LOCUS       sequence-339099-       11298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral construct (3rd generation) for the expression of cell 
            cycle-dependent OsTIR1 fused with mEmerald and Cdt1 (AKA ROLECCS G1)
            and miniAID-fused mCherry2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11298)
  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 11298)
  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"; journalName: "Cell Death 
            Differ. 2023 Aug 3. doi: 10.1038/s41418-023-01191-4."
COMMENT     Sequence Label: pLentiDP1.3
FEATURES             Location/Qualifiers
     source          1..11298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    47..172
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    665..898
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1083..1127
                     /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    1394..1510
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        1533..1836
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1837..2040
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1991..2011
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2078..3802
                     /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(3830..3832,3833..3835,3836..4546)
                     /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(3875..3896)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4136..4155)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4483..4504
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4583..4855
                     /codon_start=1
                     /product="degron consisting of residues 30-120 of human 
                     Cdt1 (Zielke and Edgar, 2015)"
                     /label=Cdt1 (30-120)
                     /translation="PSPARPALRAPASATSGSRKRARPPAAPGRDQARPPARRRLRLSV
                     DEVSSPSTPEAPDIPACPSPGQKIKKSTPAAGQPPHLTSAQDQDTI"
     CDS             4865..4921
                     /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             4922..5125
                     /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             5126..5836
                     /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(5271..5289)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    5837..6425
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5890..5910)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6308..6319)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6476..6496)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6481..6810
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6661..6796
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6858..6905
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6924..7322
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             7409..7642
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7714..7835
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7751..7770)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7805..7824
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7875..8010
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8030..8049)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8031..8049)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8059..8076)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8059..8075)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8068..8090)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8217..8672
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8304..8323)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8514..8535
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8698..8802
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8803..8871,8872..9663)
                     /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(9021..9040)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9834..10422
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10323..10342
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10576..10593
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10710..10731
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10746..10751,10752..10769,10770..10776)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10784..10800
                     /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     10789..10811
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10808..10824
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10808..10824
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10843..10863
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10845..10863
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        10891..11117
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             11118..11298
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
       61 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
      121 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
      181 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
      241 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
      301 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
      361 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
      421 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
      481 aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag
      541 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
      601 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
      661 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
      721 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
      781 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
      841 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
      901 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
      961 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     1021 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     1081 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     1141 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     1201 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     1261 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     1321 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac
     1381 ggtatcggtt aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
     1441 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat
     1501 tcaaaatttt atcgataagc ttgggagttc cgcgttacat aacttacggt aaatggcccg
     1561 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     1621 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     1681 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     1741 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     1801 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     1861 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     1921 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     1981 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     2041 ggtttagtga accgtcagat ccgctagcgc tgccacaatg acatactttc ctgaagaggt
     2101 cgtcgaacac atttttagct tcctgcctgc acagagagat agaaacacag tgagcctggt
     2161 ctgcaaagtg tggtacgaga tcgaacgcct gagccggaga ggagtgttcg tcggcaactg
     2221 ctatgctgtg agagcaggca gggtcgccgc taggtttcca aatgtgcgcg cactgaccgt
     2281 caaggggaaa ccccacttcg ccgactttaa cctggtgccc cctgattggg gaggatacgc
     2341 cggcccttgg atcgaggcag ccgctcgcgg ctgtcatgga ctggaggaac tgcgcatgaa
     2401 gcgaatggtg gtctctgacg aaagtctgga gctgctggct cggagcttcc ctaggtttcg
     2461 cgcactggtg ctgatttctt gcgaaggctt cagcaccgat ggactggcag ccgtggcctc
     2521 ccactgtaag ctgctgcggg agctggacct ccaggagaat gaagtggagg atagaggccc
     2581 cagatggctg tcttgcttcc cagactcatg taccagcctg gtgtccctga actttgcctg
     2641 catcaaaggc gaagtgaatg ctgggtccct ggagcggctg gtctcaagaa gccccaacct
     2701 gaggtctctg cggctgaacc ggagcgtgag cgtggacact ctggctaaga ttctgctgag
     2761 aacccctaac ctggaggatc tgggaaccgg caatctgaca gacgatttcc agacagaatc
     2821 ctactttaaa ctgacttctg ccctggagaa gtgtaaaatg ctgaggagtc tgtcaggatt
     2881 ctgggatgct tcacccgtgt gcctgagctt tatctaccct ctgtgtgcac agctgacagg
     2941 cctgaacctg agctatgcac caaccctgga cgccagtgat ctgacaaaga tgatctcacg
     3001 ctgcgtgaaa ctccagcgac tgtgggtgct ggactgtatt tccgataagg ggctccaggt
     3061 ggtcgccagc tcctgcaagg acctccagga gctgagagtg ttcccatctg atttttacgt
     3121 ggccggatat agtgctgtca ctgaggaagg cctggtggca gtctcactgg gatgcccaaa
     3181 gctgaacagc ctgctgtatt tctgtcatca gatgactaat gctgcactgg tgaccgtcgc
     3241 caagaactgc cctaatttca cccgatttcg gctgtgtatt ctggaaccag gcaaacccga
     3301 cgtggtcaca tcccagccac tggatgaagg gtttggagct atcgtgagag agtgcaaggg
     3361 actccagagg ctgagcattt ccggcctgct gacagacaaa gtgttcatgt acatcggcaa
