LOCUS       Exported               12585 bp ds-DNA     circular SYN 08-APR-2021
DEFINITION  SB-transposon with constitutive bi-directional promoter, one side: 
            SfiI cloning site for GOI (contains filler DNA), other side: RFP and
            blasticidin resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    pSBbi-RB
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12585)
  AUTHORS   Kowarz E, Loescher D, Marschalek R
  TITLE     Optimized Sleeping Beauty transposons rapidly generate stable 
            transgenic cell lines.
  JOURNAL   Biotechnol J. 2015 Feb 4. doi: 10.1002/biot.201400821.
  PUBMED    25650551
REFERENCE   2  (bases 1 to 12585)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jul 9, 2021 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..12585
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(272..292)
                     /label=HAP1Cas9-5'-R
                     /note="2752 57.3127747059763"
     primer_bind     319..338
                     /label=MS2-p65_HSF1-BPK-INS_R
                     /note="5469 70.3412619507065"
     polyA_signal    324..548
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     341..352
                     /label=LHA mr in54 g3 Q5 insert R
                     /note="3479 69.0316895793382"
     primer_bind     341..352
                     /label=tat pX601 52-54 lin LHA R
                     /note="3487 58.4411771725687"
     primer_bind     352..373
                     /label=polyA-2-R
                     /note="534 55.077987621628303"
     primer_bind     complement(417..438)
                     /label=HAP1Cas9-3'-F
                     /note="2750 56.5811193289025"
     primer_bind     434..452
                     /label=BGH-Rev
                     /note="1725 52.5926583040538"
     primer_bind     434..452
                     /label=ITR REV
                     /note="1934 52.6"
     primer_bind     complement(519..540)
                     /label=polyA-1-2-F
                     /note="532 57.229539140428997"
     primer_bind     complement(520..540)
                     /label=Poly A-1 -2F
                     /note="985 57.2"
     primer_bind     complement(536..554)
                     /label=BGH-Fwd
                     /note="1724 54.6433339140921"
     primer_bind     complement(536..554)
                     /label=ITR F
                     /note="1933 54.6"
     primer_bind     537..554
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     primer_bind     complement(555..572)
                     /label=DNMT3A-vector-F
                     /note="2569 57.7991327167286"
     primer_bind     complement(555..572)
                     /label=hHDAC-vector-F
                     /note="2567 57.7991327167286"
     primer_bind     complement(558..572)
                     /label=dSaCas9-VPR-linearize F
                     /note="3421 57.3847435970713"
     primer_bind     complement(558..572)
                     /label=px601 guide check F
                     /note="3220 55.2844079865314"
     primer_bind     complement(588..604)
                     /label=SBVPR-Vect-F
                     /note="3298 59.7725313917689"
     CDS             complement(608..628)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(608..628)
                     /label=Linear_BE3_vector_F
                     /note="2293 52.8356513794482"
     primer_bind     612..628
                     /label=MST Cas9 BamHI KL R
                     /note="1583 65.1764223813074"
     primer_bind     615..634
                     /label=PE2-Seq-R
                     /note="3718 54.4006949974903"
     primer_bind     complement(649..667)
                     /label=iSpyMac-IF-VECT-R
                     /note="2589 57.3645613468815"
     primer_bind     complement(650..670)
                     /label=ABE8e F
                     /note="5864"
     primer_bind     704..715
                     /label=Spp1 Sp guide4-B
                     /note="2332 54.83371905328"
     primer_bind     complement(1440..1459)
                     /label=PE2-RT_seq1_F
