LOCUS       sequence-340211-       14360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for the Tet-inducible expression of HA-tagged TIR1
            and a constitutive puromycin resistance marker in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14360)
  AUTHORS   Simoneschi D, Rona G, Zhou N, Jeong YT, Jiang S, Milletti G, Arbini 
            AA, O'Sullivan A, Wang AA, Nithikasem S, Keegan S, Siu Y, 
            Cianfanelli V, Maiani E, Nazio F, Cecconi F, Boccalatte F, Fenyo D, 
            Jones DR, Busino L, Pagano M
  TITLE     CRL4(AMBRA1) is a master regulator of D-type cyclins.
  JOURNAL   Nature. 2021 Apr;592(7856):789-793. doi: 10.1038/s41586-021-03445-y.
            Epub 2021 Apr 14.
  PUBMED    33854235
REFERENCE   2  (bases 1 to 14360)
  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: 
            "10.1038/s41586-021-03445-y"; journalName: "Nature"; date: 
            "2021-04"; volume: "592"; issue: "7856"; pages: "789-793"
COMMENT     Sequence Label: pTRIPZ-TIR1-HA
FEATURES             Location/Qualifiers
     source          1..14360
                     /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"
     CDS             3004..4728
                     /codon_start=1
                     /product="rice transport inhibitor response 1 protein"
                     /label=OsTIR1
                     /note="F-box protein that mediates auxin-dependent 
                     interaction of a ubiquitin ligase with an auxin-inducible 
                     degron (Tan et al., 2007)"
                     /translation="MTYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFV
                     GNCYAVRAGRVAARFPNVRALTVKGKPHFADFNLVPPDWGGYAGPWIEAAARGCHGLEE
                     LRMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEV
                     EDRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTL
                     AKILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYP
                     LCAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELR
                     VFPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRL
                     CILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSI
                     AFAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKG
                     CQVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKIL"
     CDS             4729..4755
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             4729..4755
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     ncRNA           4816..4830
                     /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           4858..4984
                     /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(5079..5097)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        5110..6321
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6158..6177
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    complement(6334..6367)
                     /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             6374..7081
                     /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(7082..7115)
                     /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    7138..7710
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7305..7322)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7532..7551
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7713..8315
                     /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     8212..8232
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8331..8919
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8384..8404)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8802..8813)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9126..9359
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9460..9477)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9466..9690
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9736..10164
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9823..9842)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10033..10054
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10173..10193)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10178..10507
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10358..10493
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10420..10439
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    11715..11836
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11885..11901)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11885..11901)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11898..11920)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11909..11925
                     /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(11933..11939,11940..11957,11958..11963))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11978..11999
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12116..12133)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12287..12875)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12367..12386)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13046..13837,13838..13906))
                     /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     13669..13688
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13907..14011)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    14059..14193
                     /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 caccggtgcc
     3001 accatgacat actttcctga agaggtcgtc gaacacattt ttagcttcct gcctgcacag
     3061 agagatagaa acacagtgag cctggtctgc aaagtgtggt acgagatcga acgcctgagc
     3121 cggagaggag tgttcgtcgg caactgctat gctgtgagag caggcagggt cgccgctagg
     3181 tttccaaatg tgcgcgcact gaccgtcaag gggaaacccc acttcgccga ctttaacctg
     3241 gtgccccctg attggggagg atacgccggc ccttggatcg aggcagccgc tcgcggctgt
     3301 catggactgg aggaactgcg catgaagcga atggtggtct ctgacgaaag tctggagctg
     3361 ctggctcgga gcttccctag gtttcgcgca ctggtgctga tttcttgcga aggcttcagc
     3421 accgatggac tggcagccgt ggcctcccac tgtaagctgc tgcgggagct ggacctccag
     3481 gagaatgaag tggaggatag aggccccaga tggctgtctt gcttcccaga ctcatgtacc
     3541 agcctggtgt ccctgaactt tgcctgcatc aaaggcgaag tgaatgctgg gtccctggag
