LOCUS       sequence-341832-       12047 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For lentiviral transduction of 3FLAG-EEA1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12047)
  AUTHORS   Park H, Hundley FV, Yu Q, Overmyer KA, Brademan DR, Serrano L, Paulo
            JA, Paoli JC, Swarup S, Coon JJ, Gygi SP, Wade Harper J
  TITLE     Spatial snapshots of amyloid precursor protein intramembrane 
            processing via early endosome proteomics.
  JOURNAL   Nat Commun. 2022 Oct 16;13(1):6112. doi: 10.1038/s41467-022-33881-x.
  PUBMED    36245040
REFERENCE   2  (bases 1 to 12047)
  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: "Nat 
            Commun."; date: "2022-10-16"; volume: "13(1):6112. doi"; pages: " 
            10.1038/s41467-022-33881-x"
COMMENT     Sequence Label: pHAGE-3FLAG-EEA1
FEATURES             Location/Qualifiers
     source          1..12047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2062..2179
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2231..2534
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2535..2738
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2689..2709
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2735..2759
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2835..2844
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2844..2867
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2871..2894
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2898..2921
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    2937..2961
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2967..7202
                     /codon_start=1
                     /product="human early endosome antigen 1"
                     /label=EEA1
                     /translation="MLRRILQRTPGRVGSQGSDLDSSATPINTVDVNNESSSEGFICPQ
                     CMKSLGSADELFKHYEAVHDAGNDSGHGGESNLALKRDDVTLLRQEVQDLQASLKEEKW
                     YSEELKKELEKYQGLQQQEAKPDGLVTDSSAELQSLEQQLEEAQTENFNIKQMKDLFEQ
                     KAAQLATEIADIKSKYDEERSLREAAEQKVTRLTEELNKEATVIQDLKTELLQRPGIED
                     VAVLKKELVQVQTLMDNMTLERERESEKLKDECKKLQSQYASSEATISQLRSELAKGPQ
                     EVAVYVQELQKLKSSVNELTQKNQTLTENLLKKEQDYTKLEEKHNEESVSKKNIQATLH
                     QKDLDCQQLQSRLSASETSLHRIHVELSEKGEATQKLKEELSEVETKYQHLKAEFKQLQ
                     QQREEKEQHGLQLQSEINQLHSKLLETERQLGEAHGRLKEQRQLSSEKLMDKEQQVADL
                     QLKLSRLEEQLKEKVTNSTELQHQLDKTKQQHQEQQALQQSTTAKLREAQNDLEQVLRQ
                     IGDKDQKIQNLEALLQKSKENISLLEKEREDLYAKIQAGEGETAVLNQLQEKNHTLQEQ
                     VTQLTEKLKNQSESHKQAQENLHDQVQEQKAHLRAAQDRVLSLETSVNELNSQLNESKE
                     KVSQLDIQIKAKTELLLSAEAAKTAQRADLQNHLDTAQNALQDKQQELNKITTQLDQVT
                     AKLQDKQEHCSQLESHLKEYKEKYLSLEQKTEELEGQIKKLEADSLEVKASKEQALQDL
                     QQQRQLNTDLELRATELSKQLEMEKEIVSSTRLDLQKKSEALESIKQKLTKQEEEKKIL
                     KQDFETLSQETKIQHEELNNRIQTTVTELQKVKMEKEALMTELSTVKDKLSKVSDSLKN
                     SKSEFEKENQKGKAAILDLEKTCKELKHQLQVQMENTLKEQKELKKSLEKEKEASHQLK
                     LELNSMQEQLIQAQNTLKQNEKEEQQLQGNINELKQSSEQKKKQIEALQGELKIAVLQK
                     TELENKLQQQLTQAAQELAAEKEKISVLQNNYEKSQETFKQLQSDFYGRESELLATRQD
                     LKSVEEKLSLAQEDLISNRNQIGNQNKLIQELKTAKATLEQDSAKKEQQLQERCKALQD
                     IQKEKSLKEKELVNEKSKLAEIEEIKCRQEKEITKLNEELKSHKLESIKEITNLKDAKQ
                     LLIQQKLELQGKADSLKAAVEQEKRNQQILKDQVKKEEEELKKEFIEKEAKLHSEIKEK
                     EVGMKKHEENEAKLTMQITALNENLGTVKKEWQSSQRRVSELEKQTDDLRGEIAVLEAT
                     VQNNQDERRALLERCLKGEGEIEKLQTKVLELQRKLDNTTAAVQELGRENQSLQIKHTQ
                     ALNRKWAEDNEVQNCMACGKGFSVTVRRHHCRQCGNIFCAECSAKNALTPSSKKPVRVC
                     DACFNDLQG"
     protein_bind    complement(7204..7228)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    7241..7807
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    7258..7809
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7402..7419)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7629..7648
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7821..8420
                     /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(7821..7840)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8317..8337
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8437..9025