                     /note="5324 56.0007030937736"
     primer_bind     complement(1830..1849)
                     /label=PE2-RT_seq_F
                     /note="5323 56.3359525135072"
     primer_bind     complement(2296..2315)
                     /label=PE2-Seq-F
                     /note="3717 57.1919426367353"
     primer_bind     2705..2727
                     /label=Nme2BE-IF-VECT_R
                     /note="2702 74.323951268537"
     primer_bind     complement(2729..2741)
                     /label=hUTRN_Sa_B2-B
                     /note="751 60"
     primer_bind     complement(2780..2800)
                     /label=pcDest_Nme2_vect_F
                     /note="5288 58.8517764319731"
     primer_bind     2793..2812
                     /label=ABEmax7.10-SpRY R
                     /note="5863"
     CDS             complement(2801..6901)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the H840A mutation in the HNH catalytic domain"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(2839..2854)
                     /label=SpRY-G_PE2_VECT_F
                     /note="5377 57.2170021737229"
     primer_bind     2840..2866
                     /label=SpRY-G_PE2_INS_R
                     /note="5353 66.163333188679"
     primer_bind     complement(2873..2893)
                     /label=ng-cas9-vect-F
                     /note="2851 59.9807310429047"
     primer_bind     complement(2873..2893)
                     /label=SpVQR-VECT-F
                     /note="2823 59.9807310429047"
     primer_bind     2951..2973
                     /label=spCas9 R
                     /note="1607 56.7734156681027"
     primer_bind     3058..3080
                     /label=spCas9 R2
                     /note="1609 56.7012721400352"
     primer_bind     complement(3096..3118)
                     /label=spCas9 F
                     /note="1606 57.6423751510204"
     primer_bind     3503..3524
                     /label=SpVQR-VECT-R
                     /note="2824 59.692327761164"
     primer_bind     complement(3558..3589)
                     /label=ng-cas9-ins-F
                     /note="2853 65.2621316050305"
     primer_bind     3575..3597
                     /label=ng-cas9-vect-R
                     /note="2852 57.9817501969075"
     primer_bind     complement(3605..3627)
                     /label=spCas9 F2
                     /note="1608 54.9113985906414"
     primer_bind     3608..3635
                     /label=iSpyMac-IF-VECT-F
                     /note="2588 57.6815593641015"
     primer_bind     3627..3638
                     /label=Exon22-2B
                     /note="2901 61.8218470137284"
     primer_bind     4018..4029
                     /label=Ifi44 qPCR R
                     /note="4399 59.6827047962486"
     primer_bind     4164..4183
                     /label=SpCas9-Seq-R
                     /note="2724 55.5775200797602"
     primer_bind     4335..4346
                     /label=SaCas9-N580A_SDM_R
                     /note="5381 53.0046719275964"
     primer_bind     complement(4370..4397)
                     /label=SpRY-SpG_H840A-SDM_F
                     /note="5354 65.5129815099054"
     primer_bind     4398..4428
                     /label=SpRY-SpG_H840A-SDM1_R
                     /note="5356 66.4400092096019"
     primer_bind     4398..4415
                     /label=SpRY-SpG_H840A-SDM_R
                     /note="5355 57.1038744705631"
     primer_bind     4826..4845
                     /label=SpCas9-Seq2-R
                     /note="2725 54.6791540491694"
     primer_bind     5298..5317
                     /label=SpCas9-Seq3-R
                     /note="2726 54.4403998361641"
     primer_bind     5765..5784
                     /label=SpCas9-Seq4-R
                     /note="2727 54.8967907792937"
     primer_bind     complement(5860..5872)
                     /label=Plectin 1f_qPCR_F
                     /note="4751 60.5"
     primer_bind     6283..6293
                     /label=Linear_mCherry_ex79_rv
                     /note="1695 61.08108117472"
