LOCUS       FSW_SUN1_GFP       10833 bp DNA     circular SYN 28-JUN-2023
DEFINITION  synthetic DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10833)
  TITLE     NEBuilder-generated Construct
  JOURNAL   Exported  5-JUN-2023 from NEBuilder https://nebuilder.neb.com
REFERENCE   2  (bases 1 to 10833)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 12, 2023 from SnapGene 7.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Exported  
            5-JUN-2023 from NEBuilder https://nebuilder.neb.com"
COMMENT     NEBuilder-generated oligos (UPPERCASE = gene-specific, lowercase = 
            overlap)
            Sun1-GFP_fwd: cgacgtcggttggcgcgccgccaccatggccACCTCCAGGGTGTACTCC
            Sun1-GFP_fwd 3'Tm: 65.9  3'Ta: 63.1
            Sun1-GFP_rev: gataaacccttggcgcgccgTTATTTGTAGAGCTCATCCATGC
            Sun1-GFP_rev 3'Tm: 62.1  3'Ta: 63.1
FEATURES             Location/Qualifiers
     source          1..10833
                     /plasmid="FSW_SUN_GFP"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
                     /organism="synthetic DNA construct"
     gene            1..8455
                     /label=FSW-tdTomato
                     /note="FSW-tdTomato"
                     /note="color: #a6acb3; direction: RIGHT"
     primer_bind     complement(9..46)
                     /label=hSyn1-tdTomato_rev
                     /note="hSyn1-tdTomato_rev"
                     /note="gene-specific Tm: 56.2  Ta: 57.2"
                     /note="overlap: tccagaggttgattatcgat"
                     /note="gene-specific primer: AAGCTTGATAAACCCTTG"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(9..46)
                     /label=hSyn1_tdTomato_R
                     /note="color: black; sequence: 
                     tccagaggttgattatcgataagcttgataaacccttg; added: 2023-06-06"
     primer_bind     complement(20..46)
                     /label=tdTom_R
                     /note="color: black; sequence: tccagaggttgattatcgataagcttg;
                     added: 2023-06-06"
     primer_bind     21..58
                     /label=FSW_Hin_F
                     /note="color: black; sequence: 
                     atttatggtattgttgtagaaagcttatcgataatcaacctctggattacaaaatttg;
                      added: 2023-06-06"
     gene            27..7963
                     /label=FSW-H3MD-Vector
                     /note="FSW-H3MD-Vector"
                     /note="color: #a6acb3; direction: RIGHT"
     primer_bind     27..58
                     /label=FSW-H3MD-Vector_fwd
                     /note="FSW-H3MD-Vector_fwd"
                     /note="gene-specific Tm: 64.7  Ta: 65.7"
                     /note="gene-specific primer: 
                     ATCGATAATCAACCTCTGGATTACAAAATTTG"
                     /note="color: #a020f0; direction: RIGHT"
     misc_feature    33..621
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     primer_bind     35..60
                     /label=FSW_Tom_F
                     /note="color: black; sequence: tcaacctctggattacaaaatttgtg; 
                     added: 2023-06-06"
     CDS             complement(504..515)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 503"
                     /translation="IEGR"
     primer_bind     complement(624..640)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     625..644
                     /label=LVseq3F
                     /note="color: black; sequence: GATACCGTCGACCTCGAGAC; added:
                     2023-06-06"
     primer_bind     complement(625..641)
                     /label=pBluescript KS
                     /note="Stratagene"
                     /note="color: black; sequence: tcgaggtcgacggtatc; added: 
                     2023-06-06"
     primer_bind     911..930
                     /label=HIVscreenF
                     /note="color: black; sequence: GAACTACACACCAGGGCCAG; added:
                     2023-06-06"
     primer_bind     911..930
                     /label=LTRdet_F1
                     /note="color: black; sequence: GAACTACACACCAGGGCCAG; added:
                     2023-06-06"
     LTR             1146..1326
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /note="color: #ffe4c4"
     primer_bind     complement(1196..1215)
                     /label=LTRdet_R
                     /note="color: black; sequence: GCTTAAGCAGTGGGTTCCCT; added:
                     2023-06-06"
     polyA_signal    1358..1582
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     polyA_signal    1358..1565
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     1462..1481
                     /label=BGHseq_F
                     /note="color: black; sequence: ATTGCATCGCATTGTCTGAG; added:
                     2023-06-06"
     primer_bind     complement(1574..1593)
                     /label=Puro-R
                     /note="color: black; sequence: CGCCTCAGAAGCCATAGAGC; added:
                     2023-06-06"
     rep_origin      1628..2056
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     primer_bind     complement(1715..1734)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
                     /note="color: black; sequence: AGGGAAGAAAGCGAAAGGAG; added:
                     2023-06-06"
     primer_bind     1925..1946
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
                     /note="color: black; sequence: GTGGACTCTTGTTCCAAACTGG; 
                     added: 2023-06-06"
     promoter        2070..2399
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      2250..2385
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     promoter        2447..2494
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
                     /note="color: #ffffff; direction: RIGHT"
     CDS             2513..2887
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /note="color: #ccffcc"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3017..3138
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(3187..3203)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(3200..3222)
                     /label=M13F-48
                     /note="color: black; sequence: AGCGGATAACAATTTCACACAGG; 
                     added: 2023-06-06"
     protein_bind    3211..3227
                     /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)."
