LOCUS       sequence-159109-       15002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for doxycycline-inducible expression of membrane 
            targeted-CSK in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15002)
  AUTHORS   Kim NG, Gumbiner BM
  TITLE     Adhesion to fibronectin regulates Hippo signaling via the 
            FAK-Src-PI3K pathway.
  JOURNAL   J Cell Biol. 2015 Aug 3;210(3):503-15. doi: 10.1083/jcb.201501025. 
            Epub 2015 Jul 27.
  PUBMED    26216901
REFERENCE   2  (bases 1 to 15002)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1083/jcb.201501025"; journalName: "J Cell Biol"; date: 
            "2015-08-3- 3"; volume: "210"; issue: "3"; pages: "503-15"
COMMENT     Sequence Label: Tet-myr-CSK-GFP
FEATURES             Location/Qualifiers
     source          1..15002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2063..2180
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(2245..2619)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     regulatory      2616..2625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2629..2647)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2639..2686)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    2722..2740
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2757..2775
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2793..2811
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2829..2847
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2864..2882
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2900..2918
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        2930..2968
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2965..2989
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3009..3033
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3033..3074
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     regulatory      4437..4446
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4443..4445,4446..4448,4449..5162)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4443..4445,4446..4448,4449..5159)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4488..4509)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4749..4768)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5096..5117
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5162..5186)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5194..5217
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5221..5244
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     ncRNA           5309..5430
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           5458..5472
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           5500..5626
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     promoter        complement(5721..5739)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        5752..6963
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6800..6819
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    complement(6976..7009)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     CDS             7016..7723
                     /codon_start=1
                     /product="reverse tetracycline transactivator 3"
                     /label=rtTA3
                     /note="activates expression from Tet-responsive promoters 
                     in the presence of doxycycline"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPG"
     protein_bind    complement(7724..7757)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     misc_feature    7780..8352
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7947..7964)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8174..8193
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8355..8957
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     8854..8874
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8973..9561
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9026..9046)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9444..9455)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9768..10001
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10102..10119)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10108..10332
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10378..10806
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10465..10484)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10675..10696
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10815..10835)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10820..11149
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11000..11135
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11062..11081
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    12357..12478
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12527..12543)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12527..12543)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12540..12562)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12551..12567
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(12575..12581,12582..12599,12600..12605))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12620..12641
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12758..12775)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12929..13517)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13009..13028)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13688..14479,14480..14548))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     14311..14330
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14549..14653)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    14701..14835
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
     2041 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     2101 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     2161 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag
     2221 taccgggccc gctctagtcc ggaatcagtc ctgctcctcg gccacgaagt gcacgcagtt
     2281 gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat
     2341 ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag
     2401 ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg
     2461 gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac
     2521 gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg
     2581 cgcggtgagc accggaacgg cactggtcaa cttggccatg gtggccctcc tatagtgagt
     2641 cgtattatac tatgccgata tactatgccg atgattaatt gtcaacacgt gctgcaggtc
     2701 cgaggttcta gacgagttta ctccctatca gtgatagaga acgatgtcga gtttactccc
     2761 tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc
     2821 gagtttactc cctatcagtg atagagaacg tatgtcgagt ttatccctat cagtgataga
     2881 gaacgtatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag gtaggcgtgt
     2941 acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagatcg caccggttgc