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8490..8510)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8908..8919)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9100..9333
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9452..9470)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9538..9642
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9643..9711,9712..10503)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(9861..9880)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10674..11262
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11163..11182
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11416..11433
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11673..11694
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11709..11714,11715..11732,11733..11739)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11747..11763
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     11752..11774
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11771..11787
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11771..11787
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11837..11856
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttaattcaca
     2041 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     2101 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     2161 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggactag
     2221 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     2281 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     2341 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     2401 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     2461 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     2521 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     2581 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     2641 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     2701 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg
     2761 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccggcgg ccgcagatct
     2821 ctcgaggttg atctgccacc atggactaca aggatgacga tgacaagggc gattacaagg
     2881 atgacgacga taagggagat tacaaggatg acgacgataa ggctcgaggt tatcaaacaa
     2941 gtttgtacaa aaaagcaggc tccaccatgt taaggaggat tttacagagg actcctggga
     3001 gagttggctc tcaaggttct gatttagatt catcagcaac tcctataaac acagtggacg
     3061 tcaataatga aagctcttca gagggtttca tatgtcccca gtgtatgaaa tctcttggat
     3121 ctgctgatga acttttcaaa cattatgaag ctgttcatga tgctggtaat gactcaggtc
     3181 atggaggaga gtctaatctt gctttgaagc gagatgatgt aacactgctc agacaagagg
     3241 tccaagacct acaggcttca cttaaggaag aaaaatggta ctcggaagaa ttaaagaagg
     3301 aattagaaaa atatcaaggg ctgcagcagc aagaggccaa acctgatggg ttggtgactg
     3361 attcatcagc agaactacag tctttggaac agcaattaga agaagcccaa acagaaaatt
     3421 ttaatattaa gcaaatgaaa gacttatttg aacagaaagc agcccaactt gctactgaaa
     3481 ttgcagatat aaagtcaaag tatgatgaag aaaggagtct tcgagaagct gctgaacaaa
     3541 aagtgacacg tctgacagaa gaattaaaca aagaggcaac tgtaattcaa gatctgaaga
     3601 cggaactgct tcagagacct ggtatagaag atgttgccgt gctaaagaaa gaactggtcc
     3661 aagttcaaac actaatggat aacatgacct tggaacgtga gcgagaatct gaaaaactca
     3721 aagatgaatg caaaaaattg cagtcacaat atgctagctc agaggccaca ataagccagc
     3781 taaggagtga acttgccaaa ggcccccagg aagttgctgt atatgtacag gaactacaaa
     3841 aactgaaaag ttcagttaat gaattaacac aaaaaaatca gaccttgaca gaaaacttgc
     3901 tgaaaaaaga acaagactat actaagttag aggagaaaca taatgaagaa tctgtgagta
     3961 aaaagaatat tcaggcaacc cttcatcaaa aagacctaga ttgtcaacag cttcagtcaa
     4021 gattgtctgc atctgaaacc tcactgcata gaatacatgt agaactaagt gaaaaaggag
     4081 aagctactca aaagctcaaa gaagaattat ctgaggtaga gaccaagtac cagcatctaa
     4141 aggcggagtt taagcagcta caacaacaga gagaagaaaa ggagcagcat gggttacaac
     4201 tccaaagtga aattaatcaa ttacatagca aacttctgga gacagagcgc caactagggg
     4261 aagctcatgg taggctgaag gaacagagac agctttcaag tgaaaagttg atggataaag
     4321 aacaacaagt ggctgattta caactcaaac tttctcggtt agaagagcag ttgaaggaaa
     4381 aagttacaaa ttctacagaa ttgcagcatc aattagataa aacaaagcaa cagcatcaag
     4441 aacaacaggc tcttcagcaa agcaccacgg caaaacttcg agaagctcag aatgatttgg
     4501 aacaagttct acgtcaaatt ggcgataagg accaaaagat ccagaacctt gaagctttat