                     /note="color: #31849b"
     primer_bind     complement(3228..3247)
                     /label=pSC_F
                     /note="color: black; sequence: GGCTCGTATGTTGTGTGGAA; added:
                     2023-06-06"
     promoter        complement(3235..3265)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 3235 .. 3241 / #ffffff / -10
                        2: 3242 .. 3259 / #ffffff
                        3: 3260 .. 3265 / #ffffff / -35"
     primer_bind     3262..3281
                     /label=PuroBleo_seq_F
                     /note="color: black; sequence: TAAAGCCTGGGGTGCCTAAT; added:
                     2023-06-06"
     protein_bind    3280..3301
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     primer_bind     complement(3473..3516)
                     /label=PuroBleo_R
                     /note="color: black; sequence: 
                     TTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTG; added: 
                     2023-06-06"
     primer_bind     3492..3509
                     /label=partTetVec_F
                     /note="color: black; sequence: GGTTATCCACAGAATCAG; added: 
                     2023-06-06"
     primer_bind     3497..3521
                     /label=partTetVec2L_F
                     /note="color: black; sequence: TCCACAGAATCAGGGGATAACGCAG; 
                     added: 2023-06-06"
     primer_bind     3497..3517
                     /label=partTetVec2_fwd
                     /note="color: black; sequence: TCCACAGAATCAGGGGATAAC; 
                     added: 2023-06-06"
     rep_origin      complement(3589..4177)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     complement(3669..3688)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT; added:
                     2023-06-06"
     CDS             complement(4348..5208)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 4348 .. 5139 / #ccffcc
                        2: 5140 .. 5208 / #ccffcc / signal sequence
                     Cleavage site after base 5139"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4600..4619
                     /label=AmpR-F
                     /note="color: black; sequence: TTGCCGGGAAGCTAGAGTAA; added:
                     2023-06-06"
     primer_bind     4971..4990
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC; added:
                     2023-06-06"
     primer_bind     complement(4971..4990)
                     /label=AmpR-R
                     /note="color: black; sequence: GCTATGTGGCGCGGTATTAT; added:
                     2023-06-06"
     promoter        complement(5209..5313)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     5243..5265
                     /label=OS280
                     /note="color: black; sequence: CAGGGTTATTGTCTCATGAGCGG; 
                     added: 2023-06-06"
     primer_bind     complement(5388..5407)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
                     /note="color: black; sequence: CGGCATCAGAGCAGATTGTA; added:
                     2023-06-06"
     enhancer        5579..5958
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     primer_bind     complement(5825..5844)
                     /label=CAGseq_R
                     /note="color: black; sequence: GGGCGTACTTGGCATATGAT; added:
                     2023-06-06"
     promoter        5960..6158
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     6061..6080
                     /label=CMVenh_F
                     /note="color: black; sequence: CACCAAAATCAACGGGACTT; added:
                     2023-06-06"
     primer_bind     6075..6092
                     /label=VP1.5
                     /note="color: black; sequence: GGACTTTCCAAAATGTCG; added: 
                     2023-06-06"
     LTR             6176..6356
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /note="color: #ffe4c4"
     primer_bind     complement(6226..6245)
                     /label=LTRdet_R
                     /note="color: black; sequence: GCTTAAGCAGTGGGTTCCCT; added:
                     2023-06-06"
     misc_feature    6403..6528
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
                     /note="color: #ffe4c4"
     primer_bind     complement(6460..6480)
                     /label=LVseq1R
                     /note="color: black; sequence: TTTTGGCGTACTCACCAGTCG; 
                     added: 2023-06-06"
     primer_bind     6760..6780
                     /label=LVseq1F
                     /note="color: black; sequence: GCAACCCTCTATTGTGTGCAT; 
                     added: 2023-06-06"
     misc_feature    7021..7254
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
                     /note="color: #ffe4c4"
     primer_bind     complement(7212..7231)
                     /label=LVseq2R
                     /note="color: black; sequence: TCTTTCCACAGCCAGGATTC; added:
                     2023-06-06"
     CDS             7439..7483
                     /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"
                     /note="color: #cc99b2"
                     /translation="KNEQELLELDKWASL"
     primer_bind     7497..7518
                     /label=LVseq2F
                     /note="color: black; sequence: ACATAACAAATTGGCTGTGGTA; 
                     added: 2023-06-06"
     primer_bind     7661..7680
                     /label=RRE_F
                     /note="color: black; sequence: AGGCCCGAAGGAATAGAAGA; added:
                     2023-06-06"
     misc_feature    7781..7898
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
                     /note="color: #ffe4c4"
     primer_bind     7807..7826
                     /label=cPPT_F
                     /note="color: black; sequence: GGTACAGTGCAGGGGAAAGA; added:
                     2023-06-06"
