LOCUS       sequence-339473-       10989 bp    DNA     circular SYN 14-APR-2025
DEFINITION  lentiviral construct (3rd generation) for the expression of cell 
            cycle-independent OsTIR1 fused with mEmerald and miniAID-fused 
            mCherry2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10989)
  AUTHORS   Capece M, Tessari A, Mills J, Vinciguerra GLR, Louke D, Lin C, 
            McElwain BK, Miles WO, Coppola V, Davies AE, Palmieri D, Croce CM
  TITLE     A novel auxin-inducible degron system for rapid, cell cycle-specific
            targeted proteolysis.
  JOURNAL   Cell Death Differ. 2023 Aug 3. doi: 10.1038/s41418-023-01191-4.
  PUBMED    37537305
REFERENCE   2  (bases 1 to 10989)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Death 
            Differ. 2023 Aug 3. doi: 10.1038/s41418-023-01191-4."
COMMENT     Sequence Label: pLentiDP1.0
FEATURES             Location/Qualifiers
     source          1..10989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /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    1802..1918
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        1941..2244
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2245..2448
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2399..2419
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2486..4210
                     /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             join(4238..4240,4241..4243,4244..4954)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4283..4304)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4544..4563)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4891..4912
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4964..5020
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5021..5224
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     CDS             5225..5935
                     /codon_start=1
                     /product="derivative of the monomeric red fluorescent 
                     protein mCherry"
                     /label=mCherry2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFNWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQCRTMGWEASTERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVDIKLDILSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5370..5388)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    5936..6524
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5989..6009)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6407..6418)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6575..6595)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6580..6909
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6760..6895
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6957..7004
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7023..7421
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             7508..7741
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7813..7934
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7850..7869)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7904..7923
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7974..8109
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8129..8148)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8130..8148)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8158..8175)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8158..8174)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8167..8189)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8316..8771
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8403..8422)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8613..8634
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8797..8901
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8902..8970,8971..9762)
                     /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     complement(9120..9139)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9933..10521
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10422..10441
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10675..10692
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10809..10830
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10845..10850,10851..10868,10869..10875)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10883..10899
                     /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)."
     primer_bind     10888..10910
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10907..10923
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10907..10923
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10942..10962
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10944..10962
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     1741 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcggttaa
     1801 cttttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     1861 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaattc aaaattttat
     1921 cgataagctt gggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     1981 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     2041 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     2101 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     2161 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     2221 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2281 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2341 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2401 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
     2461 cgtcagatcc gctagcgctg ccaccatgac atactttcct gaagaggtcg tcgaacacat
     2521 ttttagcttc ctgcctgcac agagagatag aaacacagtg agcctggtct gcaaagtgtg
     2581 gtacgagatc gaacgcctga gccggagagg agtgttcgtc ggcaactgct atgctgtgag
     2641 agcaggcagg gtcgccgcta ggtttccaaa tgtgcgcgca ctgaccgtca aggggaaacc
     2701 ccacttcgcc gactttaacc tggtgccccc tgattgggga ggatacgccg gcccttggat
     2761 cgaggcagcc gctcgcggct gtcatggact ggaggaactg cgcatgaagc gaatggtggt
     2821 ctctgacgaa agtctggagc tgctggctcg gagcttccct aggtttcgcg cactggtgct
     2881 gatttcttgc gaaggcttca gcaccgatgg actggcagcc gtggcctccc actgtaagct
     2941 gctgcgggag ctggacctcc aggagaatga agtggaggat agaggcccca gatggctgtc
     3001 ttgcttccca gactcatgta ccagcctggt gtccctgaac tttgcctgca tcaaaggcga
     3061 agtgaatgct gggtccctgg agcggctggt ctcaagaagc cccaacctga ggtctctgcg
     3121 gctgaaccgg agcgtgagcg tggacactct ggctaagatt ctgctgagaa cccctaacct
     3181 ggaggatctg ggaaccggca atctgacaga cgatttccag acagaatcct actttaaact
     3241 gacttctgcc ctggagaagt gtaaaatgct gaggagtctg tcaggattct gggatgcttc
     3301 acccgtgtgc ctgagcttta tctaccctct gtgtgcacag ctgacaggcc tgaacctgag
     3361 ctatgcacca accctggacg ccagtgatct gacaaagatg atctcacgct gcgtgaaact
     3421 ccagcgactg tgggtgctgg actgtatttc cgataagggg ctccaggtgg tcgccagctc
     3481 ctgcaaggac ctccaggagc tgagagtgtt cccatctgat ttttacgtgg ccggatatag
     3541 tgctgtcact gaggaaggcc tggtggcagt ctcactggga tgcccaaagc tgaacagcct
     3601 gctgtatttc tgtcatcaga tgactaatgc tgcactggtg accgtcgcca agaactgccc
     3661 taatttcacc cgatttcggc tgtgtattct ggaaccaggc aaacccgacg tggtcacatc
     3721 ccagccactg gatgaagggt ttggagctat cgtgagagag tgcaagggac tccagaggct
     3781 gagcatttcc ggcctgctga cagacaaagt gttcatgtac atcggcaagt atgctaagca
     3841 gctggagatg ctgagcattg catttgccgg agactccgat aagggcatga tgcacgtgat
     3901 gaacgggtgt aagaatctgc gaaaactgga aatccgggac agccctttcg gggatgccgc
     3961 tctgctggga aactttgcca gatacgagac aatgaggagc ctgtggatgt ctagttgcaa
     4021 tgtgactctg aagggctgtc aggtcctggc tagtaaaatg cctatgctga acgtggaagt
