LOCUS       Exported               10218 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses EGFP-tagged early endosome marker.
ACCESSION   .
VERSION     .
KEYWORDS    pLVX-EF1a-EGFP-RAB5A-IRES-Puromycin
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10218)
  AUTHORS   Schormann W, Hariharan S, Andrews DW
  TITLE     A reference library for assigning protein subcellular localizations 
            by image-based machine learning.
  JOURNAL   J Cell Biol. 2020 Mar 2;219(3). pii: 133635. doi: 
            10.1083/jcb.201904090.
  PUBMED    31968357
REFERENCE   2  (bases 1 to 10218)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10218
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        158..1339
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          388..1330
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1287..1307
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1394..1403
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1400..2116
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1400..2116
                     /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(1445..1466)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1706..1725)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2053..2074
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2788..3361
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2955..2972)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3182..3201
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3394..3993
                     /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(3394..3413)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3890..3910
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4007..4595
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4060..4080)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4478..4489)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4802..5435
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5563..5579)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5563..5579)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5576..5598)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5587..5603
                     /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(5611..5641)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5656..5677
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5794..5811)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5965..6553)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6045..6064)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6724..7584)
                     /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     7347..7366
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7585..7689)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    7737..7871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7774..7793)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7828..7847
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             8039..8672
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    8719..8844
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9341..9574
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9759..9803
                     /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    10066..10181
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
ORIGIN
        1 cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg ggaatcccag ggaccgtcgt
       61 taaactccca ctaacgtaga acccagagat cgctgcgttc ccgccccctc acccgcccgc
      121 tctcgtcatc actgaggtgg agaagagcat gcgtgaggct ccggtgcccg tcagtgggca
      181 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt
      241 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
      301 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
      361 cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc
      421 ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacgcccct ggctgcagta
      481 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc
      541 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cttgggcgct ggggccgccg
      601 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat
      661 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc
      721 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg
      781 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
      841 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
      901 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
      961 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg
     1021 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg
     1081 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag
     1141 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
     1201 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct
     1261 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
     1321 tccatttcag gtgtcgtgag gatctatttc cggtgaattc ctcgagacta gttctagagc
     1381 ggccgcaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1441 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1501 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     1561 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     1621 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1681 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     1741 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1801 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     1861 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     1921 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1981 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     2041 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2101 ggacgagctg tacaagtact cagatctcga catggctagt cgaggcgcaa caagacccaa
     2161 cgggccaaat actggaaata aaatatgcca gttcaaacta gtacttctgg gagagtccgc
     2221 tgttggcaaa tcaagcctag tgcttcgttt tgtgaaaggc caatttcatg aatttcaaga
     2281 gagtaccatt ggggctgctt ttctaaccca aactgtatgt cttgatgaca ctacagtaaa
     2341 gtttgaaata tgggatacag ctggtcaaga acgataccat agcctagcac caatgtacta
     2401 cagaggagca caagcagcca tagttgtata tgatatcaca aatgaggagt cctttgcaag
     2461 agcaaaaaat tgggttaaag aacttcagag gcaagcaagt cctaacattg taatagcttt
     2521 atcgggaaac aaggccgacc tagcaaataa aagagcagta gatttccagg aagcacagtc
     2581 ctatgcagat gacaatagtt tattattcat ggagacatcc gctaaaacat caatgaatgt
     2641 aaatgaaata ttcatggcaa tagctaaaaa attgccaaag aatgaaccac aaaatccagg
     2701 agcaaattct gccagaggaa gaggagtaga ccttaccgaa cccacacaac caaccaggaa
     2761 tcagtgttgt agtaactaag gatcccgccc ctctccctcc ccccccccta acgttactgg
     2821 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt
     2881 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
     2941 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
     3001 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
     3061 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
     3121 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
     3181 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
     3241 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa
     3301 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata
     3361 agcttgccac aacccacaag gagacgacct tccatgaccg agtacaagcc cacggtgcgc
     3421 ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac
     3481 taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg
     3541 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     3601 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     3661 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     3721 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     3781 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     3841 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     3901 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     3961 acctggtgca tgacccgcaa gcccggtgcc tagacgcgtc tggaacaatc aacctctgga
     4021 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     4081 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     4141 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     4201 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     4261 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     4321 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     4381 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     4441 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     4501 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     4561 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg cagtcgagac
     4621 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc
     4681 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta
     4741 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagagggga
     4801 ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac
     4861 acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca
     4921 ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc
     4981 aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg
     5041 gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga
     5101 gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc
     5161 gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga
     5221 tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg
     5281 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     5341 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     5401 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
     5461 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt gacattgcta
     5521 gcgtttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5581 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5641 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5701 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5761 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5821 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5881 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5941 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6001 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6061 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6121 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6181 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6241 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6301 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6361 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6421 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6481 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6541 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6601 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6661 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6721 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6781 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6841 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6901 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6961 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7021 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7081 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7141 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7201 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7261 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7321 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7381 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7441 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7501 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7561 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7621 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7681 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatcaact
     7741 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     7801 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     7861 atgtctggat caactggata actcaagcta accaaaatca tcccaaactt cccaccccat
     7921 accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg
     7981 aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagttg
     8041 gaagggctaa ttcactccca aagaagacaa gatatccttg atctgtggat ctaccacaca
     8101 caaggctact tccctgatta gcagaactac acaccagggc caggggtcag atatccactg
     8161 acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga agaggccaat
     8221 aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga tgacccggag
     8281 agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt ggcccgagag
     8341 ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg actttccgct
     8401 ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc cctcagatcc
     8461 tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc
     8521 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     8581 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     8641 acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg
     8701 aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca
     8761 agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag
     8821 gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa
     8881 aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca
     8941 agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt
     9001 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca
     9061 ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc
     9121 aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa
     9181 gcggccggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     9241 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     9301 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     9361 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     9421 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     9481 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     9541 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     9601 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     9661 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     9721 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     9781 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     9841 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
     9901 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
     9961 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
    10021 gagacagatc cattcgatta gtgaacggat ctcgacggta tcgcctttaa aagaaaaggg
    10081 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
    10141 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
    10201 agcagagatc cagtttat
//