     primer_bind     7944..7981
                     /label=hSyn1-tdTomato_fwd
                     /note="hSyn1-tdTomato_fwd"
                     /note="gene-specific Tm: 59.3  Ta: 57.2"
                     /note="overlap: tgcagagggccctgcgtatg"
                     /note="gene-specific primer: AGTGCAAGTGGGTTTTAG"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     7944..7958
                     /label=hSynPro_F
                     /note="color: black; sequence: tgcagagggccctgc; added: 
                     2023-06-06"
     primer_bind     complement(7946..7963)
                     /label=FSW-H3MD-Vector_rev
                     /note="FSW-H3MD-Vector_rev"
                     /note="gene-specific Tm: 68.6  Ta: 65.7"
                     /note="gene-specific primer: CATACGCAGGGCCCTCTG"
                     /note="color: #a020f0; direction: LEFT"
     promoter        7964..8411
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     7964..7984
                     /label=tdTom_F
                     /note="color: black; sequence: agtgcaagtgggttttaggac; 
                     added: 2023-06-06"
     primer_bind     8088..8105
                     /label=hSynSeq_F
                     /note="color: black; sequence: gggaggggaaacaggatg; added: 
                     2023-06-06"
     primer_bind     complement(8123..8140)
                     /label=hSynSeq_R
                     /note="color: black; sequence: ggtgctgaagctggcagt; added: 
                     2023-06-06"
     primer_bind     complement(8394..8411)
                     /label=FSW_Nhe_R
                     /note="color: black; sequence: 
                     ccatggtggcgggatccttgctgcgctctcaggcacga; added: 2023-06-06"
     primer_bind     complement(8396..8411)
                     /label=FSW_R
                     /note="color: black; sequence: ctgcgctctcaggcac; added: 
                     2023-06-06"
     primer_bind     8436..8484
                     /label=Sun1-GFP_fwd
                     /note="Sun1-GFP_fwd"
                     /note="gene-specific Tm: 65.9  Ta: 63.1"
                     /note="overlap: cgacgtcggttggcgcgccg"
                     /note="spacer: ccaccatggcc"
                     /note="gene-specific primer: ACCTCCAGGGTGTACTCC"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     8436..8484
                     /label=Sun1_GFP_F
                     /note="color: black; sequence: 
                     cgacgtcggttggcgcgccgccaccatggccacctccagggtgtactcc; added: 
                     2023-06-05"
     regulatory      8455..8464
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     gene            8456..10833
                     /label=Sun1-GFP
                     /note="Sun1-GFP"
                     /note="color: #a6acb3; direction: RIGHT"
     CDS             8461..10833
                     /codon_start=1
                     /label=Sun1-GFP
                     /note="This feature has 3 segments:
                        1:  8461 ..  8466 / #999999 / Init
                        2:  8467 .. 10116 / #993366 / Sun1
                        3: 10117 .. 10833 / #00ff00 / Superfolder GFP"
                     /translation="MATSRVYSRDRTLKPRKAASGTFWWLGSGWYQFVTLISWLNVFLL
                     TRCLRNICKVFVLLLPLLLLLGAGVSLWGQGNFFSLLPVLNWTAMQPTQRVDDSKGMHR
                     PGPLPPSPPPKVDHKASQWPQESDMGQKVASLSAQCHNHDERLAELTVLLQKLQIRVDQ
                     VDDGREGLSLWVKNVVGQHLQEMGTIEPPDAKTDFMTFHHDHEVRLSNLEDVLRKLTEK
                     SEAIQKELEETKLKAGSRDEEQPLLDRVQHLELELNLLKSQLSDWQHLKTSCEQAGARI
                     QETVQLMFSEDQQGGSLEWLLEKLSSRFVSKDELQVLLHDLELKLLQNITHHITVTGQA
                     PTSEAIVSAVNQAGISGITEAQAHIIVNNALKLYSQDKTGMVDFALESGGGSILSTRCS
                     ETYETKTALLSLFGVPLWYFSQSPRVVIQPDIYPGNCWAFKGSQGYLVVRLSMKIYPTT
                     FTMEHIPKTLSPTGNISSAPKDFAVYGLETEYQEEGQPLGRFTYDQEGDSLQMFHTLER
                     PDQAFQIVELRVLSNWGHPEYTCLYRFRVHGEPIQMSKGEELFTGVVPILVELDGDVNG
                     HKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDF
                     FKSAMPEGYVQERTISFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLE
                     YNFNSHNVYITADKQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYL
                     STQSVLSKDPNEKRDHMVLLEFVTAAGITHGMDELYK"
     primer_bind     complement(8655..8674)
                     /label=Sun8674R
                     /note="color: black; sequence: gggagacaccagcacctagt; added:
                     2023-06-06"
     CDS             10117..10830
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="color: #05fd14"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     10421..10440
                     /label=SupGFP_F
                     /note="color: black; sequence: atgacgggacctacaagacg; added:
                     2023-06-06"
     primer_bind     complement(join(10811..10833,1..20))
                     /label=Sun1-GFP_rev
                     /note="Sun1-GFP_rev"
                     /note="gene-specific Tm: 62.1  Ta: 63.1"
                     /note="overlap: gataaacccttggcgcgccg"
                     /note="gene-specific primer: TTATTTGTAGAGCTCATCCATGC"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(join(10811..10833,1..20))
                     /label=Sun1_GFP_R
                     /note="color: black; sequence: 
                     gataaacccttggcgcgccgttatttgtagagctcatccatgc; added: 
                     2023-06-05"
ORIGIN
        1 cggcgcgcca agggtttatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
       61 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
      121 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
      181 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
      241 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
      301 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
