LOCUS       sequence-490945-        9620 bp    DNA     circular SYN 28-OCT-2025
DEFINITION  HA-tagged human RhoA(G14V) cloned into the pCW backbone (Addgene 
            #41393, with puromycin resistance) together with an IRES GFP to 
            indicate doxycycline-inducible expression by fluorescence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9620)
  TITLE     Doxycycline-inducible pCW vectors with HA-tagged genes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9620)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 28, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pCW-Rho(G14V)-HA
FEATURES             Location/Qualifiers
     source          1..9620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    41..158
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        212..526
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    220..238
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    256..274
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    291..309
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    327..345
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    363..381
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    398..416
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    434..452
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     499..523
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      543..552
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             549..1127
                     /codon_start=1
                     /product="human Rho family small GTPase RhoA"
                     /label=RhoA
                     /translation="MAAIRKKLVIVGDVACGKTCLLIVFSKDQFPEVYVPTVFENYVAD
                     IEVDGKQVELALWDTAGQEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVK
                     HFCPNVPIILVGNKKDLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKT
                     KDGVREVFEMATRAALQARRGKKKSGCLVL"
     CDS             1131..1157
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1161..1187
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1256..1822
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1272..1824
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1416..1433)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1643..1662
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1836..1838,1839..1841,1842..2555)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1836..1838,1839..1841,1842..2552)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1881..1902)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2142..2161)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2489..2510
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        2562..3072
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2991..3009
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3082..3678
                     /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(3082..3101)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3578..3598
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             3679..3732
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(3733..4350,4351..4356,4357..4392,4393..4395,4396..4431,
                     4432..4434,4435..4470,4471..4479)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(4519..4539)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4937..4948)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5153..5386
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5458..5579
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5495..5514)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5549..5568
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5619..5754
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5754..5825
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5826..5894,5895..6686)
                     /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     complement(6044..6063)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6857..7445
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7346..7365
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7599..7616
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7733..7754
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7769..7774,7775..7792,7793..7799)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7807..7823
                     /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     7812..7834
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7831..7847
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7831..7847
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7866..7886
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7868..7886
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7914..8140
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8141..8321
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8368..8493
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8986..9219
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9404..9448
                     /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"
ORIGIN
        1 aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
       61 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
      121 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
      181 gagatccact ttggccgcga atcgatatgt cgagtttact ccctatcagt gatagagaac
      241 gtatgtcgag tttactccct atcagtgata gagaacgatg tcgagtttac tccctatcag
      301 tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt
      361 actccctatc agtgatagag aacgtatgtc gagtttatcc ctatcagtga tagagaacgt
      421 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg
      481 ggaggcctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gaattggcta
      541 gcgccaccat ggctgccatc cggaagaaac tggtgattgt tggtgatgta gcctgtggaa
      601 agacatgctt gctcatagtc ttcagcaagg accagttccc agaggtgtat gtgcccacag
      661 tgtttgagaa ctatgtggca gatatcgagg tggatggaaa gcaggtagag ttggctttgt
      721 gggacacagc tgggcaggaa gattatgatc gcctgaggcc cctctcctac ccagataccg
      781 atgttatact gatgtgtttt tccatcgaca gccctgatag tttagaaaac atcccagaaa
      841 agtggacccc agaagtcaag catttctgtc ccaacgtgcc catcatcctg gttgggaata
      901 agaaggatct tcggaatgat gagcacacaa ggcgggagct agccaagatg aagcaggagc
      961 cggtgaaacc tgaagaaggc agagatatgg caaacaggat tggcgctttt gggtacatgg
     1021 agtgttcagc aaagaccaaa gatggagtga gagaggtttt tgaaatggct acgagagctg
     1081 ctctgcaagc tagacgtggg aagaaaaaat ctgggtgcct tgtcttggga tacccatacg
     1141 atgttcctga ctatgcgggc tatccctatg acgtcccgga ctatgcaggg ccggctaagg
     1201 cagcggcacg ggcggtgtag ctcgagggcc ggcgcgccgc ggccgctacg taaattccgc
     1261 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     1321 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     1381 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     1441 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     1501 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     1561 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     1621 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     1681 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
     1741 atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     1801 cctttgaaaa acacgatgat aatatggcca caaccatggt gagcaagggc gaggagctgt
     1861 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     1921 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     1981 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     2041 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2101 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     2161 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     2221 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     2281 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     2341 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     2401 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     2461 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     2521 ggatcactct cggcatggac gagctgtaca agtaaggatc cggggttggg gttgcgcctt