     3001 agatatcaac aagtttgtac aaaaaagttg gcatggggag cagcaagagc aagcccaagg
     3061 accccagcca gcgccggcgc aggatgtcag caatacaggc cgcctggcca tccggtacag
     3121 aatgtattgc caagtacaac ttccacggca ctgccgagca ggacctgccc ttctgcaaag
     3181 gagacgtgct caccattgtg gccgtcacca aggaccccaa ctggtacaaa gccaaaaaca
     3241 aggtgggccg tgagggcatc atcccagcca actacgtcca gaagcgggag ggcgtgaagg
     3301 cgggtaccaa actcagcctc atgccttggt tccacggcaa gatcacacgg gagcaggctg
     3361 agcggcttct gtacccgccg gagacaggcc tgttcctggt gcgggagagc accaactacc
     3421 ccggagacta cacgctgtgc gtgagctgcg acggcaaggt ggagcactac cgcatcatgt
     3481 accatgccag caagctcagc atcgacgagg aggtgtactt tgagaacctc atgcagctgg
     3541 tggagcacta cacctcagac gcagatggac tctgtacgcg cctcattaaa ccaaaggtca
     3601 tggagggcac agtggcggcc caggatgagt tctaccgcag cggctgggcc ctgaacatga
     3661 aggagctgaa gctgctgcag accatcggga agggggagtt cggagacgtg atgctgggcg
     3721 attaccgagg gaacaaagtc gccgtcaagt gcattaagaa cgacgccact gcccaggcct
     3781 tcctggctga agcctcagtc atgacgcaac tgcggcatag caacctggtg cagctcctgg
     3841 gcgtgatcgt ggaggagaag ggcgggctct acatcgtcac tgagtacatg gccaagggga
     3901 gccttgtgga ctacctgcgg tctaggggtc ggtcagtgct gggcggagac tgtctcctca
     3961 agttctcgct agatgtctgc gaggccatgg aatacctgga gggcaacaat ttcgtgcatc
     4021 gagacctggc tgcccgcaat gtgctggtgt ctgaggacaa cgtggccaag gtcagcgact
     4081 ttggtctcac caaggaggcg tccagcaccc aggacacggg caagctgcca gtcaagtgga
     4141 cagcccctga ggccctgaga gagaagaaat tctccactaa gtctgacgtg tggagtttcg
     4201 gaatccttct ctgggaaatc tactcctttg ggcgagtgcc ttatccaaga attcccctga
     4261 aggacgtcgt ccctcgggtg gagaagggct acaagatgga tgcccccgac ggctgcccgc
     4321 ccgcagtcta tgaagtcatg aagaactgct ggcacctgga cgccgccatg cggccctcct
     4381 tcctacagct ccgagagcag cttgagcaca tcaaaaccca cgagctgcac ctggcggccg
     4441 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     4501 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     4561 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     4621 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     4681 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     4741 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     4801 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     4861 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     4921 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     4981 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     5041 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     5101 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     5161 aacccaactt tcttgtacaa agtggttgat atcgactaca aggacgacga tgacaagatc
     5221 gactacaagg acgacgatga caagatcgac tacaaggacg acgatgacaa tccatcacac
     5281 tggcggccgc tctaaatcga tagtttgttt gaatgaggct tcagtacttt acagaatcgt
     5341 tgcctgcaca tcttggaaac acttgctggg attacttctt caggttaacc caacagaagg
     5401 ctcgagaagg tatattgctg ttgacagtga gcgatctcgc ttgggcgaga gtaagtagtg
     5461 aagccacaga tgtacttact ctcgcccaag cgagagtgcc tactgcctcg gaattcaagg
     5521 ggctacttta ggagcaatta tcttgtttac taaaactgaa taccttgcta tctctttgat
     5581 acatttttac aaagctgaat taaaatggta taaattaaat cacttttttc aattggaaga
     5641 ctaatgcggc cggccattac tccgtctcgt gtcttgttgc atatgtctgc tggtttgttt
     5701 gatgttgttt gcgggcgggc cctatagtga gtcgtattac ctaggacgcg tggcctccgc
     5761 gccgggtttt ggcgcctccc gcgggcgccc ccctcctcac ggcgagcgct gccacgtcag
     5821 acgaagggcg cagcgagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct
     5881 gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact
     5941 tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt
     6001 cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata
     6061 aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct
     6121 tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc
     6181 tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg
     6241 tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat
     6301 ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga
     6361 aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgaggcct tcgctaatgc
     6421 gggaaagctc ttattcgggt gagatgggct ggggcaccat ctggggaccc tgacgtgaag
     6481 tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt
     6541 gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca
     6601 cccgttctgt tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct
     6661 tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg
     6721 ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt
     6781 acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt
     6841 gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat
     6901 tttcagtgtt agactagtaa attgtccgct aaattctggc cgtttttggc ttttttgtta
     6961 gacgctagcg gatccataac ttcgtatagt atacattata cgaagttatg ccaccatgtc
     7021 taggctggac aagagcaaag tcataaacgg agctctggaa ttactcaatg gtgtcggtat
     7081 cgaaggcctg acgacaagga aactcgctca aaagctggga gttgagcagc ctaccctgta
     7141 ctggcacgtg aagaacaagc gggccctgct cgatgccctg ccaatcgaga tgctggacag
     7201 gcatcatacc cacttctgcc ccctggaagg cgagtcatgg caagactttc tgcggaacaa
     7261 cgccaagtca taccgctgtg ctctcctctc acatcgcgac ggggctaaag tgcatctcgg
     7321 cacccgccca acagagaaac agtacgaaac cctggaaaat cagctcgcgt tcctgtgtca
     7381 gcaaggcttc tccctggaga acgcactgta cgctctgtcc gccgtgggcc actttacact
     7441 gggctgcgta ttggaggaac aggagcatca agtagcaaaa gaggaaagag agacacctac
     7501 caccgattct atgcccccac ttctgagaca agcaattgag ctgttcgacc ggcagggagc
     7561 cgaacctgcc ttccttttcg gcctggaact aatcatatgt ggcctggaga aacagctaaa
     7621 gtgcgaaagc ggcgggccga ccgacgccct tgacgatttt gacttagaca tgctcccagc
     7681 cgatgccctt gacgattttg accttgacat gctccccggg taaataactt cgtatagtat
     7741 acattatacg aagttatgga tccgcggccg caaattccgc ccctctccct cccccccccc
     7801 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     7861 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     7921 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     7981 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     8041 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     8101 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     8161 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     8221 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     8281 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     8341 aacacgataa taccatggcc accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     8401 acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     8461 acaccgtcga cccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     8521 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     8581 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggctcgcgca
     8641 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     8701 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     8761 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgctg
     8821 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     8881 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     8941 gcaagcccgg tgcctgagtt cgcgtctgga acaatcaacc tctggattac aaaatttgtg
     9001 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     9061 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     9121 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     9181 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     9241 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     9301 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     9361 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     9421 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     9481 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     9541 ccctttgggc cgcctccccg cctggaatta attctgcagt cgagacctag aaaaacatgg
     9601 agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca
     9661 agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta
     9721 caaggcagct gtagatctta gccacttttt aaaagaaaag aggggactgg aagggctaat
     9781 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca
     9841 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     9901 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     9961 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
    10021 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggccttgaca
    10081 ttgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
    10141 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
    10201 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
    10261 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
    10321 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
    10381 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
    10441 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
    10501 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
    10561 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
    10621 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
    10681 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
    10741 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
    10801 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
    10861 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
    10921 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
    10981 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
    11041 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
    11101 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
    11161 agcttgtata tccattttcg gatctgatca gcacgtgatg aaaaagcctg aactcaccgc
    11221 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct
    11281 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct
    11341 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
    11401 atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattaattc agcgagagcc
    11461 tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg
    11521 aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc
    11581 ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat
    11641 ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg
    11701 acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg
    11761 actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg
    11821 acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat
    11881 acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc
    11941 gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc
    12001 tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag
    12061 cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta
    12121 cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg
    12181 atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac gtgctacgag
    12241 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
    12301 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
    12361 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
    12421 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
    12481 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
    12541 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
    12601 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
    12661 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
    12721 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
    12781 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
    12841 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
    12901 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
    12961 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
    13021 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
    13081 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
    13141 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
    13201 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
    13261 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
    13321 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
    13381 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
    13441 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
    13501 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
    13561 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
    13621 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
    13681 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
    13741 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
    13801 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
    13861 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
    13921 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
    13981 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
    14041 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
    14101 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
    14161 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
    14221 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
    14281 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
    14341 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
    14401 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
    14461 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
    14521 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
    14581 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
    14641 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
    14701 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
    14761 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
    14821 tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc
    14881 ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac ctgtgattcc
    14941 tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta
    15001 gt
//