     4561 tacagaagag taaagaaaat atttcattac tagaaaaaga aagagaagat ctttatgcaa
     4621 aaattcaggc tggtgaagga gagactgctg ttcttaacca gttacaagaa aaaaaccata
     4681 cactacagga gcaagtaact caactaacag agaagctgaa gaatcagtca gaaagtcata
     4741 aacaagccca ggagaatttg catgaccagg tacaagagca gaaggcacat cttagagctg
     4801 cacaagaccg tgtcctttcc ctagaaacta gtgtcaatga attaaatagt caattaaatg
     4861 aaagcaagga gaaggtctcc cagcttgaca tacagattaa agccaaaacc gaactattac
     4921 tatcagcaga agcagcaaaa actgctcaaa gagctgatct tcagaatcat ttggacacag
     4981 ctcaaaatgc attacaagat aaacagcagg agttaaataa gattactact cagttggatc
     5041 aggtcactgc aaagttacaa gacaagcaag aacattgcag tcagctggaa agtcatctta
     5101 aagaatataa agagaaatac ctctctttag aacagaaaac cgaagagcta gaaggtcaaa
     5161 ttaagaaact agaagctgat agtcttgaag ttaaagcaag caaggagcag gctttgcaag
     5221 atctacaaca gcaaagacag ctgaacacag atttagagct cagagccaca gaattgagta
     5281 aacaacttga aatggagaag gaaatagtat ccagtacaag attggatcta cagaaaaaat
     5341 ctgaagccct tgaaagtatc aagcaaaagc ttaccaagca agaggaagaa aaaaaaatcc
     5401 tgaaacaaga ttttgaaact ttaagtcaag aaacaaagat tcagcatgag gaattgaata
     5461 acagaattca aacaacagta acagaactac aaaaagtgaa aatggagaaa gaagctttaa
     5521 tgacagagct ttctacagta aaggacaaac tatcaaaagt ttctgattct ttgaaaaact
     5581 ctaaaagtga atttgaaaag gagaatcaga aaggaaaagc cgctatatta gacttggaaa
     5641 aaacttgcaa agaattaaag catcaacttc aagtgcagat ggaaaacaca cttaaggaac
     5701 agaaggaact gaaaaagtca cttgaaaaag agaaggaggc ttctcatcag ttgaaattgg
     5761 aactcaattc aatgcaggaa caacttatac aggcccagaa tactttaaaa caaaatgaaa
     5821 aagaagagca acaacttcag gggaacataa atgagctaaa gcaatcaagt gaacagaaga
     5881 aaaaacaaat tgaagcactc caaggagagc ttaaaattgc tgttttacag aagacagagc
     5941 ttgagaataa actacagcag cagttaacac aggcagccca ggaacttgca gcagagaaag
     6001 agaaaatatc agtattacaa aacaactatg aaaaaagtca ggaaactttc aaacagcttc
     6061 aatctgattt ctatgggagg gaatctgaac ttctagccac caggcaagat cttaagtctg
     6121 tagaagagaa gctttctcta gcacaggagg acttgatttc aaacagaaat caaattggaa
     6181 atcaaaataa attgattcaa gaactgaaga ctgccaaggc tacattggag caggattcag
     6241 caaagaaaga acagcaattg caggagcgat gtaaagcact acaagacatt cagaaagaaa
     6301 agtcactgaa agaaaaagaa ctggtaaatg agaagtctaa attggcagag atagaagaaa
     6361 ttaaatgtag acaagaaaaa gaaatcacta aactaaacga agaactcaag tcccacaaac
     6421 tagaaagcat aaaggagata acaaatctta aagatgctaa acagcttcta attcagcaga
     6481 aattagaact tcaaggaaaa gcggactccc tgaaggcagc tgttgaacag gagaagagaa
     6541 atcagcagat actaaaagac caggtgaaaa aggaagaaga ggagctgaag aaagaattta
     6601 ttgagaaaga agctaagttg cattccgaaa taaaagaaaa ggaagtagga atgaagaagc
     6661 atgaagaaaa tgaggctaaa cttaccatgc agattacagc attaaatgaa aacttaggca
     6721 ctgtgaagaa ggagtggcaa tctagtcaac ggagagttag tgagcttgag aaacaaacgg
     6781 atgacttacg gggtgaaatt gcagtattag aagcaacggt tcagaataat caagatgaaa
     6841 ggagagcact actggaaaga tgtcttaaag gagaaggtga aatagaaaag cttcaaacca
     6901 aagtattaga attgcaaaga aagctggata atacaactgc agcagtgcag gagctgggca
     6961 gagaaaacca atcacttcag atcaaacata cacaagcgtt gaatagaaag tgggccgaag
     7021 acaatgaagt acaaaactgt atggcctgtg ggaaaggctt ttcagtaaca gtgagacggc
     7081 atcactgccg acagtgtgga aatatcttct gtgctgaatg ttcagccaaa aatgccttaa
     7141 ctccttcctc caagaagcct gttcgtgtct gtgatgcatg tttcaatgac ttgcaaggat
     7201 aagacccagc tttcttgtac aaagtggttc gataacgaat tccgcccccc ccccctaacg
     7261 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     7321 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     7381 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     7441 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     7501 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     7561 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     7621 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     7681 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     7741 ggttaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     7801 atgataatat ggccacaacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     7861 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     7921 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     7981 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     8041 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     8101 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     8161 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     8221 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     8281 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     8341 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     8401 cccgcaagcc cggtgcctga tgcatcgatg gatcctaatc aacctctgga ttacaaaatt