     primer_bind     complement(6327..6346)
                     /label=SpCas9-Seq5-R
                     /note="2728 57.2725330789406"
     primer_bind     6495..6517
                     /label=MS2 seq R
                     /note="1589 57.68545878"
     primer_bind     complement(6746..6772)
                     /label=SpRY-G_PE2_INS_F
                     /note="5352 63.9694357781862"
     primer_bind     6758..6781
                     /label=SpRY-G_PE2_VECT_R
                     /note="5351 58.026530879358"
     primer_bind     complement(6869..6883)
                     /label=SaCas9 D10 insert F
                     /note="3346 69.8546836502307"
     primer_bind     complement(6884..6902)
                     /label=NotI_intro-F
                     /note="572 69.866289147423402"
     primer_bind     complement(6884..6902)
                     /label=NotI_Cas9 fw
                     /note="543 63.3"
     primer_bind     complement(6890..6912)
                     /label=ABEmax7.10-SpRY F
                     /note="5862"
     primer_bind     6902..6926
                     /label=pcDest_Nme2_vect_R
                     /note="5289 58.1391332690075"
     CDS             complement(6902..6922)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6902..6919)
                     /label=Linear_BE3_vector_F
                     /note="2293 52.8356513794482"
     primer_bind     complement(6907..6974)
                     /label=SB-VPR-F
                     /note="3103 72.9438513044446"
     primer_bind     6956..6973
                     /label=SBVPR-Vect-R
                     /note="3299 58.6722888740028"
     primer_bind     complement(6963..6980)
                     /label=SBVPR-Vect-R
                     /note="3299 58.6722888740028"
     primer_bind     6988..7018
                     /label=inter_MS2-BPK-Ins_R
                     /note="5465 67.6585568703953"
     promoter        complement(6990..8171)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     6990..7018
                     /label=MS2-p65_HSF1-BPK-Vect_R
                     /note="5467 58.0805225444265"
     primer_bind     complement(7022..7042)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     7745..7754
                     /label=SBVPR-Vect-F
                     /note="3298 59.7725313917689"
     primer_bind     complement(8078..8097)
                     /label=CEP-Fw
                     /note="133 56.5"
     primer_bind     complement(8136..8154)
                     /label=T385M Q5 LoxP Distal R
                     /note="4538 65.8049652106758"
     primer_bind     8154..8172
                     /label=T385M Q5 LoxP Distal F
                     /note="4537 64.6491278623306"
     primer_bind     complement(8407..8417)
                     /label=mCherry-F-roundOne
                     /note="2980 68.7355826040394"
     primer_bind     8797..8820
                     /label=GFP-SLC-F
                     /note="3601 75.2713228557626"
     primer_bind     8803..8821
                     /label=Q5_GS3_linker_fw
                     /note="1691 67.7482549595673"
     primer_bind     8803..8820
                     /label=MMEJ-RB-round1-F
                     /note="3095 67.7004780040669"
     CDS             8804..9505
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MLSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     8804..8824
                     /label=tial1-GFPtagF
                     /note="3547 71.3827059761463"
     primer_bind     8804..8822
                     /label=Linear_mcherry_fw
                     /note="1687 60.3343184274323"
     primer_bind     8808..8825
                     /label=mCherry-F-roundOne
                     /note="2980 68.7355826040394"
     primer_bind     8808..8821
                     /label=GFP-knockIn-F_1
                     /note="2915 67.9077626869633"
     primer_bind     complement(8937..8957)
                     /label=mCherry_seq_rv
                     /note="1754 58.2873251883673"
     primer_bind     complement(8937..8955)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     9205..9224
