LOCUS       CD9_TEV_tTA             7142 bp ds-DNA     linear       26-OCT-2020
DEFINITION  Expression of G protein-coupled receptors for PRESTO-Tango:
            parallel receptorome expression and screening via transcriptional
            output,  with transcriptional activation following arrestin
            translocation.
ACCESSION   .
VERSION     .
KEYWORDS    ADRB2-Tango
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6632)
  AUTHORS   Kroeze WK, Sassano MF, Huang XP, Lansu K, McCorvy JD, Giguere PM,
            Sciaky N, Roth BL
  TITLE     PRESTO-Tango as an open-source resource for interrogation of the
            druggable human GPCRome.
  JOURNAL   Nat Struct Mol Biol. 2015 May;22(5):362-9. doi: 10.1038/nsmb.3014.
            Epub 2015 Apr 20.
  PUBMED    25895059
REFERENCE   2  (bases 1 to 6632)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 8, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
COMMENT     
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     primer_bind     complement(44..63)
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
                     /locus_tag="pRS-marker"
                     /label="pRS-marker"
                     /ApEinfo_label="pRS-marker"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /locus_tag="CMV enhancer"
                     /label="CMV enhancer"
                     /ApEinfo_label="CMV enhancer"
                     /ApEinfo_fwdcolor="#5ac3fa"
                     /ApEinfo_revcolor="#5ac3fa"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /locus_tag="CMV promoter"
                     /label="CMV promoter"
                     /ApEinfo_label="CMV promoter"
                     /ApEinfo_fwdcolor="#346ee0"
                     /ApEinfo_revcolor="#346ee0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     769..789
                     /note="Human CMV immediate early promoter, forward primer"
                     /locus_tag="CMV-F"
                     /label="CMV-F"
                     /ApEinfo_label="CMV-F"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     863..882
                     /note="T7 promoter, forward primer"
                     /locus_tag="T7"
                     /label="T7"
                     /ApEinfo_label="T7"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /locus_tag="T7 promoter"
                     /label="T7 promoter"
                     /ApEinfo_label="T7 promoter"
                     /ApEinfo_fwdcolor="#346ee0"
                     /ApEinfo_revcolor="#346ee0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             1697..2698
                     /codon_start=1
                     /product="tetracycline-controlled transactivator,
                     comprising a fusion of the tetracycline repressor TetR
                     with the C-terminal activation domain of herpes simplex
                     virus  VP16"
                     /note="In the Tet-Off(R) system, tTA binds to the
                     Tet-responsive element and stimulates transcription only
                     in the absence of tetracycline or doxycycline."
                     /translation="RLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVK
                     N
                     KRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRP
                     T
                     EKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTD
                     S
                     MPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTIE
                     G
                     LLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHAD
                     A LDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
                     /locus_tag="tTA"
                     /label="tTA"
                     /ApEinfo_label="tTA"
                     /ApEinfo_fwdcolor="#5518fe"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(1764..1783)
                     /note="Tetracycline resistance gene, reverse primer"
                     /locus_tag="Tet-R"
                     /label="Tet-R"
                     /ApEinfo_label="Tet-R"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(2736..2753)
                     /note="Bovine growth hormone terminator, reverse primer.
                     Also called BGH reverse"
                     /locus_tag="BGH-rev"
                     /label="BGH-rev"
                     /ApEinfo_label="BGH-rev"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     polyA_signal    2742..2966
                     /note="bovine growth hormone polyadenylation signal"
                     /locus_tag="bGH poly(A) signal"
                     /label="bGH poly(A) signal"
                     /ApEinfo_label="bGH poly(A) signal"
                     /ApEinfo_fwdcolor="#ff3eee"
                     /ApEinfo_revcolor="#ff3eee"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     rep_origin      3012..3440
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /locus_tag="f1 ori"
                     /label="f1 ori"
                     /ApEinfo_label="f1 ori"
                     /ApEinfo_fwdcolor="#999999"
                     /ApEinfo_revcolor="#999999"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     primer_bind     complement(3099..3118)
                     /note="F1 origin, reverse primer"
                     /locus_tag="F1ori-R"
                     /label="F1ori-R"
                     /ApEinfo_label="F1ori-R"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     primer_bind     3309..3330
                     /note="F1 origin, forward primer"
                     /locus_tag="F1ori-F"
                     /label="F1ori-F"
                     /ApEinfo_label="F1ori-F"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     primer_bind     complement(3449..3469)
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
                     /locus_tag="pBABE 3'"
                     /label="pBABE 3'"
                     /ApEinfo_label="pBABE 3'"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     promoter        3454..3783
                     /note="SV40 enhancer and early promoter"
                     /locus_tag="SV40 promoter"
                     /label="SV40 promoter"
                     /ApEinfo_label="SV40 promoter"
                     /ApEinfo_fwdcolor="#346ee0"
                     /ApEinfo_revcolor="#346ee0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     rep_origin      3634..3769
                     /note="SV40 origin of replication"
