LOCUS       sequence-373726-       11925 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Doxycycline-inducible lentivirus vector to express 
            EB3N-VVDfast-mCl3-iLID, a microtubule anchor used for optogenetic 
            recruitment of SspB-mCh-p60..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11925)
  AUTHORS   Meiring JCM, Grigoriev I, Nijenhuis W, Kapitein LC, Akhmanova A
  TITLE     Opto-katanin, an optogenetic tool for localized, microtubule 
            disassembly.
  JOURNAL   Curr Biol. 2022 Nov 7;32(21):4660-4674.e6. doi: 
            10.1016/j.cub.2022.09.010. Epub 2022 Sep 28.
  PUBMED    36174574
REFERENCE   2  (bases 1 to 11925)
  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"; doi: 
            "10.1016/j.cub.2022.09.010"; journalName: "Curr Biol"; date: 
            "2022-11-7- 7"; volume: "32"; issue: "21"; pages: "4660-4674.e6"
COMMENT     Sequence Label: pSIN-TRE-EB3N-VVDfast-mCl3-iLID
FEATURES             Location/Qualifiers
     source          1..11925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /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    1804..1921
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    join(1954..1972,1973..1995,1996..2014,2015..2037,2038..2056,
                     2057..2079,2080..2098,2099..2121,2122..2140,2141..2163,
                     2164..2182,2183..2205,2206..2224)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2257..2295
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2292..2316
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(3508..3510,3511..3513,3514..4224)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(3553..3574)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     4161..4182
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4324..4761
                     /codon_start=1
                     /product="improved light-inducible dimer when used with 
                     SspB nano or SspB micro (Guntas et al., 2015)"
                     /label=iLID
                     /translation="LATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNVFHLQPMRDYKGDVQ
                     YFIGVQLDGTERLHGAAEREAVCLIKKTAFQIAEAANDENYF"
     promoter        4826..5336
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     5255..5273
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             join(5374..5991,5992..5997,5998..6033,6034..6036,6037..6072,
                     6073..6075,6076..6111,6112..6120)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    join(6129..6703,6704..6706,6707..6715)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6296..6313)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6523..6542
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6735..7760
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             6735..7760
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     misc_feature    7792..8380
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7845..7865)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8263..8274)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8467..8700
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8772..8893
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8809..8828)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8863..8882
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8912..9047
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9067..9086)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9068..9086)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9096..9113)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9096..9112)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9105..9127)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9254..9709
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9341..9360)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9551..9572
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9735..9839
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9840..9908,9909..10700)
                     /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(10058..10077)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10871..11459
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11360..11379
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11613..11630
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11747..11768
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11783..11788,11789..11806,11807..11813)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11821..11837
                     /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     11826..11848
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11845..11861
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11845..11861
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11880..11900
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11882..11900
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgatcac gagactagcc tcgagtttac cactccctat cagtgataga gaaaagtgaa
     1981 agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
     2041 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
     2101 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
     2161 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata
     2221 gagaaaagtg aaagtcgagc tcggtacccg ggtcgaggta ggcgtgtacg gtgggaggcc
     2281 tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt
     2341 tttgacctcc atagaagaca ccgggaccga tccagcctcc gcggccccga attcgagctc
