LOCUS       sequence-386451-        9404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  As mBFP-APP-mGFP, but with substitution at position 612 that 
            prevents alpha-secretase activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9404)
  AUTHORS   Capitini C, Bigi A, Parenti N, Emanuele M, Bianchi N, Cascella R, 
            Cecchi C, Maggi L, Annunziato F, Pavone FS, Calamai M
  TITLE     APP and Bace1: Differential effect of cholesterol enrichment on 
            processing and plasma membrane mobility.
  JOURNAL   iScience. 2023 Apr 11;26(5):106611. doi: 10.1016/j.isci.2023.106611.
            eCollection 2023 May 19.
  PUBMED    37128606
REFERENCE   2  (bases 1 to 9404)
  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"; journalName: "iScience.";
            date: "2023-04-11"; volume: "26(5):106611. doi"; pages: " 
            10.1016/j.isci.2023.106611. eCollection 2023 May 19"
COMMENT     Sequence Label: mBFP-APPP1-mGFP
FEATURES             Location/Qualifiers
     source          1..9404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     165..182
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     877..897
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     952..971
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1086..1784
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             complement(2082..2093)
                     /codon_start=1
                     /gene="cspA"
                     /product="translation enhancing element for E. coli (Qing 
                     et al., 2004)"
                     /label=TEE
                     /translation="NHKV"
     CDS             2573..2593
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             3861..4574
                     /codon_start=1
                     /product="monomeric green fluorescent protein, also known 
                     as mTagGFP"
                     /label=TagGFP2
                     /note="mammalian codon-optimized"
                     /translation="MSGGEELFAGIVPVLIELDGDVHGHKFSVRGEGEGDADYGKLEIK
                     FICTTGKLPVPWPTLVTTLCYGIQCFARYPEHMKMNDFFKSAMPEGYIQERTIQFQDDG
                     KYKTRGEVKFEGDTLVNRIELKGKDFKEDGNILGHKLEYSFNSHNVYIRPDKANNGLEA
                     NFKTRHNIEGGGVQLADHYQTNVPLGDGPVLIPINHYLSTQTKISKDRNEARDHMVLLE
                     SFSACCHTHGMDELYR"
     primer_bind     complement(4609..4625)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    4636..5258
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4835..4854)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(5282..5302)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5287..5616
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5467..5602
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5683..6477
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5737..5756)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6347..6366
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6651..6772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6688..6707)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6742..6761
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6821..6837)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6834..6856)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6845..6861
                     /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(6869..6875,6876..6893,6894..6899))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6914..6935
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7052..7069)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7223..7811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7303..7322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7982..8773,8774..8842))
                     /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     8605..8624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8843..8947)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8974..9402
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9061..9080)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9271..9292
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg
     1021 cgatcgccat gctgcccggt ttggcactgc tcctgctggc cgcctggacg gctcgggcgg
     1081 ctagcatgag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg
     1141 tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc
     1201 agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
     1261 ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc atccccgact
     1321 tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg
     1381 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg
     1441 tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag aaaacactcg
     1501 gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa ggcagaaacg
     1561 acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacgccaag accacatata
     1621 gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg gactacagac
     1681 tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag gtggcagtgg
     1741 ccagatactg cgacctccct agcaaactgg ggcacaagct taatagtcgt ggaacaggcg
     1801 cgcctctgga ggtacccact gatggtaatg ctggcctgct ggctgaaccc cagattgcca
     1861 tgttctgtgg cagactgaac atgcacatga atgtccagaa tgggaagtgg gattcagatc
     1921 catcagggac caaaacctgc attgatacca aggaaggcat cctgcagtat tgccaagaag
     1981 tctaccctga actgcagatc accaatgtgg tagaagccaa ccaaccagtg accatccaga
     2041 actggtgcaa gcggggccgc aagcagtgca agacccatcc ccactttgtg attccctacc
     2101 gctgcttagt tggtgagttt gtaagtgatg cccttctcgt tcctgacaag tgcaaattct
     2161 tacaccagga gaggatggat gtttgcgaaa ctcatcttca ctggcacacc gtcgccaaag
     2221 agacatgcag tgagaagagt accaacttgc atgactacgg catgttgctg ccctgcggaa
     2281 ttgacaagtt ccgaggggta gagtttgtgt gttgcccact ggctgaagaa agtgacaatg
     2341 tggattctgc tgatgcggag gaggatgact cggatgtctg gtggggcgga gcagacacag
     2401 actatgcaga tgggagtgaa gacaaagtag tagaagtagc agaggaggaa gaagtggctg
     2461 aggtggaaga agaagaagcc gatgatgacg aggacgatga ggatggtgat gaggtagagg
     2521 aagaggctga ggaaccctac gaagaagcca cagagagaac caccagcatt gccaccacca
     2581 ccaccaccac cacagagtct gtggaagagg tggttcgagt tcctacaaca gcagccagta
     2641 cccctgatgc cgttgacaag tatctcgaga cacctgggga tgagaatgaa catgcccatt
     2701 tccagaaagc caaagagagg cttgaggcca agcaccgaga gagaatgtcc caggtcatga
     2761 gagaatggga agaggcagaa cgtcaagcaa agaacttgcc taaagctgat aagaaggcag
     2821 ttatccagca tttccaggag aaagtggaat ctttggaaca ggaagcagcc aacgagagac
     2881 agcagctggt ggagacacac atggccagag tggaagccat gctcaatgac cgccgccgcc
     2941 tggccctgga gaactacatc accgctctgc aggctgttcc tcctcggcct cgtcacgtgt
     3001 tcaatatgct aaagaagtat gtccgcgcag aacagaagga cagacagcac accctaaagc
     3061 atttcgagca tgtgcgcatg gtggatccca agaaagccgc tcagatccgg tcccaggtta
     3121 tgacacacct ccgtgtgatt tatgagcgca tgaatcagtc tctctccctg ctctacaacg
