LOCUS       sequence-477256-       14649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac plasmid for inducible PEmax expression and Puromycin 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14649)
  AUTHORS   Chen W, Choi J, Li X, Nathans JF, Martin B, Yang W, Hamazaki N, Qiu 
            C, Lalanne JB, Regalado S, Kim H, Agarwal V, Nichols E, Leith A, Lee
            C, Shendure J
  TITLE     Symbolic recording of signalling and cis-regulatory element activity
            to DNA.
  JOURNAL   Nature. 2024 Aug;632(8027):1073-1081. doi: 
            10.1038/s41586-024-07706-4. Epub 2024 Jul 17.
  PUBMED    39020177
REFERENCE   2  (bases 1 to 14649)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41586-024-07706-4"; journalName: "Nature"; date: 
            "2024-08"; volume: "632"; issue: "8027"; pages: "1073-1081"
COMMENT     Sequence Label: P022-mPB-TRE-PEmax-T2A-mCherry-PGK-rttA-Puro
FEATURES             Location/Qualifiers
     source          1..14649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..67
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        483..797
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    491..509
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    527..545
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    562..580
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    598..616
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    634..652
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    669..687
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    705..723
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     770..794
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             855..875
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             876..4976
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRKLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLKREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5034..5054
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7155..7175
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7185..7211
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7221..7274
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             7275..7985
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(7420..7438)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(8007..8024)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8013..8237
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        8291..8801
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     8720..8738
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             join(8818..9435,9436..9441,9442..9477,9478..9480,9481..9516,
                     9517..9519,9520..9555,9556..9561)
                     /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
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             9571..9627
                     /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             9628..10227
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(9628..9647)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     10124..10144
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(10288..10308)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    10350..10471
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10387..10406)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10441..10460
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   11234..11268
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(11759..11775)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11759..11775)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11772..11794)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11783..11799
                     /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(11807..11813,11814..11831,11832..11837))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11852..11873
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11990..12007)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12161..12749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12241..12260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12920..13711,13712..13780))
                     /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     13543..13562
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13781..13885)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(13911..14366)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14048..14069)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     14260..14279
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14493..14515
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     14507..14524
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     14508..14524
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttaaccctag aaagataatc atattgtgac gtacgttaaa gataatcatg tgtaaaattg
       61 acgcatgtgt tttatcggtc tgtatatcga ggtttattta ttaatttgaa tagatattaa
      121 gttttattat atttacactt acatactaat aataaattca acaaacaatt tatttatgtt
      181 tatttattta ttaaaaaaaa caaaaactca aaatttcttc tataaagtaa caaaactttt
      241 atgagggaca gccccccccc aaagccccca gggatgtaat tacgtccctc ccccgctagg
      301 gggcagcagc gagccgcccg gggctccgct ccggtccggc gctccccccg catccccgag
      361 ccggcagcgt gcggggacag cccgggcacg gggaaggtgg cacgggatcg ctttcctctg
      421 aacgcttctc gctgctcttt gagcctgcag acacctgggg ggatacgggg aaaaaccggt
      481 gggagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
