LOCUS       Exported               15015 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Lentiviral vector for Tet-inducible Ajuba expression.
ACCESSION   .
VERSION     .
KEYWORDS    pTRIPZ-EGFP:Ajuba
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15015)
  AUTHORS   Ibar C, Kirichenko E, Keepers B, Enners E, Fleisch K, Irvine KD
  TITLE     Tension-dependent regulation of mammalian Hippo signaling through 
            LIMD1.
  JOURNAL   J Cell Sci. 2018 Feb 13. pii: jcs.214700. doi: 10.1242/jcs.214700.
  PUBMED    29440237
REFERENCE   2  (bases 1 to 15015)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..15015
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     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"
     CDS             2993..3709
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2993..3709
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3038..3059)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3299..3318)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3646..3667
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     ncRNA           5408..5501
                     /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           5527..5639
                     /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"
     primer_bind     complement(5733..5752)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5734..5752)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        5765..6976
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6813..6832
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    6989..7022
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GTATACAT). loxP511 sites are compatible with each
                     other, but incompatible with wild-type loxP sites."
     CDS             7029..7736
                     /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    7737..7770
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GTATACAT). loxP511 sites are compatible with each
                     other, but incompatible with wild-type loxP sites."
     misc_feature    7793..8365
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7960..7977)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8187..8206
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8368..8970
                     /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     8867..8887
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8986..9574
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9039..9059)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9457..9468)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9781..10014
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10115..10132)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10121..10345
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10391..10819
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10478..10497)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10688..10709
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10828..10848)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10833..11162
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11013..11148
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11075..11094
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    12370..12491
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12540..12556)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12540..12556)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12553..12575)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12564..12580
                     /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(12588..12618)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12633..12654
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12771..12788)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12942..13530)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13022..13041)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(13701..14561)
                     /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     14324..14343
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14562..14666)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    14714..14848
                     /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 caatggtgag
     3001 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     3061 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     3121 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     3181 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     3241 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     3301 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     3361 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     3421 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     3481 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     3541 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     3601 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     3661 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt actcagatct
     3721 cgagatggag cggttaggag agaaagccag tcgcctgctg gagaagttcg gccgcagaaa
     3781 gggtgaatct agccggtctg ggtctgacgg gacccccggg ccgggcaagg ggcgcctaag
     3841 tgggttgggg ggacctagga agtcagggcc ccgaggagct actgggggac ctggggatga
     3901 gccgttggag ccggcccggg agcaaggttc cctggacgct gagcgaaatc agcgcggctc
     3961 ctttgaggcg ccgcgctacg aaggctcttt tcccgcgggg ccgccgccca cccgggcctt
     4021 gcctctacct cagtcgttgc cccccgattt tcggctggag cccacggccc cggccctcag
     4081 cccccgctct agcttcgcca gtagctcggc cagcgacgcg agcaagccgt ccagcccccg
     4141 gggcagcctg ctgctggacg gggcgggggc tggcggagct ggaggtagcc ggccctgcag
     4201 caatcgcacc agcggcatca gcatgggcta cgaccagcgc cacgggagcc ccttgccagc
     4261 ggggccgtgc ctgtttggcc cacccctggc cggagcaccg gcaggctatt ctcccggagg
     4321 ggtcccgtcc gcctacccgg agctccacgc cgccctggac cgattgtacg ctcagcggcc
     4381 cgcggggttc ggctgccagg aaagccgcca ctcgtatccc ccggccctgg gcagccctgg
     4441 agctctagcc ggggccggag tgggagcggc ggggcccttg gagagacggg gggcgcaacc
     4501 cggacgacac tctgtgaccg gctacgggga ctgcgccgtg ggcgcccggt accaggacga
     4561 gctaacagct ttgcttcgcc tgacggtggg caccggtggg cgagaagccg gagcccgcgg
     4621 agaaccctcg gggattgagc cgtcgggtct ggaggagcca ccaggtcctt tcgttccgga
     4681 ggccgcccgg gcccggatgc gggagccaga ggccagggag gactacttcg gcacctgtat
     4741 caagtgcaac aaaggcatct atgggcagag caatgcctgc caggccctgg acagcctcta
     4801 ccacacccag tgctttgttt gctgctcttg tgggcgaact ttgcgttgca aggctttcta
     4861 cagtgtcaat ggctctgtgt actgtgagga agattatctg ttttcagggt ttcaggaggc
     4921 agctgagaaa tgctgtgtct gtggtcactt gattttggag aagatcctac aagcaatggg
     4981 gaagtcctat catccaggct gtttccgatg cattgtttgc aacaagtgcc tggatggcat
     5041 ccccttcaca gtggacttct ccaaccaagt atactgtgtc accgactacc acaaaaatta
     5101 tgctcctaag tgtgcagcct gtggccaacc catcctcccc tctgagggct gtgaggacat
     5161 cgtgagggtg atatccatgg accgggatta tcactttgag tgctaccact gtgaggactg
     5221 ccggatgcag ctgagtgatg aggaaggctg ctgctgtttc cctctggatg ggcacttgct
     5281 ctgccatggt tgccacatgc agcggctcaa tgcccgacaa ccccctgcca actatatctg
     5341 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaat
     5401 agtttgtttg aatgaggctt cagtacttta cagaatcgtt gcctgcacat cttggaaaca
     5461 cttgctggga ttacttcttc aggttaaccc aacagaaggc tcgaggtaac cggatcctga
     5521 tcagaattca aggggctact ttaggagcaa ttatcttgtt tactaaaact gaataccttg
     5581 ctatctcttt gatacatttt tacaaagctg aattaaaatg gtataaatta aatcactttt
     5641 ttcaattgga agactaatgc ggccggccat tactccgtct cgtgtcttgt tgcatatgtc
     5701 tgctggtttg tttgatgttg tttgcgggcg ggccctatag tgagtcgtat tacctaggac
     5761 gcgtggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcct cacggcgagc
     5821 gctgccacgt cagacgaagg gcgcagcgag cgtcctgatc cttccgcccg gacgctcagg
     5881 acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt
     5941 ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc
     6001 gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc
     6061 cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt
     6121 gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg
     6181 ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg tgtggagaga
     6241 ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg ggttgggggg
     6301 agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt gaggcgggct
     6361 gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa ggtcttgagg
     6421 ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctggggcac catctgggga
     6481 ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg
     6541 cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg
     6601 tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg
     6661 tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct
     6721 gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg
     6781 gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc
     6841 ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg
     6901 gtcaatatgt aattttcagt gttagactag taaattgtcc gctaaattct ggccgttttt
     6961 ggcttttttg ttagacgcta gcggatccat aacttcgtat agtatacatt atacgaagtt
     7021 atgccaccat gtctaggctg gacaagagca aagtcataaa cggagctctg gaattactca
     7081 atggtgtcgg tatcgaaggc ctgacgacaa ggaaactcgc tcaaaagctg ggagttgagc
     7141 agcctaccct gtactggcac gtgaagaaca agcgggccct gctcgatgcc ctgccaatcg
     7201 agatgctgga caggcatcat acccacttct gccccctgga aggcgagtca tggcaagact
     7261 ttctgcggaa caacgccaag tcataccgct gtgctctcct ctcacatcgc gacggggcta
     7321 aagtgcatct cggcacccgc ccaacagaga aacagtacga aaccctggaa aatcagctcg
     7381 cgttcctgtg tcagcaaggc ttctccctgg agaacgcact gtacgctctg tccgccgtgg
     7441 gccactttac actgggctgc gtattggagg aacaggagca tcaagtagca aaagaggaaa
     7501 gagagacacc taccaccgat tctatgcccc cacttctgag acaagcaatt gagctgttcg
     7561 accggcaggg agccgaacct gccttccttt tcggcctgga actaatcata tgtggcctgg
     7621 agaaacagct aaagtgcgaa agcggcgggc cgaccgacgc ccttgacgat tttgacttag
     7681 acatgctccc agccgatgcc cttgacgatt ttgaccttga catgctcccc gggtaaataa
     7741 cttcgtatag tatacattat acgaagttat ggatccgcgg ccgcaaattc cgcccctctc
     7801 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     7861 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     7921 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     7981 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     8041 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     8101 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     8161 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     8221 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
     8281 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     8341 ttttcctttg aaaaacacga taataccatg gccaccgagt acaagcccac ggtgcgcctc
     8401 gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac
     8461 cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac cgagctgcaa
     8521 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     8581 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     8641 atcggctcgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     8701 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     8761 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     8821 gccgagcgcg ctggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     8881 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     8941 tggtgcatga cccgcaagcc cggtgcctga gttcgcgtct ggaacaatca acctctggat
     9001 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     9061 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     9121 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     9181 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     9241 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     9301 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     9361 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     9421 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     9481 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     9541 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc
     9601 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct
     9661 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa
     9721 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aagaggggac
     9781 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc
     9841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     9901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     9961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
    10021 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
    10081 agaggccttg acattgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
    10141 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
    10201 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
    10261 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
    10321 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
    10381 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
    10441 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
    10501 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
    10561 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
    10621 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
    10681 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
    10741 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
    10801 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
    10861 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
    10921 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
    10981 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
    11041 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
    11101 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
    11161 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg atgaaaaagc
    11221 ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg
    11281 acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc
    11341 gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt
    11401 atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattaa
    11461 ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac
    11521 ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc
    11581 gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt
    11641 caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg
    11701 caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg
    11761 ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac
    11821 aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc
    11881 ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg
    11941 gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc
    12001 cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat
    12061 ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg
    12121 actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta
    12181 gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag
    12241 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
    12301 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
    12361 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
    12421 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
    12481 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
    12541 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
    12601 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
    12661 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
    12721 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
    12781 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
    12841 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
    12901 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
    12961 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
    13021 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
    13081 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
    13141 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
    13201 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
    13261 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
    13321 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
    13381 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
    13441 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
    13501 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
    13561 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
    13621 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
    13681 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
    13741 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
    13801 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
    13861 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
    13921 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
    13981 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
    14041 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
    14101 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
    14161 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
    14221 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
    14281 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
    14341 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
    14401 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
    14461 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
    14521 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
    14581 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
    14641 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
    14701 ggatcgggag atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
    14761 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
    14821 catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc
    14881 caaacttccc accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta
    14941 aacctgtgat tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac
    15001 tacaaactta gtagt
//