     3601 cggctggtct caagaagccc caacctgagg tctctgcggc tgaaccggag cgtgagcgtg
     3661 gacactctgg ctaagattct gctgagaacc cctaacctgg aggatctggg aaccggcaat
     3721 ctgacagacg atttccagac agaatcctac tttaaactga cttctgccct ggagaagtgt
     3781 aaaatgctga ggagtctgtc aggattctgg gatgcttcac ccgtgtgcct gagctttatc
     3841 taccctctgt gtgcacagct gacaggcctg aacctgagct atgcaccaac cctggacgcc
     3901 agtgatctga caaagatgat ctcacgctgc gtgaaactcc agcgactgtg ggtgctggac
     3961 tgtatttccg ataaggggct ccaggtggtc gccagctcct gcaaggacct ccaggagctg
     4021 agagtgttcc catctgattt ttacgtggcc ggatatagtg ctgtcactga ggaaggcctg
     4081 gtggcagtct cactgggatg cccaaagctg aacagcctgc tgtatttctg tcatcagatg
     4141 actaatgctg cactggtgac cgtcgccaag aactgcccta atttcacccg atttcggctg
     4201 tgtattctgg aaccaggcaa acccgacgtg gtcacatccc agccactgga tgaagggttt
     4261 ggagctatcg tgagagagtg caagggactc cagaggctga gcatttccgg cctgctgaca
     4321 gacaaagtgt tcatgtacat cggcaagtat gctaagcagc tggagatgct gagcattgca
     4381 tttgccggag actccgataa gggcatgatg cacgtgatga acgggtgtaa gaatctgcga
     4441 aaactggaaa tccgggacag ccctttcggg gatgccgctc tgctgggaaa ctttgccaga
     4501 tacgagacaa tgaggagcct gtggatgtct agttgcaatg tgactctgaa gggctgtcag
     4561 gtcctggcta gtaaaatgcc tatgctgaac gtggaagtca ttaatgagcg ggacgggtct
     4621 aacgaaatgg aggaaaatca tggcgacctg ccaaaggtgg agaaactgta tgtgtatcgg
     4681 accaccgcag gggcaagaga tgatgctccc aactttgtga agattctgta cccctacgac
     4741 gtgcccgact acgcctgact cgagaaggta tattgctgtt gacagtgagc gatctcgctt
     4801 gggcgagagt aagtagtgaa gccacagatg tacttactct cgcccaagcg agagtgccta
     4861 ctgcctcgga attcaagggg ctactttagg agcaattatc ttgtttacta aaactgaata
     4921 ccttgctatc tctttgatac atttttacaa agctgaatta aaatggtata aattaaatca
     4981 cttttttcaa ttggaagact aatgcggccg gccattactc cgtctcgtgt cttgttgcat
     5041 atgtctgctg gtttgtttga tgttgtttgc gggcgggccc tatagtgagt cgtattacct
     5101 aggacgcgtg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcctcacgg
     5161 cgagcgctgc cacgtcagac gaagggcgca gcgagcgtcc tgatccttcc gcccggacgc
     5221 tcaggacagc ggcccgctgc tcataagact cggccttaga accccagtat cagcagaagg
     5281 acattttagg acgggacttg ggtgactcta gggcactggt tttctttcca gagagcggaa
     5341 caggcgagga aaagtagtcc cttctcggcg attctgcgga gggatctccg tggggcggtg
     5401 aacgccgatg attatataag gacgcgccgg gtgtggcaca gctagttccg tcgcagccgg
     5461 gatttgggtc gcggttcttg tttgtggatc gctgtgatcg tcacttggtg agtagcgggc
     5521 tgctgggctg gccggggctt tcgtggccgc cgggccgctc ggtgggacgg aagcgtgtgg
     5581 agagaccgcc aagggctgta gtctgggtcc gcgagcaagg ttgccctgaa ctgggggttg
     5641 gggggagcgc agcaaaatgg cggctgttcc cgagtcttga atggaagacg cttgtgaggc
     5701 gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg gcggcaagaa cccaaggtct
     5761 tgaggccttc gctaatgcgg gaaagctctt attcgggtga gatgggctgg ggcaccatct
     5821 ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg
     5881 ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag cgcgcgccct
     5941 cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg ccacctgccg
     6001 gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct agggtaggct
     6061 ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag gcgtcagttt
     6121 ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt ttgaactatg
     6181 cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga aatgtaatca
     6241 tttgggtcaa tatgtaattt tcagtgttag actagtaaat tgtccgctaa attctggccg
     6301 tttttggctt ttttgttaga cgctagcgga tccataactt cgtatagtat acattatacg
     6361 aagttatgcc accatgtcta ggctggacaa gagcaaagtc ataaacggag ctctggaatt
     6421 actcaatggt gtcggtatcg aaggcctgac gacaaggaaa ctcgctcaaa agctgggagt
     6481 tgagcagcct accctgtact ggcacgtgaa gaacaagcgg gccctgctcg atgccctgcc
     6541 aatcgagatg ctggacaggc atcataccca cttctgcccc ctggaaggcg agtcatggca
     6601 agactttctg cggaacaacg ccaagtcata ccgctgtgct ctcctctcac atcgcgacgg
     6661 ggctaaagtg catctcggca cccgcccaac agagaaacag tacgaaaccc tggaaaatca
     6721 gctcgcgttc ctgtgtcagc aaggcttctc cctggagaac gcactgtacg ctctgtccgc
     6781 cgtgggccac tttacactgg gctgcgtatt ggaggaacag gagcatcaag tagcaaaaga
     6841 ggaaagagag acacctacca ccgattctat gcccccactt ctgagacaag caattgagct
     6901 gttcgaccgg cagggagccg aacctgcctt ccttttcggc ctggaactaa tcatatgtgg
     6961 cctggagaaa cagctaaagt gcgaaagcgg cgggccgacc gacgcccttg acgattttga
     7021 cttagacatg ctcccagccg atgcccttga cgattttgac cttgacatgc tccccgggta
     7081 aataacttcg tatagtatac attatacgaa gttatggatc cgcggccgca aattccgccc
     7141 ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     7201 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     7261 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     7321 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     7381 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     7441 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     7501 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     7561 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
     7621 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     7681 cgtggttttc ctttgaaaaa cacgataata ccatggccac cgagtacaag cccacggtgc
     7741 gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg
     7801 actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc
     7861 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     7921 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     7981 ccgagatcgg ctcgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     8041 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     8101 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     8161 aggcggccga gcgcgctggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     8221 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     8281 gcacctggtg catgacccgc aagcccggtg cctgagttcg cgtctggaac aatcaacctc
     8341 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     8401 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     8461 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     8521 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     8581 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     8641 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     8701 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