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     9331..9343
                     /label=in54-10-R
                     /note="674 60.4"
     primer_bind     9399..9422
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     9430..9449
                     /label=mCherry-Fa
                     /note="2992 55.6357288607356"
     primer_bind     complement(9484..9505)
                     /label=Linear_mCherry_ex79_rv
                     /note="1695 61.08108117472"
     primer_bind     complement(9485..9505)
                     /label=CFTR-eGFP_ins-R
                     /note="2955 68.3240419368055"
     primer_bind     complement(9485..9505)
                     /label=deltaFGFP-Ins_R
                     /note="3112 64.6754549288707"
     primer_bind     complement(9486..9506)
                     /label=EGFP-AFIII-R
                     /note="2048 62.0350012969099"
     primer_bind     complement(9487..9506)
                     /label=MS2_LSD1_R
                     /note="2344 63.2340167170752"
     primer_bind     complement(9487..9505)
                     /label=tial1-GFPtagR
                     /note="3548 71.468717275925"
     primer_bind     complement(9487..9505)
                     /label=GFP-SLC-R
                     /note="3600 71.1534604393468"
     primer_bind     complement(9487..9505)
                     /label=mCherry-R-roundOne
                     /note="2981 69.5044758715562"
     primer_bind     complement(9487..9505)
                     /label=eGFP SacII KL R
                     /note="1587 66.1212602302058"
     primer_bind     complement(9487..9505)
                     /label=EGFP_HincII-R
                     /note="359 62.7635455281522"
     primer_bind     complement(9488..9506)
                     /label=DNMT3A-insert-R
                     /note="2572 62.5611725465985"
     primer_bind     complement(9490..9506)
                     /label=hHDAC-insert-R
                     /note="2566 61.8303426612664"
     primer_bind     complement(9491..9507)
                     /label=GFP-knockIn-R_1
                     /note="2916 67.9004692424559"
     primer_bind     9506..9518
                     /label=P2A HA F
                     /note="3223 60.677228597145"
     CDS             9515..9571
                     /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             9572..9970
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAMPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(9693..9714)
                     /label=HAP1Cas9-3'-R2
                     /note="2749 55.5749473208204"
     primer_bind     complement(9942..9960)
                     /label=LINC-TIDE-R
                     /note="2914 60.5564940188604"
     primer_bind     complement(10070..10089)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    10135..10190
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     polyA_site      10135..10190
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(10185..10201)
                     /label=MMEJ-RB-round1-R
                     /note="3097 66.9950748703398"
     primer_bind     complement(10189..10208)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     10299..10310
                     /label=HAP1Cas9-5'-R
                     /note="2752 57.3127747059763"
     primer_bind     complement(10582..10599)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(10631..10654)
                     /label=pTRE-Tight Sequencing Primer
                     /note="20 57.9641995195053"
     primer_bind     complement(10717..10737)
                     /label=BPK2660-U6-R
                     /note="3075 68.8520599859421"
     primer_bind     complement(10738..10752)
                     /label=Fragment.REV (2)
                     /note="2489 72.0018483055393"
     primer_bind     complement(10745..10768)
                     /label=ILMNA-LIBVECT-R
                     /note="4475 70.9052260760041"
     primer_bind     complement(10745..10767)
                     /label=sequencing_AAV_F
