LOCUS       Exported               12506 bp ds-DNA     circular SYN 08-APR-2021
DEFINITION  SB-transposon with inducible SfiI cloning site for GOI (contains 
            firefly luciferase) and constitutive expression of RFP, rtTA and 
            blasticidin resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    pSBtet-RB
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12506)
  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 12506)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jul 9, 2021 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..12506
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        295..607
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    303..321
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    339..357
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    374..392
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    410..428
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    446..464
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    481..499
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    517..535
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     580..604
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     663..737
                     /label=SB-VPR-F
                     /note="3103 72.9438513044446"
     regulatory      680..685
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     complement(718..742)
                     /label=pcDest_Nme2_vect_R
                     /note="5289 58.1391332690075"
     CDS             722..742
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     725..742
                     /label=Linear_BE3_vector_F
                     /note="2293 52.8356513794482"
     primer_bind     742..760
                     /label=NotI_intro-F
                     /note="572 69.866289147423402"
     primer_bind     742..760
                     /label=NotI_Cas9 fw
                     /note="543 63.3"
     CDS             743..4843
                     /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     761..775
                     /label=SaCas9 D10 insert F
                     /note="3346 69.8546836502307"
     primer_bind     complement(863..886)
                     /label=SpRY-G_PE2_VECT_R
                     /note="5351 58.026530879358"
     primer_bind     872..898
                     /label=SpRY-G_PE2_INS_F
                     /note="5352 63.9694357781862"
     primer_bind     complement(1127..1149)
                     /label=MS2 seq R
                     /note="1589 57.68545878"
     primer_bind     1298..1317
                     /label=SpCas9-Seq5-R
                     /note="2728 57.2725330789406"
     primer_bind     complement(1860..1879)
                     /label=SpCas9-Seq4-R
                     /note="2727 54.8967907792937"
     primer_bind     complement(2327..2346)
                     /label=SpCas9-Seq3-R
                     /note="2726 54.4403998361641"
     primer_bind     complement(2799..2818)
                     /label=SpCas9-Seq2-R
                     /note="2725 54.6791540491694"
     primer_bind     complement(3216..3246)
                     /label=SpRY-SpG_H840A-SDM1_R
                     /note="5356 66.4400092096019"
     primer_bind     complement(3229..3246)
                     /label=SpRY-SpG_H840A-SDM_R
                     /note="5355 57.1038744705631"
     primer_bind     3247..3274
                     /label=SpRY-SpG_H840A-SDM_F
                     /note="5354 65.5129815099054"
     primer_bind     complement(3461..3480)
                     /label=SpCas9-Seq-R
                     /note="2724 55.5775200797602"
     primer_bind     complement(4009..4036)
                     /label=iSpyMac-IF-VECT-F
                     /note="2588 57.6815593641015"
     primer_bind     4017..4039
                     /label=spCas9 F2
                     /note="1608 54.9113985906414"
     primer_bind     complement(4047..4069)
                     /label=ng-cas9-vect-R
                     /note="2852 57.9817501969075"
     primer_bind     4055..4086
                     /label=ng-cas9-ins-F
                     /note="2853 65.2621316050305"
     primer_bind     complement(4120..4141)
                     /label=SpVQR-VECT-R
                     /note="2824 59.692327761164"
     primer_bind     4526..4548
                     /label=spCas9 F
                     /note="1606 57.6423751510204"
     primer_bind     complement(4564..4586)
                     /label=spCas9 R2
                     /note="1609 56.7012721400352"
     primer_bind     complement(4671..4693)
                     /label=spCas9 R
                     /note="1607 56.7734156681027"
     primer_bind     4751..4771
                     /label=ng-cas9-vect-F
                     /note="2851 59.9807310429047"
     primer_bind     4751..4771
                     /label=SpVQR-VECT-F
                     /note="2823 59.9807310429047"
     primer_bind     complement(4778..4804)
                     /label=SpRY-G_PE2_INS_R
                     /note="5353 66.163333188679"
     primer_bind     4790..4805
                     /label=SpRY-G_PE2_VECT_F
                     /note="5377 57.2170021737229"
     primer_bind     complement(4832..4851)