     2401 ggtacccggg gatcctctag tcagctgacg cgtatggccg tcaatgtgta ctccacatct
     2461 gtgaccagtg aaaatctgag tcgccatgat atgcttgcat gggtcaacga ctccctgcac
     2521 ctcaactata ccaagataga acagctttgt tcaggggcag cctactgcca gttcatggac
     2581 atgctcttcc ccggctgtgt gcacttgagg aaagtgaagt tccaggccaa actagagcat
     2641 gaatacatcc acaacttcaa ggtgctgcaa gcagctttca agaagatggg tgttgacaaa
     2701 atcattcctg tagagaaatt agtgaaagga aaattccaag ataattttga gtttattcag
     2761 tggtttaaga aattctttga cgcaaactat gatggaaagg attacaaccc tctgctggcg
     2821 cggcagggcc aggacgtagc gccacctcct aacccaggtg atcagatctt caacaaatcc
     2881 aagaaactca ttggcacagc agttccacag aggacgtccc ccacaggccc aaaaaacatg
     2941 cagacctctg gccggctgag caatgtggcc cccccctgca ttctccggaa gaatcctcca
     3001 tcatctagag gtggcggggg cagtggggga tcccatactc tttatgcccc cggtggatat
     3061 gacattatgg gatatctgtg ccagatcatg aaccggccaa acccgcaggt ggaactgggc
     3121 cccgtggata catcctgcgc cttggttctt tgtgacctga aacagaaaga caccccggta
     3181 gtttacgcga gtgaagcctt cctctacatg acaggttaca gcaacgcaga ggtgctgggc
     3241 cggaattgcc ggtttctgca aagccctgac ggcatggtga agcccaagag cacccggaag
     3301 tacgtggata gtaacacaat caatactatg cgcaaggcaa tcgacaggaa tgccgaggtg
     3361 caggttgaag tagtcaattt taaaaagaat ggacagcgat ttgttaattt cctgactatg
     3421 atacctgtta gggacgaaac aggagagtat cgatactcta tgggattcca gtgcgaagtc
     3481 gacgggagcg ggggtgggag taagcttatg gtgtctaagg gcgaggagct gttcaccggg
     3541 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtccgc
     3601 ggcgagggcg agggcgatgc caccaacggc aagctgaccc tgaagttcat ctgcaccacc
     3661 ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cgtggcctgc
     3721 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     3781 ggctacgtcc aggagcgcac catctctttc aaggacgacg gtacctacaa gacccgcgcc
     3841 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     3901 aaggaggacg gcaacatcct ggggcacaag ctggagtaca acttcaacag ccactacgtc
     3961 tatatcacgg ccgacaagca gaagaactgc atcaaggcta acttcaagat ccgccacaac
     4021 gttgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     4081 ggccccgtgc tgctgcccga caaccactac ctgagccatc agtccaagct gagcaaagac
     4141 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggattaca
     4201 catggcatgg acgagctgta caaggctagc ggaggcggtg gctcaggcgg aggagggtca
     4261 ggagggggag gcagtggcgg gggcggctct ggaggagggg gttctggtgg tggtagctac
     4321 gtactggcaa ccacactgga acggatcgag aaaaatttcg tgattactga tccgagactg
     4381 cctgacaacc caatcatttt tgcgagcgat tccttcctgc agctgacaga atattctcgg
     4441 gaagagatcc tggggcgcaa ttgccgtttt ctgcagggac ccgagacaga ccgtgccact
     4501 gttcggaaaa tcagagatgc tattgacaac cagactgaag tgaccgttca gctgatcaat
     4561 tataccaaga gcggcaagaa gttctggaac gtgttccacc tgcagccgat gcgcgattat
     4621 aagggcgacg tccagtactt cattggcgtg cagctggatg gcaccgaacg tcttcatggc
     4681 gccgctgagc gtgaggcggt ctgcctgatc aaaaagacag cctttcagat tgctgaggca
     4741 gcgaacgacg aaaattactt ttgatagtct agacgcgtgc tagcgcggcc gcatcgataa
     4801 gcttgtcgac gatatcgaat tccacggggt tggggttgcg ccttttccaa ggcagccctg
     4861 ggtttgcgca gggacgcggc tgctctgggc gtggttccgg gaaacgcagc ggcgccgacc
     4921 ctgggtctcg cacattcttc acgtccgttc gcagcgtcac ccggatcttc gccgctaccc
     4981 ttgtgggccc cccggcgacg cttcctgctc cgcccctaag tcgggaaggt tccttgcggt
     5041 tcgcggcgtg ccggacgtga caaacggaag ccgcacgtct cactagtacc ctcgcagacg
     5101 gacagcgcca gggagcaatg gcagcgcgcc gaccgcgatg ggctgtggcc aatagcggct
     5161 gctcagcagg gcgcgccgag agcagcggcc gggaaggggc ggtgcgggag gcggggtgtg
     5221 gggcggtagt gtgggccctg ttcctgcccg cgcggtgttc cgcattctgc aagcctccgg
     5281 agcgcacgtc ggcagtcggc tccctcgttg accgaatcac cgacctctct ccccaggggg
     5341 atccgtttat ctgcagaatt cgcccttgac gtcatgtcta gactggacaa gagcaaagtc
     5401 ataaacggcg ctctggaatt actcaatgga gtcggtatcg aaggcctgac gacaaggaaa
     5461 ctcgctcaaa agctgggagt tgagcagcct accctgtact ggcacgtgaa gaacaagcgg
     5521 gccctgctcg atgccctgcc aatcgagatg ctggacaggc atcataccca cttctgcccc
     5581 ctggaaggcg agtcatggca agactttctg cggaacaacg ccaagtcatt ccgctgtgct
     5641 ctcctctcac atcgcgacgg ggctaaagtg catctcggca cccgcccaac agagaaacag
     5701 tacgaaaccc tggaaaatca gctcgcgttc ctgtgtcagc aaggcttctc cctggagaac
     5761 gcactgtacg ctctgtccgc cgtgggccac tttacactgg gctgcgtatt ggaggaacag
     5821 gagcatcaag tagcaaaaga ggaaagagag acacctacca ccgattctat gcccccactt
     5881 ctgagacaag caattgagct gttcgaccgg cagggagccg aacctgcctt ccttttcggc
     5941 ctggaactaa tcatatgtgg cctggagaaa cagctaaagt gcgaaagcgg cgggccggcc
     6001 gacgcccttg acgattttga cttagacatg ctcccagccg atgcccttga cgactttgac
     6061 cttgatatgc tgcctgctga cgctcttgac gattttgacc ttgacatgct ccccgggtaa
     6121 ggatcccgcc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
     6181 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     6241 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     6301 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     6361 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     6421 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     6481 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     6541 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     6601 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     6661 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aagcttgcca caacccacaa
     6721 ggagacgacc ttccatgaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga
     6781 tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg
     6841 ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg
     6901 gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg
     6961 aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac
     7021 agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg
     7081 cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat
     7141 tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg
     7201 atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc
     7261 aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc
     7321 acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg
     7381 actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga
     7441 ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc
     7501 ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc
     7561 agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa
     7621 tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg
     7681 tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc
     7741 gtccgagggc aaaggaatag ctgcagcacg tgtctagaaa cgctggtcga caatcaacct
     7801 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     7861 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     7921 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     7981 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     8041 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     8101 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     8161 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     8221 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     8281 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     8341 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattcg agctcggtac
     8401 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     8461 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     8521 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     8581 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     8641 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     8701 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     8761 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     8821 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     8881 tcaatgtatc ttatcatgtc tggctcyrsc yatcccgccc ctaactccgc ccagttccgc
     8941 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     9001 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc
     9061 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
     9121 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     9181 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     9241 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     9301 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     9361 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     9421 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     9481 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     9541 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     9601 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     9661 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     9721 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     9781 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     9841 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     9901 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     9961 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
    10021 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
    10081 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
    10141 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
    10201 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
    10261 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
    10321 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
    10381 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
    10441 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
    10501 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
    10561 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
    10621 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
    10681 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
    10741 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
    10801 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
    10861 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
    10921 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
    10981 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
    11041 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
    11101 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
    11161 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
    11221 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
    11281 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
    11341 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
    11401 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
    11461 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
    11521 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
    11581 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
    11641 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
    11701 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
    11761 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
    11821 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
    11881 caattaaccc tcactaaagg gaacaaaagc tggagctgca agctt
//
