LOCUS       sequence-432522-       11257 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Stable, inducible expression in endosome.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11257)
  AUTHORS   Zhong X, Li Q, Polacco BJ, Patil T, DiBerto JF, Vartak R, Xu J, 
            Marley A, Foussard H, Roth BL, Eckhardt M, Von Zastrow M, Krogan NJ,
            Huttenhain R
  TITLE     An automated proximity proteomics pipeline for subcellular proteome 
            and protein interaction mapping.
  JOURNAL   bioRxiv. 2023 Apr 12:2023.04.11.536358. doi: 
            10.1101/2023.04.11.536358. Preprint.
  PUBMED    37090610
REFERENCE   2  (bases 1 to 11257)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv. 
            2023 Apr 12:2023.04.11.536358. doi: 10.1101/2023.04.11.536358. 
            Preprint."
COMMENT     Sequence Label: Endo-APEX2
FEATURES             Location/Qualifiers
     source          1..11257
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(577..1368,1369..1437))
                     /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     1200..1219
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1438..1542)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    1590..1724
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             1892..2525
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    2572..2697
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3194..3427
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3612..3656
                     /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    3919..4034
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     polyA_signal    4078..4212
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4399..4425)
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             complement(4432..5178)
                     /codon_start=1
                     /product="soybean ascorbate peroxidase, engineered to serve
                     as a monomeric tag for electron microscopy and live-cell 
                     proteomics (Lam et al., 2015)"
                     /label=APEX2
                     /note="more active than the original APEX due to the A134P 
                     mutation"
                     /translation="GKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAGT
                     FDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVAV
                     EVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGHT
                     IGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVDK
                     YAADEDAFFADYAEAHQKLSELGFADA"
     CDS             complement(5179..5202)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      5202..5211
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(join(5770..6480,6481..6483,6484..6486))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(5770..6480,6481..6483,6484..6486))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5812..5833)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     6161..6180
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6420..6441
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(6496..6863)
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    6621..6639
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6657..6675
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6693..6711
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6729..6747
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6765..6783
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6801..6819
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6837..6855
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        6880..7390
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7309..7327
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7409..8155
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     promoter        8166..8495
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8346..8481
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             8504..9103
                     /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(8504..8523)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9000..9020
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9117..9705
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9170..9190)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9588..9599)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9912..10545
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10673..10689)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10673..10689)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10686..10708)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10697..10713
                     /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(10721..10727,10728..10745,10746..10751))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10766..10787
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10904..10921)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(11075..11257,1..406))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11155..11174)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
       61 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
      121 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
      181 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
      241 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
      301 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
      361 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
      421 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
      481 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
      541 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
      601 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
      661 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
      721 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
      781 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
      841 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
