LOCUS       sequence-331354-       14593 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac vector to express GA-inducible VPR-Sa_dCas9 with hygromycin
            resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14593)
  AUTHORS   Wang S, Matsumoto K, Lish SR, Cartagena-Rivera AX, Yamada KM
  TITLE     Budding epithelial morphogenesis driven by cell-matrix versus 
            cell-cell adhesion.
  JOURNAL   Cell. 2021 Jul 8;184(14):3702-3716.e30. doi: 
            10.1016/j.cell.2021.05.015. Epub 2021 Jun 15.
  PUBMED    34133940
REFERENCE   2  (bases 1 to 14593)
  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: 
            "10.1016/j.cell.2021.05.015"; journalName: "Cell"; date: "2021-07-8-
            8"; volume: "184"; issue: "14"; pages: "3702-3716.e30"
COMMENT     Sequence Label: pW210-pPB-GA-inducible-CRISPRa-sa_dCas9-VPR-hygR
FEATURES             Location/Qualifiers
     source          1..14593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..284
                     /label=chicken beta-actin promoter
     intron          286..1303
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1227..1249
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1311..1330
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1358..1367
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2426..2575
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             2582..2602
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3032..3403
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     CDS             3422..3991
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     misc_feature    join(4009..4582,4583..4585,4586..4594)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4176..4193)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4403..4422
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4625..5644
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             4625..5644
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     misc_feature    5682..6270
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5735..5755)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6153..6164)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        6291..6790
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6326..6349)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6723..6742
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6832..6849
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             7123..7821
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             7843..7863
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11044..11091
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     repeat_region   11281..11343
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(11394..11414)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(11394..11412)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11433..11449)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11433..11449)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11446..11468)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11457..11473
                     /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(11481..11487,11488..11505,11506..11511))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11526..11547
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11664..11681)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11835..12423)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11915..11934)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12594..13385,13386..13454))
                     /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     13217..13236
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13455..13559)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(13585..14040)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(13722..13743)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     13934..13953
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14167..14189
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     14181..14198
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     14182..14198
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     14208..14227
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        14208..14226
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   14256..14290
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
       61 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg
      121 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
      181 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
      241 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcggggagt cgctgcgacg
      301 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
      361 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta
      421 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct
      481 ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt
      541 ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc
      601 ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg
      661 gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg
      721 agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt
      781 tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc
      841 cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc
      901 cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc
      961 ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt
     1021 gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg
     1081 cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc
     1141 gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg
     1201 ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag
     1261 agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct
     1321 ggttattgtg ctgtctcatc attttggcaa agaattcgcc accatggctg cgagcgatga
     1381 agttaatctt attgagagca gaacagtggt tcctctcaat acatgggttt taatatccaa
     1441 cttcaaagta gcctacaata tccttcgtcg ccctgatgga acctttaacc gacacttagc
     1501 tgagtatcta gaccgtaaag tcactgcaaa cgccaatccg gttgatgggg ttttctcgtt
     1561 cgatgtcttg attgatcgca ggatcaatct tctaagcaga gtctatagac cagcttatgc
     1621 agatcaagag caacctccta gtattttaga tctcgagaag cctgttgatg gcgacattgt
     1681 ccctgttata ttgttcttcc atggaggtag ctttgctcat tcttctgcaa acagtgccat
     1741 ctacgatact ctttgtcgca ggcttgttgg tttgtgcaag tgtgttgttg tctctgtgaa
     1801 ttatcggcgt gcaccagaga atccataccc ttgtgcttat gatgatggtt ggattgctct
     1861 taattgggtt aactcgagat cttggcttaa atccaagaaa gactcaaagg tccatatttt
     1921 cttggctggt gatagctctg gaggtaacat cgcgcataat gtggctttaa gagcgggtga
     1981 atcgggaatc gatgttttgg ggaacattct gctgaatcct atgtttggtg ggaatgagag
     2041 aacggagtct gagaaaagtt tggatgggaa atactttgtg acggttagag accgcgattg
     2101 gtactggaaa gcgtttttac ccgagggaga agatagagag catccagcgt gtaatccgtt
     2161 tagcccgaga gggaaaagct tagaaggagt gagtttcccc aagagtcttg tggttgtcgc
     2221 gggtttggat ttgattagag attggcagtt ggcatacgcg gaagggctca agaaagcggg
     2281 tcaagaggtt aagcttatgc atttagagaa agcaactgtt gggttttacc tcttgcctaa
     2341 taacaatcat ttccataatg ttatggatga gatttcggcg tttgtaaacg cggaatgtga
     2401 attctcaagc gctgcaggta ctagtgatgc tttagacgat tttgacttag atatgcttgg
     2461 ttcagacgcg ttagacgact tcgacctaga catgttaggc tcagatgcat tggacgactt
     2521 cgatttagat atgttgggct ccgatgccct agatgacttt gatctagata tgctaggtag
     2581 tcccaaaaag aagaggaaag tgggatccca gtatctgccc gacacagatg atagacaccg
     2641 aatcgaagag aaacgcaagc gaacgtatga aaccttcaaa tcgatcatga agaaatcgcc
     2701 cttctcgggt ccgaccgatc ccaggccccc accgagaagg attgcggtcc cgtcccgctc
     2761 gtcggccagc gtgccgaagc ctgcgccgca gccctacccc ttcacgtcga gcctgagcac
     2821 aatcaattat gacgagttcc cgacgatggt gttcccctcg ggacaaatct cacaagcctc
     2881 ggcgctcgca ccagcgcctc cccaagtcct tccgcaagcg cctgccccag cgcctgcacc
     2941 ggcaatggtg tccgccctcg cacaggcccc tgcgcccgtc cccgtgctcg cgcctggacc
     3001 gccccaggcg gtcgctccac cggctccgaa gccgacgcag gccggagagg gaacactctc
     3061 cgaagcactt cttcaactcc agtttgatga cgaggatctt ggagcactcc ttggaaactc
     3121 gacagaccct gcggtgttta ccgacctcgc gtcagtagat aactccgaat ttcagcagct
     3181 tttgaaccag ggtatcccgg tcgcgccaca tacaacggag cccatgttga tggaataccc
     3241 cgaagcaatc acgagacttg tgacgggagc gcagcggcct cccgatcccg cacccgcacc
     3301 tttgggggca cctggcctcc ctaacggact tttgagcggc gacgaggatt tctcctccat
     3361 cgccgatatg gatttctcag ccttgctgtc acagatttcc agcggctctg gcagcggcag
     3421 ccgggattcc agggaaggga tgtttttgcc gaagcctgag gccggctccg ctattagtga
     3481 cgtgtttgag ggccgcgagg tgtgccagcc aaaacgaatc cggccatttc atcctccagg
     3541 aagtccatgg gccaaccgcc cactccccgc cagcctcgca ccaacaccaa ccggtccagt
     3601 acatgagcca gtcgggtcac tgaccccggc accagtccct cagccactgg atccagcgcc
     3661 cgcagtgact cccgaggcca gtcacctgtt ggaggatccc gatgaagaga cgagccaggc
     3721 tgtcaaagcc cttcgggaga tggccgatac tgtgattccc cagaaggaag aggctgcaat
     3781 ctgtggccaa atggaccttt cccatccgcc cccaaggggc catctggatg agctgacaac
     3841 cacacttgag tccatgaccg aggatctgaa cctggactca cccctgaccc cggaattgaa
     3901 cgagattctg gataccttcc tgaacgacga gtgcctcttg catgccatgc atatcagcac
     3961 aggactgtcc atcttcgaca catctctgtt ttgaatttaa ataattcgcc cctctccctc
     4021 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     4081 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     4141 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     4201 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     4261 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     4321 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     4381 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     4441 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
     4501 gtgtgtttag tcgaggttaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     4561 ctttgaaaaa cacgatgata atatggccac aaccggatcc ggagctacca tggatagatc
     4621 cggaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     4681 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     4741 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     4801 ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg
     4861 ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     4921 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc
     4981 gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     5041 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     5101 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     5161 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     5221 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     5281 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     5341 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg
     5401 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     5461 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     5521 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg
     5581 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     5641 ataggtttaa accatgcaag cttgatatca agcttatcga taatcaacct ctggattaca
     5701 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     5761 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     5821 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     5881 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     5941 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     6001 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     6061 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     6121 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     6181 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     6241 gtcggatctc cctttgggcc gcctccccgc atcctgcagg taattctacc gggtagggga
     6301 ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg
     6361 ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc
     6421 tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc
     6481 ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag
     6541 gctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg
     6601 ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg
     6661 gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc
     6721 ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc
     6781 gggcctttcg acctgcagcc caagcttgtc gacgccacca tgaagagaga tcatcatcat
     6841 catcatcatc aagataagaa gactatgatg atgaatgaag aagacgacgg taacggcatg
     6901 gatgagcttc tagctgttct tggttacaag gttaggtcat ccgaaatggc tgatgttgct
     6961 cagaaactcg agcagcttga agttatgatg tctaatgttc aagaagacga tctttctcaa
     7021 ctcgctactg agactgttca ctataatccg gcggagcttt acacgtggct tgattctatg
     7081 ctcaccgacc ttaatcatat gtcaagcgct gcaggtgcta ccatgagcga gctgattaag
     7141 gagaacatgc acatgaagct gtacatggag ggcaccgtgg acaaccatca cttcaagtgc
     7201 acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggtggtc
     7261 gagggcggcc ctctcccctt cgccttcgac atcctggcta ctagcttcct ctacggcagc
     7321 aagaccttca tcaaccacac ccagggcatc cccgacttct tcaagcagtc cttccctgag
     7381 ggcttcacat gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag
     7441 gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc
     7501 acatccaacg gccctgtgat gcagaagaaa acactcggct gggaggcctt caccgagacg
     7561 ctgtaccccg ctgacggcgg cctggaaggc agaaacgaca tggccctgaa gctcgtgggc
     7621 gggagccatc tgatcgcaaa catcaagacc acatatagat ccaagaaacc cgctaagaac
     7681 ctcaagatgc ctggcgtcta ctatgtggac tacagactgg aaagaatcaa ggaggccaac
     7741 aacgagacct acgtcgagca gcacgaggtg gcagtggcca gatactgcga cctccctagc
     7801 aaactggggc acaagcttaa tggtggaggc tctggaatgg ccccaaagaa gaagcggaag
     7861 gtcggtatcc acggagtccc agcagccaag cggaactaca tcctgggcct ggccatcggc
     7921 atcaccagcg tgggctacgg catcatcgac tacgagacac gggacgtgat cgatgccggc
     7981 gtgcggctgt tcaaagaggc caacgtggaa aacaacgagg gcaggcggag caagagaggc
     8041 gccagaaggc tgaagcggcg gaggcggcat agaatccaga gagtgaagaa gctgctgttc
     8101 gactacaacc tgctgaccga ccacagcgag ctgagcggca tcaaccccta cgaggccaga
     8161 gtgaagggcc tgagccagaa gctgagcgag gaagagttct ctgccgccct gctgcacctg
     8221 gccaagagaa gaggcgtgca caacgtgaac gaggtggaag aggacaccgg caacgagctg
     8281 tccaccaaag agcagatcag ccggaacagc aaggccctgg aagagaaata cgtggccgaa
     8341 ctgcagctgg aacggctgaa gaaagacggc gaagtgcggg gcagcatcaa cagattcaag
     8401 accagcgact acgtgaaaga agccaaacag ctgctgaagg tgcagaaggc ctaccaccag
     8461 ctggaccaga gcttcatcga cacctacatc gacctgctgg aaacccggcg gacctactat
     8521 gagggacctg gcgagggcag ccccttcggc tggaaggaca tcaaagaatg gtacgagatg
     8581 ctgatgggcc actgcaccta cttccccgag gaactgcgga gcgtgaagta cgcctacaac
     8641 gccgacctgt acaacgccct gaacgacctg aacaatctcg tgatcaccag ggacgagaac
     8701 gagaagctgg aatattacga gaagttccag atcatcgaga acgtgttcaa gcagaagaag
     8761 aagcccaccc tgaagcagat cgccaaagaa atcctcgtga acgaagagga tattaagggc
     8821 tacagagtga ccagcaccgg caagcccgag ttcaccaacc tgaaggtgta ccacgacatc
     8881 aaggacatta ccgcccggaa agagattatt gagaacgccg agctgctgga tcagattgcc
     8941 aagatcctga ccatctacca gagcagcgag gacatccagg aagaactgac caatctgaac
     9001 tccgagctga cccaggaaga gatcgagcag atctctaatc tgaagggcta taccggcacc
     9061 cacaacctga gcctgaaggc catcaacctg atcctggacg agctgtggca caccaacgac
     9121 aaccagatcg ctatcttcaa ccggctgaag ctggtgccca agaaggtgga cctgtcccag
     9181 cagaaagaga tccccaccac cctggtggac gacttcatcc tgagccccgt cgtgaagaga
     9241 agcttcatcc agagcatcaa agtgatcaac gccatcatca agaagtacgg cctgcccaac
     9301 gacatcatta tcgagctggc ccgcgagaag aactccaagg acgcccagaa aatgatcaac
     9361 gagatgcaga agcggaaccg gcagaccaac gagcggatcg aggaaatcat ccggaccacc
     9421 ggcaaagaga acgccaagta cctgatcgag aagatcaagc tgcacgacat gcaggaaggc
     9481 aagtgcctgt acagcctgga agccatccct ctggaagatc tgctgaacaa ccccttcaac
     9541 tatgaggtgg accacatcat ccccagaagc gtgtccttcg acaacagctt caacaacaag
     9601 gtgctcgtga agcaggaaga agccagcaag aagggcaacc ggaccccatt ccagtacctg
     9661 agcagcagcg acagcaagat cagctacgaa accttcaaga agcacatcct gaatctggcc
     9721 aagggcaagg gcagaatcag caagaccaag aaagagtatc tgctggaaga acgggacatc
     9781 aacaggttct ccgtgcagaa agacttcatc aaccggaacc tggtggatac cagatacgcc
     9841 accagaggcc tgatgaacct gctgcggagc tacttcagag tgaacaacct ggacgtgaaa
