LOCUS       sequence-373504-       11776 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FKBP12 (F36V mutant) degron-tagged dCas9 fused with tagRFPt, P2A 
            site and tagBFP, expressed under EF1-alpha promoter in a piggyBac 
            plasmid. Expresses Blasticidin resistance under PGK promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11776)
  AUTHORS   Noviello G, Gjaltema RAF, Schulz EG
  TITLE     CasTuner is a degron and CRISPR/Cas-based toolkit for analog tuning 
            of endogenous gene expression.
  JOURNAL   Nat Commun. 2023 Jun 3;14(1):3225. doi: 10.1038/s41467-023-38909-4.
  PUBMED    37270532
REFERENCE   2  (bases 1 to 11776)
  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: "Nat 
            Commun."; date: "2023-06-3"; volume: "14(1):3225. doi"; pages: " 
            10.1038/s41467-023-38909-4"
COMMENT     Sequence Label: 
            pSLPB2B-FKBP12_F36V-SpdCas9-tagRFPt-P2A-tagBFP-PGK-Blasticidin
FEATURES             Location/Qualifiers
     source          1..11776
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     412..431
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(650..754)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(780..1235)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(917..938)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1129..1148
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1362..1384
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1376..1393
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1377..1393
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1403..1422
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1403..1421
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   1437..1471
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1793..2004
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2017..2285
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2337..2357
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2367..2687
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             join(2739..2762,2763..2765,2766..5252,5253..5255,
                     5256..6839)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             2739..6839
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6840..6860
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6861..6878
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6933..7661
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="VSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMR
                     IKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
                     LTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTDM
                     ALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVA
                     RYCDLPSKLGHKLNGMDELYK"
     CDS             7680..7736
                     /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             7737..8438
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    8453..8677
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     9028..9047
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             9103..9501
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9502..10090
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9555..9575)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9973..9984)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     repeat_region   10273..10335
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(10365..10385)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(10365..10383)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10404..10420)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10404..10420)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10417..10439)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10428..10444
                     /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(10452..10458,10459..10476,10477..10482))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10497..10518
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10635..10652)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10806..11394)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10886..10905)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11565..11776,1..580,581..649))
                     /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"
ORIGIN
        1 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
       61 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
      121 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
      181 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
      241 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
      301 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
      361 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
      421 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
      481 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
      541 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
      601 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
      661 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
      721 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
      781 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
      841 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
      901 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
      961 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     1021 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     1081 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     1141 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     1201 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     1261 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     1321 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     1381 aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg ggctaaccct
     1441 agaaagatag tctgcgtaaa attgacgcat gcattcttga aatattgctc tctctttcta
     1501 aatagcgcga atccgtcgct gtgcatttag gacatctcag tcgccgcttg gagctcccgt
     1561 gaggcgtgct tgtcaatgcg gtaagtgtca ctgattttga actataacga ccgcgtgagt
     1621 caaaatgacg catgattatc ttttacgtga cttttaagat ttaactcata cgataattat
     1681 attgttattt catgttctac ttacgtgata acttattata tatatatttt cttgttatag
     1741 atatcaacta gaatacgcgt aaggatctgc gatcgctccg gtgcccgtca gtgggcagag
     1801 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc
     1861 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
     1921 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
     1981 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc
     2041 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc
     2101 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga
     2161 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt
     2221 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac
     2281 agatccaagc tgtgaccggc gcctactcta gagctagcga attcgccgcc accatgccta
     2341 agaaaaagag gaaggtgagc ggccgcggag ttcaggtgga aaccatctcc ccaggagacg
     2401 ggcgcacctt ccccaagcgc ggccagacct gcgtggtgca ctacaccggg atgcttgaag
     2461 atggaaagaa agttgattcc tcccgggaca gaaacaagcc ctttaagttt atgctaggca
     2521 agcaggaggt gatccgaggc tgggaagaag gggttgccca gatgagtgtg ggtcagagag
     2581 ccaaactgac tatatctcca gattatgcct atggtgccac tgggcaccca ggcatcatcc
     2641 caccacatgc cactctcgtc ttcgatgtgg agcttctaaa actggaagga tccggcggat
     2701 ctggaggtag tggtggatct ggaggttctg gtggctccga caagaagtac agcatcggcc
     2761 tggccatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
     2821 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
     2881 gcgccctgct gttcgacagc ggagaaacag ccgaggccac ccggctgaag agaaccgcca
     2941 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
     3001 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
     3061 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
     3121 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
     3181 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
     3241 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
     3301 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
     3361 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc
     3421 tgcccggcga gaagaagaat ggcctgttcg gcaacctgat tgccctgagc ctgggcctga
     3481 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg
     3541 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
     3601 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
     3661 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc
     3721 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
     3781 agattttctt cgaccagagc aagaacggct acgccggcta catcgatggc ggagccagcc
     3841 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac
     3901 tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca
     3961 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt
     4021 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc
     4081 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga
     4141 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gccagcgccc
     4201 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc
     4261 ccaagcacag cctgctgtac gagtacttca ccgtgtacaa cgagctgacc aaagtgaaat
     4321 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaagccatcg
     4381 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
     4441 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
     4501 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
     4561 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
     4621 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
     4681 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
     4741 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
     4801 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
     4861 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
     4921 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
     4981 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
     5041 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
     5101 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
     5161 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
     5221 acatcaaccg gctgtccgac tacgatgtgg acgctatcgt gcctcagagc tttctgaagg
     5281 acgactccat cgataacaaa gtgctgactc ggagcgacaa gaaccggggc aagagcgaca
     5341 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcgccag ctgctgaatg
     5401 ccaagctgat tacccagagg aagttcgaca atctgaccaa ggccgagaga ggcggcctga
     5461 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
     5521 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aacgacaaac
     5581 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
     5641 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
     5701 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg
     5761 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
     5821 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
     5881 agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa
     5941 caggcgagat cgtgtgggat aagggccggg actttgccac cgtgcggaaa gtgctgtcta
     6001 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt
     6061 ctatcctgcc caagaggaac agcgacaagc tgatcgccag aaagaaggac tgggacccta
     6121 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag
     6181 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca
     6241 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag
     6301 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg
     6361 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc
     6421 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc
     6481 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caaacactac ctggacgaga
     6541 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca
     6601 aggtgctgag cgcctacaac aagcacagag acaagcctat cagagagcag gccgagaata
     6661 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca
     6721 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc
     6781 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgacc
     6841 ccaagaagaa gcgcaaggtg caccaccacc accaccacgg aggaggtgga agcggaggag
     6901 gaggaagcgg aggaggaggt agctggcgcg ccgtgtctaa gggcgaagag ctgattaagg
     6961 agaacatgca catgaagctg tacatggagg gcaccgtgaa caaccaccac ttcaagtgca
     7021 catccgaggg cgaaggcaag ccctacgagg gcacccagac catgagaatc aaggtggtcg
     7081 agggcggccc tctccccttc gccttcgaca tcctggctac cagcttcatg tacggcagca
     7141 gaaccttcat caaccacacc cagggcatcc ccgacttctt taagcagtcc ttccctgagg
     7201 gcttcacatg ggagagagtc accacatacg aagacggggg cgtgctgacc gctacccagg
     7261 acaccagcct ccaggacggc tgcctcatct acaacgtcaa gatcagaggg gtgaacttcc
     7321 catccaacgg ccctgtgatg cagaagaaaa cactcggctg ggaggccaac accgagatgc
     7381 tgtaccccgc tgacggcggc ctggaaggca gaaccgacat ggccctgaag ctcgtgggcg
     7441 ggggccacct gatctgcaac ttcaagacca catacagatc caagaaaccc gctaagaacc
     7501 tcaagatgcc cggcgtctac tatgtggacc acagactgga aagaatcaag gaggccgaca
     7561 aagagaccta cgtcgagcag cacgaggtgg ctgtggccag atactgcgac ctccctagca
     7621 aactggggca caaacttaat ggcatggacg agctgtacaa gttaattaac ggaagcggag
     7681 ctactaactt cagcctgctg aagcaggctg gagacgtgga ggagaaccct ggacctatga
     7741 gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc gtggacaacc
     7801 atcacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga
     7861 gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctactagct
     7921 tcctctacgg cagcaagacc ttcatcaacc acacccaggg catccccgac ttcttcaagc
     7981 agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc
     8041 tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac gtcaagatca
     8101 gaggggtgaa cttcacatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg
     8161 ccttcaccga gacgctgtac cccgctgacg gcggcctgga aggcagaaac gacatggccc
     8221 tgaagctcgt gggcgggagc catctgatcg caaacatcaa gaccacatat agatccaaga
     8281 aacccgctaa gaacctcaag atgcctggcg tctactatgt ggactacaga ctggaaagaa
     8341 tcaaggaggc caacaacgag acctacgtcg agcagcacga ggtggcagtg gccagatact
     8401 gcgacctccc tagcaaactg gggcacaagc ttaattagac gcgtattaac gactgtgcct
     8461 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     8521 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     8581 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     8641 aatagcaggc atgctgggga tgcggtgggc tctatggcct ctggcctcgc acacattcca
     8701 catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta
     8761 ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac
     8821 aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga
     8881 agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc
     8941 agaggctggg aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg
     9001 gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg
     9061 cgctgttctc ctcttcctca tctccgggcc tttcgacctg caatggccaa gcctttgtct
     9121 caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct
     9181 gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc
     9241 aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct
     9301 gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc
     9361 ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc
     9421 atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct
     9481 ggttatgtgt gggagggcta aaatcaacct ctggattaca aaatttgtga aagattgact
     9541 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     9601 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     9661 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     9721 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     9781 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     9841 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     9901 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     9961 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
    10021 ctgcggcctc ttccgcgtct ccgccttcgc cctcagacga gtcggatctc cctttgggcc
    10081 gcctccccgc cggtacctaa aagttttgtt actttataga agaaattttg agtttttgtt
    10141 tttttttaat aaataaataa acataaataa attgtttgtt gaatttatta ttagtatgta
    10201 agtgtaaata taataaaact taatatctat tcaaattaat aaataaacct cgatatacag
    10261 accgataaaa cacatgcgtc aattttacgc atgattatct ttaacgtacg tcacaatatg
    10321 attatctttc tagggttaat ctagtattaa tccagctttt gttcccttta gtgagggtta
    10381 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
    10441 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
    10501 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
    10561 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
    10621 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
    10681 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
    10741 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
    10801 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
    10861 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
    10921 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
    10981 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
    11041 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
    11101 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
    11161 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
    11221 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
    11281 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
    11341 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
    11401 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
    11461 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
    11521 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
    11581 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
    11641 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
    11701 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
    11761 gccgagcgca gaagtg
//