     4081 cattaatgag cgggacgggt ctaacgaaat ggaggaaaat catggcgacc tgccaaaggt
     4141 ggagaaactg tatgtgtatc ggaccaccgc aggggcaaga gatgatgctc ccaactttgt
     4201 gaagattctg ggaggttctg gtggttctgg tggttccatg gtgagcaagg gcgaggagct
     4261 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     4321 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     4381 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tgacctacgg
     4441 cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     4501 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     4561 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     4621 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     4681 ccacaaggtc tatatcaccg ccgacaagca gaagaacggc atcaaggtga acttcaagac
     4741 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     4801 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct
     4861 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     4921 cgggatcact ctcggcatgg acgagctgta caagggcagc ggcgccacaa acttctctct
     4981 gctaaagcaa gcaggtgatg ttgaagaaaa ccccgggcct aaggagaaga gtgcttgtcc
     5041 taaagatcca gccaaacctc cggccaaggc acaagttgtg ggatggccac cggtgagatc
     5101 ataccggaag aacgtgatgg tttcctgcca aaaatcaagc ggtggcccgg aggcggcggc
     5161 gttcgtgaag gtatcaatgg acggagcacc gtacttgagg aaaatcgatt tgaggatgta
     5221 taaaatggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
     5281 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
     5341 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
     5401 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
     5461 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaattggga
     5521 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
     5581 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     5641 cgtaatgcag tgtcgtacca tgggctggga ggcctccact gagcggatgt accccgagga
     5701 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     5761 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
     5821 cgtcgacatc aagttggaca tcctttccca caacgaggac tacaccatcg tggaacagta
     5881 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtaaaatca
     5941 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     6001 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     6061 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     6121 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     6181 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     6241 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     6301 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     6361 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     6421 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     6481 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttctgcagat
     6541 atccggttag taatgagttt ggaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     6601 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     6661 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     6721 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     6781 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     6841 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     6901 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg
     6961 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     7021 ccatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa
     7081 tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg
     7141 gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg
     7201 tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg
     7261 gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc
     7321 tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga
     7381 ttcgtgaatt gctgccctct ggttatgtgt gggagggcta agcacaattc gagctcggta
     7441 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     7501 gggggactgg aagggctagc tcactcccaa cgaagacaag atctgctttt tgcttgtact
     7561 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     7621 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     7681 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     7741 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     7801 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     7861 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     7921 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     7981 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     8041 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     8101 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     8161 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     8221 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     8281 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     8341 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     8401 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     8461 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     8521 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     8581 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     8641 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     8701 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     8761 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     8821 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     8881 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     8941 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     9001 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     9061 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     9121 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     9181 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     9241 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     9301 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     9361 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     9421 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     9481 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     9541 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     9601 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     9661 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     9721 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     9781 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     9841 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     9901 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     9961 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
    10021 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
    10081 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
    10141 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
    10201 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
    10261 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
    10321 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
    10381 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
    10441 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
    10501 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
    10561 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
    10621 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
    10681 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
    10741 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
    10801 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
    10861 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
    10921 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg
    10981 caagcttaa
//