      541 agagaacgat gtcgagttta ctccctatca gtgatagaga acgtatgtcg agtttactcc
      601 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
      661 cgagtttatc cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag
      721 agaacgtatg tcgaggtagg cgtgtacggt gggaggccta tataagcaga gctcgtttag
      781 tgaaccgtca gatcgccgat cccacgaagt ccgccaccat gaaacggaca gccgacggaa
      841 gcgagttcga gtcaccaaag aagaagcgga aagtcgacaa gaagtacagc atcggcctgg
      901 acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca
      961 agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag
     1021 ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa
     1081 gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga
     1141 tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg
     1201 ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg
     1261 agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg
     1321 acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga
     1381 tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc
     1441 agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg
     1501 ccatcctgtc tgccagactg agcaagagca gaaagctgga aaatctgatc gcccagctgc
     1561 ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc
     1621 ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca
     1681 cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt
     1741 ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca
     1801 ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc
     1861 aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga
     1921 ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg
     1981 aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc
     2041 tcgtgaagct gaagagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca
     2101 tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt
     2161 acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct
     2221 actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg
     2281 aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga
     2341 gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca
     2401 agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg
     2461 tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg
     2521 acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca
     2581 agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct
     2641 ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg
     2701 aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag
     2761 agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc
     2821 agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca
     2881 tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca
     2941 acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga
     3001 aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca
     3061 gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag
     3121 tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca
     3181 cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag
     3241 agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga
     3301 agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca
     3361 tcaaccggct gtccgactac gatgtggacg ctatcgtgcc tcagagcttt ctgaaggacg
     3421 actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg
     3481 tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca
     3541 agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg
     3601 aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc
     3661 acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga
     3721 tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt
     3781 tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga
     3841 acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt
     3901 acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg
     3961 gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga
     4021 ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg
     4081 gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc
     4141 cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta
     4201 tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga
     4261 agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg
     4321 aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg
     4381 aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag
     4441 tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc
     4501 ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct
     4561 ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg
     4621 aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca
     4681 tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag
     4741 tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca
     4801 tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca
     4861 ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc
     4921 agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga ggtgactccg
     4981 gcggaagctc tggtggcagc aagcggaccg ccgacggctc tgaattcgag agccctaaga
     5041 agaaaagaaa ggtgagcgga ggctctagcg gcggaagcac cctgaacatt gaagacgagt
     5101 atagactgca tgaaacaagc aaggaacccg acgtgtccct gggctccacc tggctgtccg
     5161 actttcccca ggcctgggcc gagacaggag gaatgggcct ggccgtgcgg caggcacccc
     5221 tgatcatccc tctgaaggcc acctctacac ccgtgagcat caagcagtac cctatgtctc
     5281 aggaggccag actgggcatc aagcctcaca tccagaggct gctggaccag ggcatcctgg