     8761 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     8821 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     8881 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattaat tctgcagtcg
     8941 agacctagaa aaacatggag caatcacaag tagcaataca gcagctacca atgctgattg
     9001 tgcctggcta gaagcacaag aggaggagga ggtgggtttt ccagtcacac ctcaggtacc
     9061 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaagag
     9121 gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg
     9181 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     9241 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     9301 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     9361 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     9421 gagtgagagg ccttgacatt gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     9481 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     9541 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     9601 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     9661 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     9721 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     9781 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     9841 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     9901 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     9961 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
    10021 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
    10081 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
    10141 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
    10201 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
    10261 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
    10321 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
    10381 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
    10441 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
    10501 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgatgaa
    10561 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
    10621 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
    10681 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
    10741 tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga
    10801 attaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc
    10861 aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg
    10921 cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa
    10981 tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc
    11041 actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc
    11101 tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct
    11161 ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga
    11221 tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt
    11281 gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc
    11341 ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg
    11401 gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag
    11461 ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct
    11521 gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg
    11581 aatagcacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
    11641 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
    11701 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
    11761 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
    11821 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
    11881 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
    11941 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
    12001 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
    12061 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
    12121 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
    12181 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
    12241 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
    12301 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
    12361 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
    12421 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
    12481 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
    12541 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
    12601 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
    12661 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
    12721 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
    12781 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
    12841 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
    12901 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
    12961 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
    13021 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
    13081 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
    13141 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
    13201 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
    13261 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
    13321 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
    13381 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
    13441 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
    13501 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
    13561 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
    13621 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
    13681 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
    13741 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
    13801 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
    13861 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
    13921 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
    13981 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
    14041 tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa ataaagcaat
    14101 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
    14161 aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat
    14221 catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta
    14281 ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata
    14341 tgtactacaa acttagtagt
//