     3421 gtatgctaag cagctggaga tgctgagcat tgcatttgcc ggagactccg ataagggcat
     3481 gatgcacgtg atgaacgggt gtaagaatct gcgaaaactg gaaatccggg acagcccttt
     3541 cggggatgcc gctctgctgg gaaactttgc cagatacgag acaatgagga gcctgtggat
     3601 gtctagttgc aatgtgactc tgaagggctg tcaggtcctg gctagtaaaa tgcctatgct
     3661 gaacgtggaa gtcattaatg agcgggacgg gtctaacgaa atggaggaaa atcatggcga
     3721 cctgccaaag gtggagaaac tgtatgtgta tcggaccacc gcaggggcaa gagatgatgc
     3781 tcccaacttt gtgaagattc tgggaggttc tggtggttct ggtggttcca tggtgagcaa
     3841 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     3901 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     3961 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     4021 cttgacctac ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     4081 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     4141 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     4201 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     4261 caactacaac agccacaagg tctatatcac cgccgacaag cagaagaacg gcatcaaggt
     4321 gaacttcaag acccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     4381 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     4441 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     4501 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagccta ggggatatcc
     4561 atcacactgg cggccgctcg agcccagccc cgccaggccc gcactccgcg ccccggcctc
     4621 cgctaccagt ggcagccgca agcgcgcccg cccgcccgcc gcccccggac gcgaccaggc
     4681 caggccaccg gcccgcagga gactgcggct gtcggtggac gaggtttcca gccccagtac
     4741 ccccgaggcc ccagacatcc cagcctgccc ttctccgggc cagaagataa agaaatccac
     4801 cccggcagca ggtcagccgc cccacctgac atccgcgcag gaccaggaca ccatcggcag
     4861 cggcgccaca aacttctctc tgctaaagca agcaggtgat gttgaagaaa accccgggcc
     4921 taaggagaag agtgcttgtc ctaaagatcc agccaaacct ccggccaagg cacaagttgt
     4981 gggatggcca ccggtgagat cataccggaa gaacgtgatg gtttcctgcc aaaaatcaag
     5041 cggtggcccg gaggcggcgg cgttcgtgaa ggtatcaatg gacggagcac cgtacttgag
     5101 gaaaatcgat ttgaggatgt ataaaatggt gagcaagggc gaggaggata acatggccat
     5161 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     5221 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     5281 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     5341 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     5401 ccccgagggc ttcaattggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     5461 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
     5521 caacttcccc tccgacggcc ccgtaatgca gtgtcgtacc atgggctggg aggcctccac
     5581 tgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
     5641 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
     5701 gcagctgccc ggcgcctaca acgtcgacat caagttggac atcctttccc acaacgagga
     5761 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
     5821 cgagctgtac aagtaaaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     5881 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     5941 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     6001 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     6061 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     6121 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     6181 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     6241 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     6301 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     6361 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     6421 cccgcctgga attctgcaga tatccggtta gtaatgagtt tggaattaat tctgtggaat
     6481 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     6541 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     6601 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     6661 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     6721 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     6781 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     6841 ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat
     6901 acgacaaggt gaggaactaa accatggcca agcctttgtc tcaagaagaa tccaccctca
     6961 ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca
     7021 gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg
     7081 ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc
     7141 tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt
     7201 gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg
     7261 gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct
     7321 aagcacaatt cgagctcggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     7381 agccactttt taaaagaaaa ggggggactg gaagggctag ctcactccca acgaagacaa
     7441 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     7501 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     7561 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     7621 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
     7681 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     7741 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     7801 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     7861 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     7921 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     7981 agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga
     8041 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     8101 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     8161 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
     8221 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     8281 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     8341 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     8401 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     8461 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     8521 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     8581 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     8641 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     8701 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     8761 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     8821 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     8881 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     8941 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     9001 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     9061 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     9121 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     9181 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     9241 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     9301 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     9361 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     9421 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     9481 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     9541 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     9601 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     9661 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     9721 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     9781 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     9841 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     9901 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     9961 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
    10021 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
    10081 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
    10141 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
    10201 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
    10261 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
    10321 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
    10381 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
    10441 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
    10501 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
    10561 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
    10621 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
    10681 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
    10741 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
    10801 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
    10861 agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt
    10921 gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat
    10981 gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc
    11041 tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc
    11101 tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg
    11161 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
    11221 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
    11281 tgtggaaaat ctctagca
//