      361 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
      421 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
      481 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
      541 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
      601 ccctttgggc cgcctccccg catcgatacc gtcgacctcg agacctagaa aaacatggag
      661 caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag
      721 aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca
      781 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
      841 actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc
      901 ctgattggca gaactacaca ccagggccag ggatcagata tccactgacc tttggatggt
      961 gctacaagct agtaccagtt gagcaagaga aggtagaaga agccaatgaa ggagagaaca
     1021 cccgcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtattag
     1081 agtggaggtt tgacagccgc ctagcatttc atcacatggc ccgagagctg catccggact
     1141 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     1201 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     1261 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     1321 ctagcagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     1381 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     1441 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     1501 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     1561 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     1621 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     1681 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     1741 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     1801 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     1861 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     1921 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     1981 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     2041 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     2101 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     2161 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2221 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2281 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     2341 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     2401 agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc
     2461 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa
     2521 gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg
     2581 gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg
     2641 ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct
     2701 ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc
     2761 cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg
     2821 gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca
     2881 ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     2941 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     3001 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3061 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3121 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     3181 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     3241 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     3301 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     3361 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3421 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3481 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3541 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3601 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3661 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3721 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3781 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3841 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3901 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3961 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4021 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4081 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4141 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4201 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4261 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4321 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4381 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4441 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4501 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4561 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4621 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4681 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4741 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4801 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4861 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4921 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4981 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5041 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5101 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5161 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5221 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5281 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     5341 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     5401 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     5461 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     5521 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
     5581 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     5641 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     5701 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     5761 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     5821 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     5881 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     5941 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     6001 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     6061 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     6121 ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg cgttttgcct gtactgggtc