     3181 tgcctgcagt ggccgaggag attcaggatg aagttgatga gctgcttcag aaagagcaaa
     3241 actattcaga tgacgtcttg gccaacatga ttagtgaacc aaggatcagt tacggaaacg
     3301 atgctctcat gccatctttg accgaaacga aaaccaccgt ggagctcctt cccgtgaatg
     3361 gagagttcag cctggacgat ctccagccgt ggcattcttt tggggctgac tctgtgccag
     3421 ccaacacaga aaacgaagtt gagcctgttg atgcccgccc tgctgccgac cgaggactga
     3481 ccactcgacc aggttctggg ttgacaaata tcaagacgga ggagatctct gaagtgaaga
     3541 tggatgcaga attccgacat gactcaggat atgaagttca tcatcaagta ttggtgttct
     3601 ttgcagaaga tgtgggttca aacaaaggtg caatcattgg actcatggtg ggcggtgttg
     3661 tcatagcgac agtgatcgtc atcaccttgg tgatgctgaa gaagaaacag tacacatcca
     3721 ttcatcatgg tgtggtggag gttgacgccg ctgtcacccc agaggagcgc cacctgtcca
     3781 agatgcagca gaacggctac gaaaatccaa cctacaagtt ctttgagcag atgcagaaca
     3841 cgcgtacgcg gccgctcgag atgagcgggg gcgaggagct gttcgccggc atcgtgcccg
     3901 tgctgatcga gctggacggc gacgtgcacg gccacaagtt cagcgtgcgc ggcgagggcg
     3961 agggcgacgc cgactacggc aagctggaga tcaagttcat ctgcaccacc ggcaagctgc
     4021 ccgtgccctg gcccaccctg gtgaccaccc tctgctacgg catccagtgc ttcgcccgct
     4081 accccgagca catgaagatg aacgacttct tcaagagcgc catgcccgag ggctacatcc
     4141 aggagcgcac catccagttc caggacgacg gcaagtacaa gacccgcggc gaggtgaagt
     4201 tcgagggcga caccctggtg aaccgcatcg agctgaaggg caaggacttc aaggaggacg
     4261 gcaacatcct gggccacaag ctggagtaca gcttcaacag ccacaacgtg tacatccgcc
     4321 ccgacaaggc caacaacggc ctggaggcta acttcaagac ccgccacaac atcgagggcg
     4381 gcggcgtgca gctggccgac cactaccaga ccaacgtgcc cctgggcgac ggccccgtgc
     4441 tgatccccat caaccactac ctgagcactc agaccaagat cagcaaggac cgcaacgagg
     4501 cccgcgacca catggtgctc ctggagtcct tcagcgcctg ctgccacacc cacggcatgg
     4561 acgagctgta caggtccgga ctcagataag tttaaacggc cggccgcggt catagctgtt
     4621 tcctgaacag atcccgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     4681 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     4741 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     4801 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     4861 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     4921 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     4981 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     5041 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     5101 ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca gacacagcat
     5161 aggctacctg gccatgccca accggtggga catttgagtt gcttgcttgg cactgtcctc
     5221 tcatgcgttg ggtccactca gtagatgcct gttgaattgg gtacgcggcc agcttggctg
     5281 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5341 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5401 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5461 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5521 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     5581 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     5641 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     5701 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5761 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5821 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5881 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5941 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     6001 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     6061 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     6121 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     6181 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     6241 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     6301 atcgactgtg gccggctggg tgtggccgac cgctatcagg acatagcgtt ggctacccgt
     6361 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     6421 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     6481 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     6541 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     6601 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     6661 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     6721 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     6781 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     6841 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6901 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6961 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     7021 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     7081 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     7141 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     7201 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     7261 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     7321 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     7381 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     7441 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7501 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7561 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7621 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     7681 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7741 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7801 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7861 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7921 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7981 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     8041 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     8101 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     8161 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     8221 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     8281 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     8341 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     8401 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     8461 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     8521 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8581 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8641 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8701 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8761 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8821 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8881 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8941 ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg
     9001 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     9061 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     9121 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     9181 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     9241 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     9301 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     9361 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa tttt
//