      901 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
      961 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     1021 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     1081 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     1141 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     1201 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     1261 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     1321 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     1381 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     1441 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     1501 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     1561 gccacctgac gtcgacggat cgggagatca acttgtttat tgcagcttat aatggttaca
     1621 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     1681 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaactgg ataactcaag
     1741 ctaaccaaaa tcatcccaaa cttcccaccc cataccctat taccactgcc aattacctgt
     1801 ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa agaaattgta
     1861 tttgttaaat atgtactaca aacttagtag ttggaagggc taattcactc ccaaagaaga
     1921 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac
     1981 tacacaccag ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta
     2041 ccagttgagc cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac
     2101 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac
     2161 agccgcctag catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc
     2221 tgatatcgag cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg
     2281 ggcgggactg gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg
     2341 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     2401 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     2461 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     2521 tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga
     2581 cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt
     2641 acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt
     2701 attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag
     2761 aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt
     2821 aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca
     2881 tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc aaccctctat
     2941 tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa
     3001 gagcaaaaca aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg
     3061 gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa
     3121 ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa
     3181 gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg
     3241 gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc
     3301 agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct
     3361 ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac
     3421 agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga
     3481 atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt
     3541 gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa
     3601 accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga
     3661 attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag
     3721 gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg
     3781 gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg
     3841 aaggaataga agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg
     3901 gatctcgacg gtatcgcctt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     3961 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     4021 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagttta tcgacttaac
     4081 ttgtttattg cagcttataa tggttacaaa taaggcaata gcatcacaaa tttcacaaat
     4141 aaggcatttt tttcactgca ttctagtttt ggtttgtcca aactcatcaa tgtatcttat
     4201 catgtctgga tctcaaatcc ctcggaagct gcgcctgtct taggttggag tgatacattt
     4261 ttatcacttt tacccgtctt tggattaggc agtagctctg acggccctcc tgtcttaggt
     4321 tagtgaaaaa tgtcactctc ttacccgtca ttggctgtcc agcttagctc gcaggggagg
     4381 tggtctggat ccttagtcca gggtcaggcg ctccagggga ggcagctgca gggcatcagc
     4441 aaacccaagc tcggaaagct tttggtgagc ctcagcgtaa tcagcaaaga aggcatcttc
     4501 gtccgctgca tatttgtcaa cgagagggcg gaatacaggg tcagacaaaa gagccttgtc
     4561 agaaggtagc tgaaggagac cttccttctc accactcaac aactccgtga agtatgagtt
     4621 gtcgaaaata agaggattag aggtccaggg accctcaaat ccagaacgct ccttgtgtgc
     4681 agctccaata gtgtgacccc cagatagagc aacgatatct tggtcagtaa gccccatagc
     4741 tttgccaaac acatctctca aatggtcaga acccttagtg ggatcgggca agcgaccctc
     4801 tggtggtggc tcaggcttgt cctctcttcc agggtggaat ggaaccttag gtccacccgt
     4861 gacctcaacg gcaacaacgc cagccaactg gtagaaatcg gcgtagctca aaatagggaa
     4921 ctccgccttg agtggctcca aaagcctaac agcgatgtca agaccgttgt tagcgctgtg
     4981 agccagttcg gcagggtgct tgatggttcc gaagggtcca ccggtcttcg tgcccttgtc
     5041 aaaggttcca gcagagtgga atgccaaacg gagcattaga ggagcgcatc tcttctcagc
     5101 gatgaagcct ctgagcttct tcttcgcctt ctcaacggcg tcctggtaat cagcactcac
     5161 agttgggtaa gactttccct tatcgtcgtc atccttgtag tccatggtgg cggccgcaga
     5221 tccggtggat cccgggcccg cggtaccgtc gacatatgcc ttcttgttca gctgctcata
     5281 gcagggctca cacacgcgca cctccttctc aatgccgaac ttggggatgg tggagtactt
     5341 ggaggagcac ttgccacaga agatctgccc acatgctcgg cagtgatgct tgcgggtcac
     5401 caccccaaac tgtactctgc accgatggca ttcctcagca tccacccagt caggggctct
     5461 ttcagcagcg aacatggcat cactttcgga tccttgtcct tgtgccttct tgttcagctg
     5521 ctcatagcag ggctcacaca cgcgcacctc cttctcaatg ccgaacttgg ggatggtgga
     5581 gtacttggag gagcacttgc cacagaagat ctgcccacat gctcggcagt gatgcttgcg
     5641 ggtcaccacc ccaaactgta ctctgcaccg atggcattcc tcagcatcca cccagtcagg
     5701 ggctctttca gcagcgaaca tggcatcact ttcgaattca attcgaagct tgagctcgag
     5761 atctgagtac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc
     5821 cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actgggtgct
     5881 caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta
     5941 gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt
     6001 gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc
     6061 cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg
     6121 gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc
     6181 cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc
     6241 gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt
     6301 cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag
     6361 cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac
     6421 gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct
     6481 caccatggtg aattctttac gagggtagga agtggtacgg aaagttggta taagacaaaa
     6541 gtgttgtgga attgaagttt actcaaaaaa tcagcactct tttataggcg ccctggttta
     6601 cataagcaaa gcttatacgt tctctatcac tgatagggag taaactggat atacgttctc
     6661 tatcactgat agggagtaaa ctgtagatac gttctctatc actgataggg agtaaactgg
     6721 tcatacgttc tctatcactg atagggagta aactccttat acgttctcta tcactgatag
     6781 ggagtaaagt ctgcatacgt tctctatcac tgatagggag taaactcttc atacgttctc
     6841 tatcactgat agggagtaaa ctcgaggtga taattccacg gggttggggt tgcgcctttt
     6901 ccaaggcagc cctgggtttg cgcagggacg cggctgctct gggcgtggtt ccgggaaacg
     6961 cagcggcgcc gaccctgggt ctcgcacatt cttcacgtcc gttcgcagcg tcacccggat
     7021 cttcgccgct acccttgtgg gccccccggc gacgcttcct gctccgcccc taagtcggga
     7081 aggttccttg cggttcgcgg cgtgccggac gtgacaaacg gaagccgcac gtctcactag
     7141 taccctcgca gacggacagc gccagggagc aatggcagcg cgccgaccgc gatgggctgt
     7201 ggccaatagc ggctgctcag cagggcgcgc cgagagcagc ggccgggaag gggcggtgcg
     7261 ggaggcgggg tgtggggcgg tagtgtgggc cctgttcctg cccgcgcggt gttccgcatt
     7321 ctgcaagcct ccggagcgca cgtcggcagt cggctccctc gttgaccgaa tcaccgacct
     7381 ctctccccag ggggatcatc gaattaccat gtctagactg gacaagagca aagtcataaa
     7441 ctctgctctg gaattactca atggagtcgg tatcgaaggc ctgacgacaa ggaaactcgc
     7501 tcaaaagctg ggagttgagc agcctaccct gtactggcac gtgaagaaca agcgggccct
     7561 gctcgatgcc ctgccaatcg agatgctgga caggcatcat acccactcct gccccctgga
     7621 aggcgagtca tggcaagact ttctgcggaa caacgccaag tcataccgct gtgctctcct
     7681 ctcacatcgc gacggggcta aagtgcatct cggcacccgc ccaacagaga aacagtacga
     7741 aaccctggaa aatcagctcg cgttcctgtg tcagcaaggc ttctccctgg agaacgcact
     7801 gtacgctctg tccgccgtgg gccactttac actgggctgc gtattggagg aacaggagca
     7861 tcaagtagca aaagaggaaa gagagacacc taccaccgat tctatgcccc cacttctgaa
     7921 acaagcaatt gagctgttcg accggcaggg agccgaacct gccttccttt tcggcctgga
     7981 actaatcata tgtggcctgg agaaacagct aaagtgcgaa agcggcgggc cgaccgacgc
     8041 ccttgacgat tttgacttag acatgctccc agccgatgcc cttgacgact ttgaccttga
     8101 tatgctgcct gctgacgctc ttgacgattt tgaccttgac atgctccccg ggtaaacgcg
     8161 cgaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     8221 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     8281 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     8341 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     8401 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     8461 gaggaggctt ttttggaggc ctaggctttt gcaaaacgcg accatgaccg agtacaagcc
     8521 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc
     8581 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt
     8641 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     8701 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     8761 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     8821 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     8881 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     8941 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     9001 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     9061 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgaacgcgtc tggaacaatc
     9121 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     9181 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     9241 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     9301 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     9361 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     9421 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     9481 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     9541 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     9601 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     9661 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg
     9721 cagtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc
     9781 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca
     9841 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     9901 aaagagggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg
     9961 gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt
    10021 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt
    10081 agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat
    10141 ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca
    10201 cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa
    10261 gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg
    10321 agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag
    10381 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
    10441 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
    10501 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
    10561 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt
    10621 gacattgcta gcgtttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
    10681 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
    10741 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
    10801 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
    10861 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
    10921 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
    10981 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
    11041 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
    11101 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
    11161 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
    11221 ccggatacct gtccgccttt ctcccttcgg gaagcgt
//