                     /note="341 60.452906311500001"
     rep_origin      complement(10753..11341)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10805..10824)
                     /label=ori fw
                     /note="507 60.8"
     primer_bind     complement(10833..10852)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11512..12372)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(11631..11652)
                     /label=AmpR_No_BsaI_F
                     /note="1649 64.78606981"
     primer_bind     11657..11675
                     /label=AmpR_No_BsaI_R
                     /note="1648 64.27511556"
     primer_bind     complement(11816..11827)
                     /label=PMD M2 dupl junct R
                     /note="3501 54.7912066936087"
     primer_bind     12135..12154
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12373..12477)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggatccctat acagttgaag tcggaagttt acatacactt aagttggagt cattaaaact
       61 cgtttttcaa ctactccaca aatttcttgt taacaaacaa tagttttggc aagtcagtta
      121 ggacatctac tttgtgcatg acacaagtca tttttccaac aattgtttac agacagatta
      181 tttcacttat aattcactgt atcacaattc cagtgggtca gaagtttaca tacactaagt
      241 tcgactcctc tgcagaatgc ggcgatgttt cggtaagggg tccgctatct agtaggccca
      301 gctggttctt tccgcctcag aagccataga gcccaccgca tccccagcat gcctgctatt
      361 gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca
      421 cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc
      481 accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga
      541 aggcacagtc gaggctgatc agcgagctct agagaattga tccccaagct tggcctgaca
      601 ggccttagac tttcctcttc ttcttgggct cgaattcgct gccgtcggcg gttctttttg
      661 agccgccaga gggtgatgaa ttttctatga ggagggtaga ggtgtctgga gtctctgtga
      721 tggctgcctt tcgggccgct tggtcagcca tccggttgcc tctagcctcg gcgctgtgtc
      781 ccttttgatg tcctggacaa tggattatgc taagtctttt gggcagaaag agggctttta
      841 gtagggccaa gatctcgtct ttatttttga tctctttgcc ttctgatgtg agccacccac
      901 gccttctgta tatttctcca tggatatggg cagtagcaaa agcataacgg ctatcagtat
      961 aaacatttag cttcttacct tctgccatct ttagggcctg ggtgagtgct atcagttcag
     1021 cccgctgagc ggatgtcccg gctggcaggg ctttagccca gattacctcg gtctcggtgg
     1081 tcaccgcagc tcccgcctta cgctgtccct cttgtaagag actgcttcca tccgtgtacc
     1141 aggtgtggtc ggcgtctggg agcggctggt ccgttaggtc gggtcgggtt ccgtgggctt
     1201 cggccaggat atcaaggcag ttgtgttgca gcccttcctc aggcagtggg agcagcgtag
     1261 ccgggttcag ggctaccacc ggtccgaact ggacccggtc cgtgtccaaa agcaaggcct
     1321 gatagtgagt catccgggcg ttggaaagcc agcggtcggg gggttgtttg actagtgcct
     1381 ctactgcatg gggggccaga atgactagtg gctgtcccat ggttagcttg cctgcatcct
     1441 ttgtcagtac ggcaatggct gctaccatcc gtaggcaagg gggccaccca gctgctactg
     1501 ggtctagctt tttggacagg taggccaccg gccgacgcca aggtcccagt ttttgcgtta
     1561 ggacaccttt ggcgtagccc tgcttctcgt cgacaaagag ttcaaagggc ttagtcaaat
     1621 ctggcaaccc cagggctggg gcagttagaa gagcttgctt gatttcttga taggcctttt
     1681 gttggtctgg gccccaatta aacagagtcc ccggtttggt gagagggtac aggggggctg
     1741 ccatttctgc aaacccaggg atgaagaggc gacagaagcc tgccttccct aggaactccc
     1801 ttagttgtcg aggggtcttc ggagtaggct gccccatcac agtctctttt ctggcctcag
     1861 tcagccatct ctgaccctct tttagaagat accccagata cttgacctgt ttctggcaaa
     1921 tttgggcttt cttggccgag gcccgatacc cgaggttccc tagggtttgt aacagggccc
     1981 gagtaccttg ttggcagtct agctcagaag tggcggccag cagtaagtca tccacgtact
     2041 gtagcaggat caagtctggg tgctggatcc ggaagtctgc taggtctctg tgcagtgcct
     2101 cattaaacag ggtgggactg tttttgaaac cctgtgggag tctggtccag gtcaattgtc
     2161 ctgagattcc catctctgga tctctccact caaaggcgaa gagaggctga ctggtggggt
     2221 ggagtctcag gcagaaaaag gcatccttta aatcaagcac agtgtaccac tggtgggacg
     2281 gtgggagccc gctcaagagg ttgtaagggt tgggcacggt ggggtggatg tcttccaccc
     2341 gcttgttgac ttctctcaga tcctggacag gcctataatc attagtccct ggtttcttaa
     2401 cgggtagcag gggcgtgttc cagggggact ggcagggtac cagtattccc tggtccaaca
     2461 gtctctgtat gtggggcttg atccccagtc tggcttcttg tgacatgggg tattgtttta