                     /locus_tag="SV40 ori"
                     /label="SV40 ori"
                     /ApEinfo_label="SV40 ori"
                     /ApEinfo_fwdcolor="#999999"
                     /ApEinfo_revcolor="#999999"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     3696..3715
                     /note="SV40 promoter/origin, forward primer"
                     /locus_tag="SV40pro-F"
                     /label="SV40pro-F"
                     /ApEinfo_label="SV40pro-F"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             3850..4644
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     P
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     S
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     Q
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     A LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
                     /locus_tag="NeoR/KanR"
                     /label="NeoR/KanR"
                     /ApEinfo_label="NeoR/KanR"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(3904..3923)
                     /note="Neomycin resistance gene, reverse primer"
                     /locus_tag="Neo-R"
                     /label="Neo-R"
                     /ApEinfo_label="Neo-R"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     primer_bind     4514..4533
                     /note="Neomycin resistance gene, forward primer"
                     /locus_tag="Neo-F"
                     /label="Neo-F"
                     /ApEinfo_label="Neo-F"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     polyA_signal    4818..4939
                     /note="SV40 polyadenylation signal"
                     /locus_tag="SV40 poly(A) signal"
                     /label="SV40 poly(A) signal"
                     /ApEinfo_label="SV40 poly(A) signal"
                     /ApEinfo_fwdcolor="#ff3eee"
                     /ApEinfo_revcolor="#ff3eee"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(4855..4874)
                     /note="SV40 polyA, reverse primer"
                     /locus_tag="SV40pA-R"
                     /label="SV40pA-R"
                     /ApEinfo_label="SV40pA-R"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     4909..4928
                     /note="SV40 polyA terminator, reverse primer"
                     /locus_tag="EBV-rev"
                     /label="EBV-rev"
                     /ApEinfo_label="EBV-rev"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     primer_bind     complement(4988..5004)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /locus_tag="M13 rev"
                     /label="M13 rev"
                     /ApEinfo_label="M13 rev"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     CDS             995..1675
                     /locus_tag="CD9"
                     /label="CD9"
                     /ApEinfo_label="CD9"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(4988..5004)
                     /note="In lacZ gene. Also called M13-rev"
                     /locus_tag="M13 Reverse"
                     /label="M13 Reverse"
                     /ApEinfo_label="M13 Reverse"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     primer_bind     complement(5001..5023)
                     /note="In lacZ gene"
                     /locus_tag="M13/pUC Reverse"
                     /label="M13/pUC Reverse"
                     /ApEinfo_label="M13/pUC Reverse"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     protein_bind    5012..5028
                     /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)."
                     /locus_tag="lac operator"
                     /label="lac operator"
                     /ApEinfo_label="lac operator"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     promoter        complement(5036..5066)
                     /note="promoter for the E. coli lac operon"
                     /locus_tag="lac promoter"
                     /label="lac promoter"
                     /ApEinfo_label="lac promoter"
                     /ApEinfo_fwdcolor="#346ee0"
                     /ApEinfo_revcolor="#346ee0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     protein_bind    5081..5102
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /locus_tag="CAP binding site"
                     /label="CAP binding site"
                     /ApEinfo_label="CAP binding site"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     primer_bind     complement(5219..5236)
                     /note="L4440 vector, forward primer"
                     /locus_tag="L4440"
                     /label="L4440"
                     /ApEinfo_label="L4440"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     rep_origin      complement(5390..5975)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /locus_tag="ori"
                     /label="ori"
                     /ApEinfo_label="ori"
                     /ApEinfo_fwdcolor="#999999"
                     /ApEinfo_revcolor="#999999"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     primer_bind     complement(5470..5489)
                     /note="pBR322 origin, forward primer"
                     /locus_tag="pBR322ori-F"
                     /label="pBR322ori-F"
                     /ApEinfo_label="pBR322ori-F"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     CDS             complement(6146..7006)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     I
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     S
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     W
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     A
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     S LIKHW"
                     /locus_tag="AmpR"
                     /label="AmpR"
                     /ApEinfo_label="AmpR"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     6769..6788
                     /note="Ampicillin resistance gene, reverse primer"
                     /locus_tag="Amp-R"
                     /label="Amp-R"
                     /ApEinfo_label="Amp-R"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     promoter        complement(7007..7111)
                     /gene="bla"
                     /locus_tag="AmpR promoter"
                     /label="AmpR promoter"
                     /ApEinfo_label="AmpR promoter"
                     /ApEinfo_fwdcolor="#346ee0"
                     /ApEinfo_revcolor="#346ee0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     CDS             1676..1696
