LOCUS       CD81__TEV__Cre_M        7232 bp ds-DNA     linear       01-MAR-2021
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"
     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"
     primer_bind     complement(2826..2843)
                     /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    2832..3056
                     /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      3102..3530
                     /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(3189..3208)
                     /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     3399..3420
                     /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(3539..3559)
                     /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        3544..3873
                     /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      3724..3859
                     /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     3786..3805
                     /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             3940..4734
                     /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(3994..4013)
                     /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     4604..4623
                     /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    4908..5029
                     /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(4945..4964)
                     /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     4999..5018
                     /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(5078..5094)
                     /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
     primer_bind     complement(5078..5094)
                     /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
     CDS             995..1699
                     /locus_tag="CD81"
                     /label="CD81"
                     /ApEinfo_label="CD81"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             1721..2788
                     /locus_tag="Cre-Myc"
                     /label="Cre-Myc"
                     /ApEinfo_label="Cre-Myc"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(5091..5113)
                     /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
     mat_peptide     2756..2785
                     /locus_tag="Myc tag"
                     /label="Myc tag"
                     /ApEinfo_label="Myc tag"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     protein_bind    5102..5118
                     /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(5126..5156)
                     /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    5171..5192
                     /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(5309..5326)
                     /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(5480..6065)
                     /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(5560..5579)
                     /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(6236..7096)
                     /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     6859..6878
                     /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(7097..7201)
                     /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             1700..1720
                     /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"
     primer_bind     complement(5074..5094)
                     /locus_tag="M13-rev"
                     /label="M13-rev"
                     /ApEinfo_label="M13-rev"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     863..882
                     /locus_tag="T7"
                     /label="T7"
                     /ApEinfo_label="T7"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     rep_origin      3119..3425
                     /locus_tag="F1 ori"
                     /label="F1 ori"
                     /ApEinfo_label="F1 ori"
                     /ApEinfo_fwdcolor="gray50"
                     /ApEinfo_revcolor="gray50"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_binding    complement(5100..5122)
                     /locus_tag="LacO"
                     /label="LacO"
                     /ApEinfo_label="LacO"
                     /ApEinfo_fwdcolor="#6495ed"
                     /ApEinfo_revcolor="#6495ed"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             complement(6239..6898)
                     /locus_tag="AmpR(1)"
                     /label="AmpR(1)"
                     /ApEinfo_label="AmpR"
                     /ApEinfo_fwdcolor="yellow"
                     /ApEinfo_revcolor="yellow"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             3943..4734
                     /locus_tag="Kan/neoR"
                     /label="Kan/neoR"
                     /ApEinfo_label="Kan/neoR"
                     /ApEinfo_fwdcolor="yellow"
                     /ApEinfo_revcolor="yellow"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_binding    1571..1576
                     /locus_tag="Cla I"
                     /label="Cla I"
                     /ApEinfo_label="Cla I"
                     /ApEinfo_fwdcolor="#ff4809"
                     /ApEinfo_revcolor="#ff4809"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_binding    2523..2528
                     /locus_tag="Cla I(1)"
                     /label="Cla I(1)"
                     /ApEinfo_label="Cla I"
                     /ApEinfo_fwdcolor="#ff4809"
                     /ApEinfo_revcolor="#ff4809"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     6859..6878
                     /locus_tag="Amp-R (& -comp-R)"
                     /label="Amp-R (& -comp-R)"
                     /ApEinfo_label="Amp-R (& -comp-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(4945..4964)
                     /locus_tag="SV40-pA-R"
                     /label="SV40-pA-R"
                     /ApEinfo_label="SV40-pA-R"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     3786..3805
                     /locus_tag="SV40-Pro"
                     /label="SV40-Pro"
                     /ApEinfo_label="SV40-Pro"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     1651..1670
                     /locus_tag="CD81-3'"
                     /label="CD81-3'"
                     /ApEinfo_label="CD81-3'"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /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 catgggagtg gagggctgca ccaagtgcat