     2521 tggacacggg ggtagaggtt gctttcagag gtatgatcag aggagcttgg cgaactgcca
     2581 gtcccatgcc cccggtttcc gcccaggcct gaggaaaatc agacagccat gtggacccta
     2641 gagaaacatc tggctctttt gaggtctcat gtagccgata ctcatcttct atatttaggg
     2701 tgctgctgcc gccgctgctg ccgccactgc tctctggtgt tgctgactcg cttgttcctg
     2761 gtgtctcgct gccagaggat cctccgctag atcctccaga gtcacctccc agctgagaca
     2821 ggtcgatccg tgtctcgtac aggccggtga tgctctggtg gatcagggtg gcgtccagca
     2881 cctctttggt gctggtgtac ctcttccggt cgatggtggt gtcaaagtac ttgaaggcgg
     2941 caggggctcc cagattggtc agggtaaaca ggtggatgat attctcggcc tgctctctga
     3001 tgggcttatc ccggtgcttg ttgtaggcgg acagcacttt gtccagatta gcgtcggcca
     3061 ggatcactct cttggagaac tcgctgatct gctcgatgat ctcgtccagg tagtgcttgt
     3121 gctgttccac aaacagctgt ttctgctcat tatcctcggg ggagcccttc agcttctcat
     3181 agtggctggc caggtacagg aagttcacat atttggaggg cagggccagt tcgtttccct
     3241 tctgcagttc gccggcagag gccagcattc tcttccggcc gttttccagc tcgaacaggg
     3301 agtacttagg cagcttgatg atcaggtcct ttttcacttc tttgtagccc ttggcttcca
     3361 gaaagtcgat gggattcttc tcgaagctgc ttctttccat gatggtgatc cccagcagct
     3421 ctttcacact cttcagtttc ttggacttgc ccttttccac tttggccacc accagcacag
     3481 aataggccac ggtggggctg tcgaagccgc cgtacttctt agggtcccag tccttctttc
     3541 tggcgatcag cttatcgctg ttcctcttgg gcaggataga ctctttgctg aagccgcctg
     3601 tctgcacctc ggtctttttc acgatattca cttggggcat gctcagcact ttccgcacgg
     3661 tggcaaaatc ccggccctta tcccacacga tctccccggt ttcgccgttt gtctcgatca
     3721 gaggccgctt ccggatctcg ccgttggcca gggtaatctc ggtcttgaaa aagttcatga
     3781 tgttgctgta gaagaagtac ttggcggtag ccttgccgat ttcctgctcg ctcttggcga
     3841 tcatcttccg cacgtcgtac accttgtagt cgccgtacac gaactcgctt tccagcttag
     3901 ggtacttttt gatcagggcg gttcccacga cggcgttcag gtaggcgtcg tgggcgtggt
     3961 ggtagttgtt gatctcgcgc actttgtaaa actggaaatc cttccggaaa tcggacacca
     4021 gcttggactt cagggtgatc actttcactt cccggatcag cttgtcattc tcgtcgtact
     4081 tagtgttcat ccgggagtcc aggatctgtg ccacgtgctt tgtgatctgc cgggtttcca
     4141 ccagctgtct cttgatgaag ccggccttat ccagttcgct caggccgcct ctctcggcct
     4201 tggtcagatt gtcgaacttt ctctgggtaa tcagcttggc gttcagcagc tgccgccagt
     4261 agttcttcat cttcttcacg acctcttcgg agggcacgtt gtcgctcttg ccccggttct
     4321 tgtcgcttct ggtcagcacc ttgttgtcga tggagtcgtc cttcagaaag ctctgaggca
     4381 cgatagcgtc cacatcgtag tcggacagcc ggttgatgtc cagttcctgg tccacgtaca
     4441 tatcccgccc attctgcagg tagtacaggt acagcttctc gttctgcagc tgggtgtttt
     4501 ccacggggtg ttctttcagg atctggctgc ccagctcttt gatgccctct tcgatccgct
     4561 tcattctctc gcggctgttc ttctgtccct tctgggtggt ctggttctct ctggccattt
     4621 cgatcacgat gttctcgggc ttgtgccggc ccatcacttt cacgagctcg tccaccacct
     4681 tcactgtctg caggatgccc ttcttaatgg cggggctgcc ggccagattg gcaatgtgct
     4741 cgtgcaggct atcgccctgg ccggacacct gggctttctg gatgtcctct ttaaaggtca
     4801 ggctgtcgtc gtggatcagc tgcatgaagt ttctgttggc gaagccgtcg gacttcagga
     4861 aatccaggat tgtcttgccg gactgcttgt cccggatgcc gttgatcagc ttccggctca
     4921 gcctgcccca gccggtgtat ctccgccgct tcagctgctt catcactttg tcgtcgaaca
     4981 ggtgggcata ggttttcagc cgttcctcga tcatctctct gtcctcaaac agtgtcaggg
     5041 tcagcacgat atcttccaga atgtcctcgt tttcctcatt gtccaggaag tccttgtcct
     5101 tgataatttt cagcagatcg tggtatgtgc ccagggaggc gttgaaccga tcttccacgc
     5161 cggagatttc cacggagtcg aagcactcga ttttcttgaa gtagtcctct ttcagctgct
     5221 tcacggtcac tttccggttg gtcttgaaca gcaggtccac gatggccttt ttctgctcgc
     5281 cgctcaggaa ggcgggcttt ctcattccct cggtcacgta tttcactttg gtcagctcgt
     5341 tatacacggt gaagtactcg tacagcaggc tgtgcttggg cagcaccttc tcgttgggca
     5401 ggttcttatc gaagttggtc atccgctcga tgaagctctg ggcggaagcg cccttgtcca
     5461 ccacttcctc gaagttccag ggggtgatgg tttcctcgct ctttctggtc atccaggcga
     5521 atctgctgtt tcccctggcc agagggccca cgtagtaggg gatgcggaag gtcaggatct
     5581 tctcgatctt ttcccggttg tccttcagga atgggtaaaa atcttcctgc cgccgcagaa
     5641 tggcgtgcag ctctcccagg tggatctggt gggggatgct gccgttgtcg aaggtccgct
     5701 gcttccgcag caggtcctct ctgttcagct tcacgagcag ttcctcggtg ccgtccatct