     9901 gtgaagtcca tcaatggcgg cttcaccagc tttctgcggc ggaagtggaa gtttaagaaa
     9961 gagcggaaca aggggtacaa gcaccacgcc gaggacgccc tgatcattgc caacgccgat
    10021 ttcatcttca aagagtggaa gaaactggac aaggccaaaa aagtgatgga aaaccagatg
    10081 ttcgaggaaa agcaggccga gagcatgccc gagatcgaaa ccgagcagga gtacaaagag
    10141 atcttcatca ccccccacca gatcaagcac attaaggact tcaaggacta caagtacagc
    10201 caccgggtgg acaagaagcc taatagagag ctgattaacg acaccctgta ctccacccgg
    10261 aaggacgaca agggcaacac cctgatcgtg aacaatctga acggcctgta cgacaaggac
    10321 aatgacaagc tgaaaaagct gatcaacaag agccccgaaa agctgctgat gtaccaccac
    10381 gacccccaga cctaccagaa actgaagctg attatggaac agtacggcga cgagaagaat
    10441 cccctgtaca agtactacga ggaaaccggg aactacctga ccaagtactc caaaaaggac
    10501 aacggccccg tgatcaagaa gattaagtat tacggcaaca aactgaacgc ccatctggac
    10561 atcaccgacg actaccccaa cagcagaaac aaggtcgtga agctgtccct gaagccctac
    10621 agattcgacg tgtacctgga caatggcgtg tacaagttcg tgaccgtgaa gaatctggat
    10681 gtgatcaaaa aagaaaacta ctacgaagtg aatagcaagt gctatgagga agctaagaag
    10741 ctgaagaaga tcagcaacca ggccgagttt atcgcctcct tctacaacaa cgatctgatc
    10801 aagatcaacg gcgagctgta tagagtgatc ggcgtgaaca acgacctgct gaaccggatc
    10861 gaagtgaaca tgatcgacat cacctaccgc gagtacctgg aaaacatgaa cgacaagagg
    10921 ccccccagga tcattaagac aatcgcctcc aagacccaga gcattaagaa gtacagcaca
    10981 gacattctgg gcaacctgta tgaagtgaaa tctaagaagc accctcagat catcaaaaag
    11041 ggcaaaaggc cggcggccac gaaaaaggcc ggccaggcaa aaaagaaaaa gtaactcgag
    11101 ctagttaaaa gttttgttac tttatagaag aaattttgag tttttgtttt tttttaataa
    11161 ataaataaac ataaataaat tgtttgttga atttattatt agtatgtaag tgtaaatata
    11221 ataaaactta atatctattc aaattaataa ataaacctcg atatacagac cgataaaaca
    11281 catgcgtcaa ttttacgcat gattatcttt aacgtacgtc acaatatgat tatctttcta
    11341 gggttaatct agtatacgcg tatgcggccg cttaattaat ccagcttttg ttccctttag
    11401 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
    11461 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
    11521 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
    11581 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
    11641 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
    11701 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
    11761 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
    11821 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
    11881 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
    11941 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
    12001 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
    12061 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
    12121 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
    12181 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
    12241 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
    12301 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
    12361 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
    12421 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
    12481 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
    12541 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
    12601 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
    12661 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
    12721 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
    12781 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
    12841 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
    12901 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
    12961 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
    13021 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
    13081 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
    13141 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
    13201 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
    13261 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
    13321 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
    13381 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
    13441 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
    13501 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
    13561 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
    13621 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
    13681 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
    13741 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
    13801 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
    13861 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
    13921 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
    13981 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
    14041 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
    14101 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
    14161 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac
    14221 tatagggcga attggggcgc gccattctag attaacccta gaaagatagt ctgcgtaaaa
    14281 ttgacgcatg cattcttgaa atattgctct ctctttctaa atagcgcgaa tccgtcgctg
    14341 tgcatttagg acatctcagt cgccgcttgg agctcccgtg aggcgtgctt gtcaatgcgg
    14401 taagtgtcac tgattttgaa ctataacgac cgcgtgagtc aaaatgacgc atgattatct
    14461 tttacgtgac ttttaagatt taactcatac gataattata ttgttatttc atgttctact
    14521 tacgtgataa cttattatat atatattttc ttgttataga tatcaactag aatgctagca
    14581 tgggcccaga tct
//