     1021 caagtacctg ctcttcgtct tcaatttcgt cttctggctg gctggaggcg tgatcctggg
     1081 tgtggccctg tggctccgcc atgacccgca gaccaccaac ctcctgtatc tggagctggg
     1141 agacaagccc gcgcccaaca ccttctatgt aggcatctac atcctcatcg ctgtgggcgc
     1201 tgtcatgatg ttcgttggct tcctgggctg ctacggggcc atccaggaat cccagtgcct
     1261 gctggggacg ttcttcacct gcctggtcat cctgtttgcc tgtgaggtgg ccgccggcat
     1321 ctggggcttt gtcaacaagg accagatcgc caaggatgtg aagcagttct atgaccaggc
     1381 cctacagcag gccgtggtgg atgatgacgc caacaacgcc aaggctgtgg tgaagacctt
     1441 ccacgagacg cttgactgct gtggctccag cacactgact gctttgacca cctcagtgct
     1501 caagaacaat ttgtgtccct cgggcagcaa catcatcagc aacctcttca aggaggactg
     1561 ccaccagaag atcgatgacc tcttctccgg gaagctgtac ctcatcggca ttgctgccat
     1621 cgtggtcgct gtgatcatga tcttcgagat gatcctgagc atggtgctgt gctgtggcat
     1681 ccggaacagc tccgtgtacg agaacctgta cttccagtct GCCAATTTAC TGACCGTACA
     1741 CCAAAATTTG CCTGCATTAC CGGTCGATGC AACGAGTGAT GAGGTTCGCA AGAACCTGAT
     1801 GGACATGTTC AGGGATCGCC AGGCGTTTTC TGAGCATACC TGGAAAATGC TTCTGTCCGT
     1861 TTGCCGGTCG TGGGCGGCAT GGTGCAAGTT GAATAACCGG AAATGGTTTC CCGCAGAACC
     1921 TGAAGATGTT CGCGATTATC TTCTATATCT TCAGGCGCGC GGTCTGGCAG TAAAAACTAT
     1981 CCAGCAACAT TTGGGCCAGC TAAACATGCT TCATCGTCGG TCCGGGCTGC CACGACCAAG
     2041 TGACAGCAAT GCTGTTTCAC TGGTTATGCG GCGGATCCGA AAAGAAAACG TTGATGCCGG
     2101 TGAACGTGCA AAACAGGCTC TAGCGTTCGA ACGCACTGAT TTCGACCAGG TTCGTTCACT
     2161 CATGGAAAAT AGCGATCGCT GCCAGGATAT ACGTAATCTG GCATTTCTGG GGATTGCTTA
     2221 TAACACCCTG TTACGTATAG CCGAAATTGC CAGGATCAGG GTTAAAGATA TCTCACGTAC
     2281 TGACGGTGGG AGAATGTTAA TCCATATTGG CAGAACGAAA ACGCTGGTTA GCACCGCAGG
     2341 TGTAGAGAAG GCACTTAGCC TGGGGGTAAC TAAACTGGTC GAGCGATGGA TTTCCGTCTC
     2401 TGGTGTAGCT GATGATCCGA ATAACTACCT GTTTTGCCGG GTCAGAAAAA ATGGTGTTGC
     2461 CGCGCCATCT GCCACCAGCC AGCTATCAAC TCGCGCCCTG GAAGGGATTT TTGAAGCAAC
     2521 TCATCGATTG ATTTACGGCG CTAAGGATGA CTCTGGTCAG AGATACCTGG CCTGGTCTGG
     2581 ACACAGTGCC CGTGTCGGAG CCGCGCGAGA TATGGCCCGC GCTGGAGTTT CAATACCGGA
     2641 GATCATGCAA GCTGGTGGCT GGACCAATGT AAATATTGTC ATGAACTATA TCCGTAACCT
     2701 GGATAGTGAA ACAGGGGCAA TGGTGCGCCT GCTGGAAGAT GGCGATGGAC CGGTGGAACA
     2761 AAAACTTATT TCTGAAGAAG ATCTGTGAct cgagtctaga gggcccgttt aaacccgctg
     2821 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2881 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2941 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3001 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3061 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3361 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3421 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3481 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     3541 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3601 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3661 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3721 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3781 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     3841 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     3901 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     3961 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4021 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4081 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4141 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4201 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4261 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4321 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4381 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4441 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4501 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4561 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4621 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4681 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     4741 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     4801 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     4861 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     4921 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4981 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     5041 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5101 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5161 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5221 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5281 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5341 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5401 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5461 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5521 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5581 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5641 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5701 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5761 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5821 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5881 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5941 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6001 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6061 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6121 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6181 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6241 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6301 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6361 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6421 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6481 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6541 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6601 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6661 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6721 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6781 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6841 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6901 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6961 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7021 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7081 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7141 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7201 gcacatttcc ccgaaaagtg ccacctgacg tc  
//