                     /locus_tag="TEV protease cleavage site"
                     /label="TEV protease cleavage site"
                     /ApEinfo_label="TEV protease cleavage site"
                     /ApEinfo_fwdcolor="#fb5261"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc
      961 agatatccag cacagtggcg gccgcgccac catgccggtc aaaggaggca ccaagtgcat
     1021 caaatacctg ctgttcggat ttaacttcat cttctggctt gccgggattg ctgtccttgc
     1081 cattggacta tggctccgat tcgactctca gaccaagagc atcttcgagc aagaaactaa
     1141 taataataat tccagcttct acacaggagt ctatattctg atcggagccg gcgccctcat
     1201 gatgctggtg ggcttcctgg gctgctgcgg ggctgtgcag gagtcccagt gcatgctggg
     1261 actgttcttc ggcttcctct tggtgatatt cgccattgaa atagctgcgg ccatctgggg
     1321 atattcccac aaggatgagg tgattaagga agtccaggag ttttacaagg acacctacaa
     1381 caagctgaaa accaaggatg agccccagcg ggaaacgctg aaagccatcc actatgcgtt
     1441 gaactgctgt ggtttggctg ggggcgtgga acagtttatc tcagacatct gccccaagaa
     1501 ggacgtactc gaaaccttca ccgtgaagtc ctgtcctgat gccatcaaag aggtcttcga
     1561 caataaattc cacatcatcg gcgcagtggg catcggcatt gccgtggtca tgatatttgg
     1621 catgatcttc agtatgatct tgtgctgtgc tatccgcagg aaccgcgaga tggtcgagaa
     1681 cctgtacttc cagtctagat tagataaaag taaagtgatt aacagcgcat tagagctgct
     1741 taatgaggtc ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc taggtgtaga
     1801 gcagcctaca ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg ccttagccat
     1861 tgagatgtta gataggcacc atactcactt ttgcccttta gaaggggaaa gctggcaaga
     1921 ttttttacgt aataacgcta aaagttttag atgtgcttta ctaagtcatc gcgatggagc
     1981 aaaagtacat ttaggtacac ggcctacaga aaaacagtat gaaactctcg aaaatcaatt
     2041 agccttttta tgccaacaag gtttttcact agagaatgca ttatatgcac tcagcgctgt
     2101 ggggcatttt actttaggtt gcgtattgga agatcaagag catcaagtcg ctaaagaaga
     2161 aagggaaaca cctactactg atagtatgcc gccattatta cgacaagcta tcgaattatt
     2221 tgatcaccaa ggtgcagagc cagccttctt attcggcctt gaattgatca tatgcggatt
     2281 agaaaaacaa cttaaatgtg aaagtgggtc cgcgtacagc cgcgcgcgta cgaaaaacaa
     2341 ttacgggtct accatcgagg gcctgctcga tctcccggac gacgacgccc ccgaagaggc
     2401 ggggctggcg gctccgcgcc tgtcctttct ccccgcggga cacacgcgca gactgtcgac
     2461 ggcccccccg accgatgtca gcctggggga cgagctccac ttagacggcg aggacgtggc
     2521 gatggcgcat gccgacgcgc tagacgattt cgatctggac atgttggggg acggggattc
     2581 cccgggtccg ggatttaccc cccacgactc cgccccctac ggcgctctgg atatggccga
     2641 cttcgagttt gagcagatgt ttaccgatgc ccttggaatt gacgagtacg gtgggtagct
     2701 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     2761 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2821 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2881 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     2941 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3001 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3061 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3421 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3481 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3541 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3601 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3661 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3721 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3781 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     3841 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3901 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3961 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4021 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4081 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4141 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4201 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4261 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4321 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4381 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4441 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4501 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4561 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4621 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4681 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4741 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4801 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4861 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4921 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4981 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5041 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5101 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5161 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5221 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5281 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5341 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5401 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5461 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5521 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5581 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5641 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5701 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5761 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5821 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5881 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     5941 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6001 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6061 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6121 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6181 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6241 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6301 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6361 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6421 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6481 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6541 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6601 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6661 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6721 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6781 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6841 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6901 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6961 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7021 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7081 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7141 tc     
//