     2581 ttccaaggca gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa
     2641 cgcagcggcg ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg
     2701 atcttcgccg ctacccttgt gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg
     2761 gaaggttcct tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact
     2821 agtaccctcg cagacggaca gcgccaggga gcaatggcag cgcgccgacc gcgatgggct
     2881 gtggccaata gcggctgctc agcagggcgc gccgagagca gcggccggga aggggcggtg
     2941 cgggaggcgg ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca
     3001 ttctgcaagc ctccggagcg cacgtcggca gtcggctccc tcgttgaccg aatcaccgac
     3061 ctctctcccc agcaattcac catgaccgag tacaagccca cggtgcgcct cgccacccgc
     3121 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     3181 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     3241 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     3301 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     3361 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     3421 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     3481 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     3541 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     3601 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     3661 acccgcaagc ccggtgccga aggtagaggt tctctcctca cttgtggtga tgttgaagaa
     3721 aaccctggtc caatgtctag actggacaag agcaaagtca taaacggagc tctggaatta
     3781 ctcaatggtg tcggtatcga aggcctgacg acaaggaaac tcgctcaaaa gctgggagtt
     3841 gagcagccta ccctgtactg gcacgtgaag aacaagcggg ccctgctcga tgccctgcca
     3901 atcgagatgc tggacaggca tcatacccac ttctgccccc tggaaggcga gtcatggcaa
     3961 gactttctgc ggaacaacgc caagtcatac cgctgtgctc tcctctcaca tcgcgacggg
     4021 gctaaagtgc atctcggcac ccgcccaaca gagaaacagt acgaaaccct ggaaaatcag
     4081 ctcgcgttcc tgtgtcagca aggcttctcc ctggagaacg cactgtacgc tctgtccgcc
     4141 gtgggccact ttacactggg ctgcgtattg gaggaacagg agcatcaagt agcaaaagag
     4201 gaaagagaga cacctaccac cgattctatg cccccacttc tgagacaagc aattgagctg
     4261 ttcgaccggc agggagccga acctgccttc cttttcggcc tggaactaat catatgtggc
     4321 ctggagaaac agctaaagtg cgaaagcggc gggccgaccg acgcccttga cgattttgac
     4381 ttagacatgc tcccagccga tgcccttgac gactttgacc ttgatatgct gcctgctgac
     4441 gctcttgacg attttgacct tgacatgctc cccgggtaac taagtaagga tcgatccaag
     4501 atatcgtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     4561 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     4621 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     4681 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     4741 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     4801 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     4861 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     4921 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     4981 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     5041 ggccgcctcc ccgcctgttt cgcctcggcg tccggactag aggtaccttt aagaccaatg
     5101 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     5161 ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta
     5221 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     5281 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     5341 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt
     5401 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac
     5461 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5521 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5581 catgtctggc tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5641 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5701 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctttcaa
     5761 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5821 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5881 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5941 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6001 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6061 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6121 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6181 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6241 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6301 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6361 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6421 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6481 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6541 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6601 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6661 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6721 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6781 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6841 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6901 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6961 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     7021 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7081 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7141 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7201 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7261 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7321 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7381 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7441 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7501 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     7561 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     7621 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     7681 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     7741 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     7801 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     7861 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc
     7921 ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca
     7981 aggagagaaa aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt
     8041 attaggaagg caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg
     8101 cagagatatt gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca
     8161 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     8221 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     8281 atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac
     8341 ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg
     8401 cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga
     8461 ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg
     8521 cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata
     8581 gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca
     8641 gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa
     8701 cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata
     8761 aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc
     8821 gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag
     8881 aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa
     8941 ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct
     9001 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca
     9061 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga
     9121 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat
     9181 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg
     9241 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga
     9301 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag
     9361 cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt
     9421 attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg
     9481 gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc
     9541 tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca
     9601 cctcccaacc ccgaggggac
//