     6181 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     6241 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     6301 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
     6361 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
     6421 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
     6481 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
     6541 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
     6601 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
     6661 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
     6721 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
     6781 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
     6841 aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat
     6901 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     6961 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     7021 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     7081 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     7141 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     7201 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     7261 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     7321 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     7381 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     7441 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     7501 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     7561 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     7621 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
     7681 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg
     7741 tgcgccaatt ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
     7801 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     7861 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
     7921 gagatccagt ttggttaatt aactgcagag ggccctgcgt atgagtgcaa gtgggtttta
     7981 ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca
     8041 agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca
     8101 ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc
     8161 cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg
     8221 gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag
     8281 ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg
     8341 gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc
     8401 tgagagcgca gacgcgtgga cggtaccctt caggtcgacg tcggttggcg cgccgccacc
     8461 atggccacct ccagggtgta ctccagagac aggactctca aaccacggaa ggcagcctcg
     8521 ggaaccttct ggtggctagg gagcggctgg taccaatttg ttactttgat ttcttggctg
     8581 aatgtctttc ttcttaccag gtgccttcga aatatttgca aggtttttgt cttgctcctc
     8641 ccactcctac ttttactagg tgctggtgtc tccctgtggg gccagggaaa cttcttctca
     8701 ctcctaccag tgctgaactg gacggccatg cagccaacac agagggtgga cgattccaag
     8761 ggcatgcata gacctggccc tcttcccccg agcccacctc caaaggttga tcacaaggct
     8821 tcccagtggc ctcaggagag tgacatgggg cagaaggtag cttctttgag tgcgcagtgc
     8881 cacaaccatg atgagagact tgcagagctg acagtcctgc ttcagaaact acagatacgg
     8941 gtagaccaag tggatgacgg cagggaaggg ctgtcactgt gggtcaagaa tgtggttgga
     9001 cagcacctgc aggagatggg caccatagaa ccacctgatg ctaagactga cttcatgact
     9061 ttccaccatg accatgaagt gcgtctctcc aacttggaag atgttcttag aaaactgaca
     9121 gaaaaatctg aggctatcca gaaggagctg gaagaaacca agctgaaagc aggcagcagg
     9181 gatgaagagc agcccctcct tgaccgtgtg cagcacctag aactggaact gaacctgttg
     9241 aagtcacagc tgtcagactg gcagcatctg aagaccagct gtgagcaggc tggggcccgc
     9301 atccaggaga ctgtgcagct catgttctct gaggatcagc agggcggttc cctcgagtgg
     9361 ctattagaga agctttcttc tcggttcgtg agcaaggatg agctgcaggt gctcttacat
     9421 gaccttgagc tgaaactgct gcagaatatc acacaccaca tcaccgtgac aggacaggcc
     9481 ccgacatccg aggctattgt gtctgccgtg aatcaggcag ggatttcagg aatcacagaa
     9541 gcgcaagcac atatcattgt gaacaatgct ctgaagctgt actcccaaga caagacgggg
     9601 atggtggact ttgctctgga gtctggaggt ggcagcatcc taagcactcg gtgctctgag
     9661 acctatgaga ccaagacggc actgctgagc ctgtttgggg tcccactgtg gtacttctca
     9721 cagtcacctc gagtggtgat ccagcccgac atctacccag ggaattgctg ggcgttcaaa
     9781 ggttcccagg ggtacctggt ggtgcggttg tccatgaaga tctacccaac cacattcacc
     9841 atggaacaca ttccaaagac actatcaccc actggtaaca tctccagtgc ccccaaagac
     9901 tttgcagtct atggactgga aacggagtat caagaagagg ggcagcctct gggacggttc
     9961 acctatgacc aggaaggaga ctcactccag atgttccaca cactggaaag acctgaccaa
    10021 gccttccaga tagtagagct ccgggtcctg tccaactggg gccaccctga gtacacttgc
    10081 ctctaccggt tccgagtcca cggagagccc atccagatga gcaaaggaga agaacttttc
    10141 actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct
    10201 gtccgtggag agggtgaagg tgatgctaca aacggaaaac tcacccttaa atttatttgc
    10261 actactggaa aactacctgt tccatggcca acacttgtca ctactctgac ctatggtgtt
    10321 caatgctttt cccgttatcc ggatcacatg aaacggcatg actttttcaa gagtgccatg
    10381 cccgaaggtt atgtacagga acgcactata tctttcaaag atgacgggac ctacaagacg
    10441 cgtgctgaag tcaagtttga aggtgatacc cttgttaatc gtatcgagtt aaaaggtatt
    10501 gattttaaag aagatggaaa cattctcgga cacaaactcg agtacaactt taactcacac
    10561 aatgtataca tcacggcaga caaacaaaag aatggaatca aagctaactt caaaattcgc
    10621 cacaacgttg aagatggttc cgttcaacta gcagaccatt atcaacaaaa tactccaatt
    10681 ggcgatggcc ctgtcctttt accagacaac cattacctgt cgacacaatc tgtcctttcg
    10741 aaagatccca acgaaaagcg tgaccacatg gtccttcttg agtttgtaac tgctgctggg
    10801 attacacatg gcatggatga gctctacaaa taa
//