     5761 tttccaggat gggcttgatg aacttgtaga actcttcctg gctggctccg ccgtcaatgt
     5821 agccggcgta gccgttcttg ctctggtcga agaaaatctc tttgtacttc tcaggcagct
     5881 gctgccgcac gagagctttc agcagggtca ggtcctggtg gtgctcgtcg tatctcttga
     5941 tcatagaggc gctcaggggg gccttggtga tctcggtgtt cactctcagg atgtcgctca
     6001 gcaggatggc gtcggacagg ttcttggcgg ccagaaacag gtcggcgtac tggtcgccga
     6061 tctgggccag caggttgtcc aggtcgtcgt cgtaggtgtc cttgctcagc tgcagtttgg
     6121 catcctcggc caggtcgaag ttgctcttga agttgggggt caggcccagg ctcagggcaa
     6181 tcaggtttcc gaacaggcca ttcttcttct cgccgggcag ctgggcgatc agattttcca
     6241 gccgtctgct cttgctcagt ctggcagaca ggatggcctt ggcgtccacg ccgctggcgt
     6301 tgatggggtt ttcctcgaac agctggttgt aggtctgcac cagctggatg aacagcttgt
     6361 ccacgtcgct gttgtcgggg ttcaggtcgc cctcgatcag gaagtggccc cggaacttga
     6421 tcatgtgggc cagggccaga tagatcagcc gcaggtcggc cttgtcggtg ctgtccacca
     6481 gtttctttct caggtggtag atggtggggt acttctcgtg gtaggccacc tcgtccacga
     6541 tgttgccgaa gatggggtgc cgctcgtgct tcttatcctc ttccaccagg aaggactctt
     6601 ccagtctgtg gaagaagctg tcgtccacct tggccatctc gttgctgaag atctcttgca
     6661 gatagcagat ccggttcttc cgtctggtgt atcttcttct ggcggttctc ttcagccggg
     6721 tggcctcggc tgtttcgccg ctgtcgaaca gcagggctcc gatcaggttc ttcttgatgc
     6781 tgtgccggtc ggtgttgccc agcaccttga atttcttgct gggcaccttg tactcgtcgg
     6841 tgatcacggc ccagcccaca gagttggtgc cgatgtccag gccgatgctg tacttcttgt
     6901 cgactttccg cttcttcttt ggtgactcga actcgcttcc gtcggctgtc cgtttcatgg
     6961 tggcctcaga ggccagcttg gggtagtttt cacgacacct taaatggaag aaaaaaactt
     7021 tgaaccactg tctgaggctt gagaatgaac caagatccaa actcaaaaag ggcaaattcc
     7081 aaggagaatt acatcaagtg ccaagctggc ctaacttcag tctccaccca ctcagtatgg
     7141 ggaaactcca tcgcataaaa cccctccccc caacctaaag acgacgtact ccaaaagctc
     7201 gagaactaat cgaggtgcct ggacggcgcc cggtactccg tggagtcaca tgaagcgacg
     7261 gctgaggacg gaaaggccct tttcctttgt gtgggtgact cacccgcccg ctctcccgag
     7321 cgccgcgtcc tccattttga gctccctgca gcagggccgg gaagcggcca tctttccgct
     7381 cacgcaactg gtgccgaccg ggccagcctt gccgcccagg gcggggcgat acacggcggc
     7441 gcgaggccag gcaccagagc aggccggcca gcttgagact acccccgtcc gattctcggt
     7501 ggccgcgctc gcaggccccg cctcgccgaa catgtgcgct gggacgcacg ggccccgtcg
     7561 ccgcccgcgg ccccaaaaac cgaaatacca gtgtgcagat cttggcccgc atttacaaga
     7621 ctatcttgcc agaaaaaaag cgtcgcagca ggtcatcaaa aattttaaat ggctagagac
     7681 ttatcgaaag cagcgagaca ggcgcgaagg tgccaccaga ttcgcacgcg gcggccccag
     7741 cgcccaggcc aagcctcaac tcaagcacga ggcgaagggg ctccttaagc gcaaggcctc
     7801 gaactctccc acccacttcc aacccgaagc tcgggatcaa gaatcacgta ctgcagccag
     7861 gggcgtggaa gtaattcaag gcacgcaagg gccataaccc gtaaagaggc caggcccgcg
     7921 ggaaccacac acggcactta cctgtgttct ggcggcaaac ccgttgcgaa aaagaacgtt
     7981 cacggcgact actgcactta tatacggttc tcccccaccc tcgggaaaaa ggcggagcca
     8041 gtacacgaca tcactttccc agtttacccc gcgccacctt ctctaggcac cggttcaatt
     8101 gccgacccct ccccccaact tctcggggac tgtgggcgat gtgcgctctg cccactgacg
     8161 ggcaccggag cctctaggcg agaccctgtc tcacaaaata aagtaagccc ggactgagtg
     8221 cggaaaggcg ggcctggcgg gtctggtctc cccatgcggg ccaccagagg ccctgcagcc
     8281 ttcagtcgct tgaaggggta atggcgcttc cactcacaaa catggcggac agagcgtgtg
     8341 aacgagatga acagcccctc aaaaatatgg ccgccgaggc tggacggccg tgccccagca
     8401 gcaccgcctc cgcgccccac gtgatctctc gccgggcaca gcgctgaccg cggaggtcca
     8461 accggaagaa tgtccggatt ggacattcgg aagagggccc gccttccctg gggaatctct
     8521 gcgcacgcgc agaacgcttc gaccaatgaa aacacaggaa gccgtccgcg caaccgcgtt
     8581 gcgtcacttc tgccgcccct gtttcaaggt atatagccgt agacggaact tcgcctttct
     8641 ctcggcctta gcgccatttt tttgggtgag tgttttttgg ttcctgcgtt gggattccgt
     8701 gtacaatcca tagacatctg acctcggcac ttagcatcat cacagcaaac taactgtagc
     8761 ctttctctct ttccctgtag aaacctctgc acctgaggcc accatgttga gcaagggcga
     8821 ggaggtcatc aaagagttca tgcgcttcaa ggtgcgcatg gagggctcca tgaacggcca
     8881 tgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     8941 gctgaaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt
     9001 catgtacggc tccaaggcgt acgtgaagca ccccgccgac atccccgatt acaagaagct
     9061 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggtctggt
     9121 gaccgtgacc caggactcct ccctgcagga cggcacgctg atctacaagg tgaagatgcg
     9181 cggcaccaac ttcccccccg acggccccgt aatgcagaag aagacaatgg gctgggaggc
     9241 ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag ggcgagatcc accaggccct
     9301 gaagctgaag gacggcggcc actacctggt ggagttcaag accatctaca tggccaagaa
     9361 gcccgtgcaa ctgcccggct actactacgt ggacaccaag ctggacatca cctcccacaa
     9421 cgaggactac accatcgtgg aacagtacga gcgctccgag ggccgccacc acctgttcct
     9481 gtacggcatg gacgagctgt acaagggcag tggagctact aacttcagcc tgctgaagca
     9541 ggctggtgac gtcgaggaga atcctggccc catggccatg cctttgtctc aagaagaatc
     9601 caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag
     9661 cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca
     9721 ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc
     9781 tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg
     9841 cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga
     9901 cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg
     9961 ggagggctaa ttcgaaggcc tgtcgtgaat tcactcctca ggtgcaggct gcctatcaga
    10021 aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct
    10081 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
    10141 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
    10201 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
    10261 catatgccat atgctggctg ccatgaacta gctactcggg accccttacc gaaacatcgc
    10321 cgcattctgc agaggagtcg agtgtatgta aacttctgac ccactgggaa tgtgatgaaa
    10381 gaaataaaag ctgaaatgaa tcattctctc tactattatt ctgatatttc acattcttaa
    10441 aataaagtgg tgatcctaac tgacctaaga cagggaattt ttactaggat taaatgtcag
    10501 gaattgtgaa aaagtgagtt taaatgtatt tggctaaggt gtatgtaaac ttccgacttc
    10561 aactgtatag ggatccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
    10621 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
    10681 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
    10741 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
    10801 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
    10861 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
    10921 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
    10981 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
    11041 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
    11101 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
    11161 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
    11221 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
    11281 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
    11341 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
    11401 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
    11461 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
    11521 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
    11581 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
    11641 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
    11701 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
    11761 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
    11821 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
    11881 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
    11941 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
    12001 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
    12061 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
    12121 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
    12181 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
    12241 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
    12301 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
    12361 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
    12421 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
    12481 atttccccga aaagtgccac ctgatgcggt gtgaaatacc gcacagatgc gtaaggagaa
    12541 aataccgcat caggaaattg taagcgttaa tattttgtta aaatt
//