     5341 tgccatgcca gagcccctgg aacacaccac tgctgcccgt gaagaagcca ggcaccaatg
     5401 actatagacc cgtgcaggat ctgagagagg tgaacaagag ggtggaggat atccacccca
     5461 ccgtgcccaa cccttacaat ctgctgtccg gcctgccccc ttctcaccag tggtatacag
     5521 tgctggacct gaaggatgcc ttcttttgtc tgagactgca ccctaccagc cagccactgt
     5581 tcgcctttga gtggagggac cctgagatgg gcatctctgg ccagctgacc tggacacgcc
     5641 tgcctcaggg cttcaagaat agcccaacac tgtttaacga ggccctgcac cgcgacctgg
     5701 cagatttccg gatccagcac ccagatctga tcctgctgca gtacgtggac gatctgctgc
     5761 tggccgccac cagcgagctg gattgccagc agggaacacg cgccctgctg cagaccctgg
     5821 gaaacctggg atatagggca tccgccaaga aggcccagat ctgtcagaag caggtgaagt
     5881 acctgggcta tctgctgaag gagggccaga gatggctgac agaggccagg aaggagacag
     5941 tgatgggcca gccaacaccc aagaccccaa gacagctgag ggagttcctg ggcaaagcag
     6001 gattttgcag gctgttcatc ccaggattcg cagagatggc agcacctctg tacccactga
     6061 ccaagccggg caccctgttt aattggggcc ctgaccagca gaaggcctat caggagatca
     6121 agcaggccct gctgacagca ccagccctgg gcctgccaga cctgaccaag cctttcgagc
     6181 tgtttgtgga tgagaagcag ggctacgcca agggcgtgct gacccagaag ctgggaccat
     6241 ggcgacggcc cgtggcctat ctgtccaaga agctggaccc agtggcagca ggatggccac
     6301 catgcctgag gatggtggca gcaatcgccg tgctgacaaa ggatgccggc aagctgacca
     6361 tgggacagcc actggtcatc ctggcaccac acgcagtgga ggccctggtg aagcagcctc
     6421 cagatcgctg gctgtctaac gcccggatga cacactacca ggccctgctg ctggacaccg
     6481 atcgcgtgca gtttggccct gtggtggccc tgaatccagc caccctgctg cctctgccag
     6541 aggagggcct gcagcacaac tgtctggaca tcctggcaga ggcacacgga acaaggccag
     6601 acctgaccga tcagcccctg cctgacgccg atcacacatg gtataccgat ggaagctccc
     6661 tgctgcagga gggccagagg aaggcaggag cagcagtgac cacagagaca gaagtgatct
     6721 gggccaaggc cctgccagca ggcacatccg cccagcgggc cgagctgatc gccctgaccc
     6781 aggccctgaa gatggccgag ggcaagaagc tgaacgtgta cacagactcc agatatgcct
     6841 tcgccaccgc acacatccac ggagagatct acaggcgccg gggctggctg acctctgagg
     6901 gcaaggagat caagaacaag gatgagatcc tggccctgct gaaggccctg tttctgccca
     6961 agcggctgag catcatccac tgtcctggac accagaaggg acactccgcc gaggcaaggg
     7021 gcaatcggat ggccgaccag gccgccagaa aggctgctat tactgaaact cccgacactt
     7081 ccactctgct gattgaaaac tcctcccctt ctggcggctc aaaaagaacc gccgacggca
     7141 gcgaattcga gtctcccaag aagaagagga aagtcggctc tggccctgcc gctaagagag
     7201 tgaagctgga cggatctgga gagggaaggg gaagcctcct aacttgcgga gatgtcgagg
     7261 agaatcctgg cccaatggtg agcaagggcg aggaggataa catggccatc atcaaggagt
     7321 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     7381 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     7441 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     7501 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     7561 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     7621 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     7681 ccgacggccc cgtaatgcag aagaaaacca tgggctggga ggcctcctcc gagcggatgt
     7741 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     7801 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     7861 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     7921 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     7981 agtaacgaaa cgcggccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     8041 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     8101 ctaataaaat gagaaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     8161 tggggtgggg caggacagca agggggagga ttgggaacac aatagcaggc atgctgggga
     8221 tgcggtgggc tctatgggga tcgctttcct ctgaacgctt ctcgctgctc tttgagccat
     8281 ggcagacacc ggggttgggg ttgcgccttt tccaaggcag ccctgggttt gcgcagggac
     8341 gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg tcacgcacat
     8401 tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg ggccccccgg
     8461 cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga
     8521 cgtgacaaac ggaagccgca cctctcacta gtaccctcgc agacggacag cgccagggag
     8581 caatggcagc gcgccgaccg cgatgggctg tggccaatag cggctgctca gcagggcgcg
     8641 ccgagagcag cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg
     8701 ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc acgtcggcag
     8761 tcggctccct cgttgaccga atcaccgacc tctctcccca ggctagccac cgccaccatg
     8821 agcagactag acaagagcaa agtcataaac ggagctctgg aattactcaa tggtgtcggt
     8881 atcgaaggcc tgacgacaag gaaactcgct caaaagctgg gagttgagca gcctaccctg
     8941 tactggcacg tgaagaacaa gcgggccctg ctcgatgccc tgccaatcga gatgctggac
     9001 aggcatcata cccacttctg ccccctggaa ggcgagtcat ggcaagactt tctgcggaac
     9061 aacgccaagt cataccgctg tgctctcctc tcacatcgcg acggggctaa agtgcatctc
     9121 ggcacccgcc caacagagaa acagtacgaa accctggaaa atcagctcgc gttcctgtgt
     9181 cagcaaggct tctccctgga gaacgcactg tacgctctgt ccgccgtggg ccactttaca
     9241 ctgggctgcg tattggagga acaggagcat caagtagcaa aagaggaaag agagacacct
     9301 accaccgatt ctatgccccc acttctgaga caagcaattg agctgttcga ccggcaggga
     9361 gccgaacctg ccttcctttt cggcctggaa ctaatcatat gtggcctgga gaaacagcta
     9421 aagtgcgaaa gcggcgggcc gaccgacgcc cttgacgatt ttgacttaga catgctccca
     9481 gccgatgccc ttgacgactt tgaccttgat atgctgcctg ctgacgctct tgacgatttt
     9541 gaccttgaca tgctccccgg gggatctgga gctactaact tcagcctgct gaagcaggct
     9601 ggcgacgtgg aggagaaccc tggacccatg accgagtaca agcccacggt gcgcctcgcc
     9661 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     9721 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     9781 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     9841 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     9901 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     9961 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggtgtctcg
    10021 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
    10081 gagcgcgccg gggtgcccgc cttcctggag acatccgcgc cccgcaacct ccccttctac
    10141 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
    10201 tgcatgaccc gcaagcccgg tgcctaacgt cgacaatcaa cctctggatt acaaaatttg
    10261 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
    10321 tttaatgcct ttgtatcatg cgttaactaa acttgtttat tgcagcttat aatggttaca
    10381 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
    10441 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gaattgactc aaatgatgtc
    10501 aattagtcta tcagaagctc atctggactc ccttccgggg gacaagacat ccctgtttaa
    10561 tatttaaaca gcagtgttcc caaactgggt tcttatatcc cttgctctgg tcaaccaggt
    10621 tgcagggttt cctgtcctca caggaacgaa gtccctaaag aaacagtggc agccaggttt
    10681 agccccggaa ttgactggat tcctttttta gggcccattg gtatggcttt ttccccgtat
    10741 ccccccaggt gtctgcaggc tcaaagagca gcgagaagcg ttcagaggaa agcgatcccg
    10801 tgccaccttc cccgtgcccg ggctgtcccc gcacgctgcc ggctcgggga tgcgggggga
    10861 gcgccggacc ggagcggagc cccgggcggc tcgctgctgc cccctagcgg gggagggacg
    10921 taattacatc cctgggggct ttgggggggg gctgtccctg atatctataa caagaaaata
    10981 tatatataat aagttatcac gtaagtagaa catgaaataa caatataatt atcgtatgag
    11041 ttaaatctta aaagtcacgt aaaagataat catgcgtcat tttgactcac gcggtcgtta
    11101 tagttcaaaa tcagtgacac ttaccgcatt gacaagcacg cctcacggga gctccaagcg
    11161 gcgactgaga tgtcctaaat gcacagcgac ggattcgcgc tatttagaaa gagagagcaa
    11221 tatttcaaga atgcatgcgt caattttacg cagactatct ttctagggtt aatctagctg
    11281 catcaggatc atatcgtcgg gtcttttttc cggctcagtc atcgcccaag ctggcgctat
    11341 ctgggcatcg gggaggaaga agcccgtgcc ttttcccgcg aggttgaagc ggcatggaaa
    11401 gagtttgccg aggatgactg ctgctgcatt gacgttgagc gaaaacgcac gtttaccatg
    11461 atgattcggg aaggtgtggc catgcacgcc tttaacggtg aactgttcgt tcaggccacc
    11521 tgggatacca gttcgtcgcg gcttttccgg acacagttcc ggatggtcag cccgaagcgc
    11581 atcagcaacc cgaacaatac cggcgacagc cggaactgcc gtgccggtgt gcagattaat
    11641 gacagcggtg cggcgctggg atattacgtc agcgaggacg ggtatcctgg ctggatgccg
    11701 cagaaatgga catggatacc ccgtgagtta cccggcgggc gcgcttggcg taatcatggt
    11761 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
    11821 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
    11881 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
    11941 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
    12001 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
    12061 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
    12121 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
    12181 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
    12241 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
    12301 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
    12361 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
    12421 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
    12481 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
    12541 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
    12601 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
    12661 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
    12721 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
    12781 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
    12841 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
    12901 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
    12961 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
    13021 agggcttacc atctggcccc agtgctgcaa tgataccgcg agatccacgc tcaccggctc
    13081 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
    13141 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
    13201 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
    13261 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
    13321 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
    13381 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
    13441 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
    13501 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
    13561 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
    13621 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
    13681 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
    13741 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
    13801 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
    13861 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag
    13921 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
    13981 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
    14041 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
    14101 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
    14161 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
    14221 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
    14281 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
    14341 gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga
    14401 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
    14461 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
    14521 cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg gaggacgggc agactcgcgg
    14581 tgcaaatgtg ttttacagcg tgatggagca gatgaagatg ctcgacacgc tgcagaacac
    14641 gcagctaga
//
