10382)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     10450..10468
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(10506..10528)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     10628..10647
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10841..10863
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10855..10872
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     10856..10872
                     /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 cgatacataa acgggtctct
      241 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
      301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
      361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
      421 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
      481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
      541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
      601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
      661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
      721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
      781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
      841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
      901 agtaagacca ccgcacagca agcggccact gatcttcaga cctggaggag gagatatgag
      961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
     1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
     1801 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     1861 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aattcaaaat
     1921 tttatcgata tcgatagcta tcgatatcga tatcgatagc tttgcaaaga tggataaagt
     1981 tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa
     2041 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     2101 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     2161 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     2221 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
     2281 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     2341 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
     2401 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
     2461 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
     2521 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
     2581 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
     2641 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
     2701 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
     2761 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     2821 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     2881 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     2941 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     3001 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     3061 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     3121 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     3181 cagtggttca aagttttttt cttccatttc aggtgtcgtg aggaattgga tccaccatgg
     3241 cggtggaagg aggaatgaaa tgtgtgaagt tcttgctcta cgtcctcctg ctggcctttt
     3301 gcgcctgtgc agtgggactg attgccgtgg gtgtcggggc acagcttgtc ctgagtcaga
     3361 ccatatctag agactacaaa gaccatgacg gtgattataa agatcatgac atcgattaca
     3421 aggatgacga tgacaagggt ggttctggtg gtggttctgg tcgatccacc atggtcttca
     3481 cactcgaaga tttcgttggg gactggcgac agacagccgg ctacaacctg gaccaagtcc
     3541 ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg ggtgtccgta actccgatcc
     3601 aaaggattgt cctgagcggt gaaaatgggc tgaagatcga catccatgtc atcatcccgt
     3661 atgaaggtct gagcggcgac caaatgggcc agatcgaaaa aatttttaag gtggtgtacc
     3721 ctgtggatga tcatcacttt aaggtgatcc tgcactatgg cacactggta atcgacgggg
     3781 ttacgccgaa catgatcgac tatttcggac ggccgtatga aggcatcgcc gtgttcgacg
     3841 gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa caaaattatc gacgagcgcc
     3901 tgatcaaccc cgacggctcc ctgctgttcc gagtaaccat caacggagtg accggctggc
     3961 ggctgtgcga acgcattctg gcgtctagaa tccagggggc tacccctggc tctctgttgc
     4021 cagtggtcat catcgcagtg ggtgtcttcc tcttcctggt ggcttttgtg ggctgctgcg
     4081 gggcctgcaa ggagaactat tgtcttatga tcacgtttgc catctttctg tctcttatca
     4141 tgttggtgga ggtggccgca gccattgctg gctatgtgtt tagagataag gtgatgtcag
     4201 agtttaataa caacttccgg cagcagatgg agaattaccc gaaaaataac cacactgctt
     4261 cgatcctgga caggatgcag gcagatttta agtgctgtgg ggctgctaac tacacagatt
     4321 gggagaaaat cccttccatg tcgaagaacc gagtccccga ctcctgctgc attaatgtta
     4381 ctgtgggctg tgggattaat ttcaacgaga aggcgatcca taaggagggc tgtgtggaga
     4441 agattggggg ctggctgagg aaaaatgtgc tggtggtagc tgcagcagcc cttggaattg
     4501 cttttgtcga ggttttggga attgtctttg cctgctgcct tgtgaagagt atcagaagtg
     4561 gctacgaggt gatggaattc gctagcgcta ccggactcag atctcgagct caagcttcga
     4621 attctgcagt cgacggtacc gcgggcccgg gatccaccgg tatggtgagc aagggcgagg
     4681 aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg
     4741 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     4801 agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt
     4861 cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact
     4921 acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     4981 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg
     5041 tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg
     5101 gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca
     5161 agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca
     5221 aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca
     5281 cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact
     5341 ccaccggcgg catggacgag ctgtacaagg ctagcgctac cggactcaga tctcgagctc
     5401 aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccggt aagcttggaa
     5461 gcggagctac taacttcagc ctgctgaagc aggctggaga cgtggaggag aaccctggac
     5521 ctatggagag cgacgagagc ggcctgcccg ccatggagat cgagtgccgc atcaccggca
     5581 ccctgaacgg cgtggagttc gagctggtgg gcggcggaga gggcaccccc aagcagggcc
     5641 gcatgaccaa caagatgaag agcaccaaag gcgccctgac cttcagcccc tacctgctga
     5701 gccacgtgat gggctacggc ttctaccact tcggcaccta ccccagcggc tacgagaacc
     5761 ccttcctgca cgccatcaac aacggcggct acaccaacac ccgcatcgag aagtacgagg
     5821 acggcggcgt gctgcacgtg agcttcagct accgctacga ggccggccgc gtgatcggcg
     5881 acttcaaggt ggtgggcacc ggcttccccg aggacagcgt gatcttcacc gacaagatca
     5941 tccgcagcaa cgccaccgtg gagcacctgc accccatggg cgataacgtg ctggtgggca
     6001 gcttcgcccg caccttcagc ctgcgcgacg gcggctacta cagcttcgtg gtggacagcc
     6061 acatgcactt caagagcgcc atccacccca gcatcctgca gaacgggggc cccatgttcg
     6121 ccttccgccg cgtggaggag ctgcacagca acaccgagct gggcatcgtg gagtaccagc
     6181 acgccttcaa gacccccatt gccttcgcca gatcccgcgc tcagtcgtcc aattctgccg
     6241 tggacggcac cgccggaccc ggctccaccg gatctcgcgt cgacggcagt ggagagggca
     6301 gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc tggcccaatg accgagtaca
     6361 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     6421 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     6481 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     6541 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     6601 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     6661 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     6721 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     6781 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     6841 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     6901 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgagaa ttcgtcgaca
     6961 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     7021 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     7081 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     7141 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     7201 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     7261 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     7321 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     7381 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     7441 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     7501 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta
     7561 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     7621 ctggaagggc taattcactc ccaacgaaga taagatctgc tttttgcttg tactgggtct
     7681 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     7741 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     7801 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     7861 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     7921 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     7981 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     8041 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa
     8101 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     8161 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     8221 gcctagactt ttgcagagac caaattcgta atcatgtcat agctgtttcc tgtgtgaaat
     8281 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     8341 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     8401 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     8461 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     8521 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     8581 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     8641 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     8701 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     8761 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     8821 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     8881 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     8941 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     9001 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     9061 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     9121 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     9181 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     9241 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     9301 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     9361 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     9421 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     9481 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     9541 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     9601 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     9661 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     9721 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     9781 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     9841 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     9901 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     9961 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
    10021 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
    10081 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
    10141 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
    10201 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
    10261 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
    10321 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
    10381 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
    10441 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
    10501 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
    10561 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
    10621 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
    10681 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
    10741 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
    10801 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
    10861 acgacggcca gtgccaagct g
//