     8461 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     8521 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     8581 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     8641 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     8701 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     8761 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     8821 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     8881 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     8941 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     9001 atctcccttt gggccgcctc cccgcctgag atcctttaag accaatgact tacaaggcag
     9061 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     9121 aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     9181 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     9241 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     9301 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     9361 agtatttata acttgcaaag aaatgaatat cagagagtga gaggcccggg ttaattaagg
     9421 aaagggctag atcattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     9481 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     9541 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     9601 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     9661 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     9721 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     9781 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     9841 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     9901 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     9961 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
    10021 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
    10081 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
    10141 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
    10201 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
    10261 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
    10321 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
    10381 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
    10441 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
    10501 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
    10561 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
    10621 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
    10681 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
    10741 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
    10801 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
    10861 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
    10921 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
    10981 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
    11041 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
    11101 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
    11161 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
    11221 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
    11281 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
    11341 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
    11401 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
    11461 ccgcctctcc ccgcgcgttg gccgattcat taatgcagca agctcatggc tgactaattt
    11521 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
    11581 gaggcttttt tggaggccta ggcttttgca aaaagctccc cgtggcacga caggtttccc
    11641 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
    11701 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
    11761 caatttcaca caggaaacag ctatgacatg attacgaatt tcacaaataa agcatttttt
    11821 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
    11881 aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc
    11941 actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga attattttca
    12001 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagt
//