                     /label=ABEmax7.10-SpRY R
                     /note="5863"
     primer_bind     4844..4864
                     /label=pcDest_Nme2_vect_F
                     /note="5288 58.8517764319731"
     primer_bind     complement(4917..4939)
                     /label=Nme2BE-IF-VECT_R
                     /note="2702 74.323951268537"
     primer_bind     5329..5348
                     /label=PE2-Seq-F
                     /note="3717 57.1919426367353"
     primer_bind     5795..5814
                     /label=PE2-RT_seq_F
                     /note="5323 56.3359525135072"
     primer_bind     6185..6204
                     /label=PE2-RT_seq1_F
                     /note="5324 56.0007030937736"
     primer_bind     6974..6994
                     /label=ABE8e F
                     /note="5864"
     primer_bind     6977..6995
                     /label=iSpyMac-IF-VECT-R
                     /note="2589 57.3645613468815"
     primer_bind     complement(7010..7029)
                     /label=PE2-Seq-R
                     /note="3718 54.4006949974903"
     CDS             7016..7036
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     7016..7036
                     /label=Linear_BE3_vector_F
                     /note="2293 52.8356513794482"
     primer_bind     complement(7016..7032)
                     /label=MST Cas9 BamHI KL R
                     /note="1583 65.1764223813074"
     polyA_signal    7075..7196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7086..7105)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7140..7159
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             7919..8620
                     /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     complement(8052..8070)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     8320..8339
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     8514..8537
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             8630..8686
                     /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             8687..9430
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             9437..9493
                     /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             9494..9892
                     /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(9947..9964)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9953..10177
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(10503..10520)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10674..11262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10754..10773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11433..12293)
                     /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     12056..12075
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12294..12398)
                     /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 agacgagttt
      301 actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg
      361 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt
      421 gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgagttta
      481 tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta
      541 tgtcgattag gcgtgtacgg tgggcgccta taaaagcaga gctcgtttag tgaaccgtca
      601 gatcgcctgg agccaattcc aactcttttg ccttatacct actttccgta ccacttccta
      661 ccctcgaaag gcctctgagg ccaccatgaa acggacagcc gacggaagcg agttcgagtc
      721 accaaagaag aagcggaaag tcgacaagaa gtacagcatc ggcctggaca tcggcaccaa
      781 ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt
      841 gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga
      901 cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag
      961 acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga
     1021 cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga
     1081 gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac
     1141 catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat
     1201 ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct
     1261 gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca
     1321 gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc
     1381 cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa
     1441 gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag
     1501 caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga
     1561 cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa
     1621 gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa
     1681 ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct
     1741 gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca
     1801 gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa
     1861 gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa
     1921 cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat
     1981 ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa
     2041 ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc
     2101 tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac
     2161 cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg
     2221 gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct
     2281 gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat
     2341 gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa
     2401 gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg
     2461 cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata
     2521 ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga
     2581 cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga
     2641 acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg
     2701 gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca
     2761 gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat
     2821 gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc
     2881 cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa
     2941 gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca
     3001 caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca
     3061 gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca
     3121 gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta
     3181 ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc
     3241 cgactacgat gtggacgcta tcgtgcctca gagctttctg aaggacgact ccatcgacaa
     3301 caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga
     3361 ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca
     3421 gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc
     3481 cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat
     3541 cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa
     3601 agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa
     3661 agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg
     3721 aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa
     3781 ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc
     3841 caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa
     3901 cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg
     3961 ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat
     4021 cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag
     4081 gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt
     4141 cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc
     4201 caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt
     4261 cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct
     4321 gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct
     4381 ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa
     4441 cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca
     4501 gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag
     4561 cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta
     4621 caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac
     4681 cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa
     4741 gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg
     4801 cctgtacgag acacggatcg acctgtctca gctgggaggt gactctggag gatctagcgg
     4861 aggatcctct ggcagcgaga caccaggaac aagcgagtca gcaacaccag agagcagtgg
     4921 cggcagcagc ggcggcagca gcaccctaaa tatagaagat gagtatcggc tacatgagac
     4981 ctcaaaagag ccagatgttt ctctagggtc cacatggctg tctgattttc ctcaggcctg
     5041 ggcggaaacc gggggcatgg gactggcagt tcgccaagct cctctgatca tacctctgaa
     5101 agcaacctct acccccgtgt ccataaaaca ataccccatg tcacaagaag ccagactggg
     5161 gatcaagccc cacatacaga gactgttgga ccagggaata ctggtaccct gccagtcccc
     5221 ctggaacacg cccctgctac ccgttaagaa accagggact aatgattata ggcctgtcca
     5281 ggatctgaga gaagtcaaca agcgggtgga agacatccac cccaccgtgc ccaaccctta
     5341 caacctcttg agcgggctcc caccgtccca ccagtggtac actgtgcttg atttaaagga
     5401 tgcctttttc tgcctgagac tccaccccac cagtcagcct ctcttcgcct ttgagtggag
     5461 agatccagag atgggaatct caggacaatt gacctggacc agactcccac agggtttcaa
     5521 aaacagtccc accctgttta atgaggcact gcacagagac ctagcagact tccggatcca
     5581 gcacccagac ttgatcctgc tacagtacgt ggatgactta ctgctggccg ccacttctga
     5641 gctagactgc caacaaggta ctcgggccct gttacaaacc ctagggaacc tcgggtatcg
     5701 ggcctcggcc aagaaagccc aaatttgcca gaaacaggtc aagtatctgg ggtatcttct
     5761 aaaagagggt cagagatggc tgactgaggc cagaaaagag actgtgatgg ggcagcctac
     5821 tccgaagacc cctcgacaac taagggagtt cctagggaag gcaggcttct gtcgcctctt
     5881 catccctggg tttgcagaaa tggcagcccc cctgtaccct ctcaccaaac cggggactct
     5941 gtttaattgg ggcccagacc aacaaaaggc ctatcaagaa atcaagcaag ctcttctaac
     6001 tgccccagcc ctggggttgc cagatttgac taagcccttt gaactctttg tcgacgagaa
     6061 gcagggctac gccaaaggtg tcctaacgca aaaactggga ccttggcgtc ggccggtggc
     6121 ctacctgtcc aaaaagctag acccagtagc agctgggtgg cccccttgcc tacggatggt
     6181 agcagccatt gccgtactga caaaggatgc aggcaagcta accatgggac agccactagt
     6241 cattctggcc ccccatgcag tagaggcact agtcaaacaa ccccccgacc gctggctttc
     6301 caacgcccgg atgactcact atcaggcctt gcttttggac acggaccggg tccagttcgg
     6361 accggtggta gccctgaacc cggctacgct gctcccactg cctgaggaag ggctgcaaca
     6421 caactgcctt gatatcctgg ccgaagccca cggaacccga cccgacctaa cggaccagcc
     6481 gctcccagac gccgaccaca cctggtacac ggatggaagc agtctcttac aagagggaca
     6541 gcgtaaggcg ggagctgcgg tgaccaccga gaccgaggta atctgggcta aagccctgcc
     6601 agccgggaca tccgctcagc gggctgaact gatagcactc acccaggccc taaagatggc
     6661 agaaggtaag aagctaaatg tttatactga tagccgttat gcttttgcta ctgcccatat
     6721 ccatggagaa atatacagaa ggcgtgggtg gctcacatca gaaggcaaag agatcaaaaa
     6781 taaagacgag atcttggccc tactaaaagc cctctttctg cccaaaagac ttagcataat
     6841 ccattgtcca ggacatcaaa agggacacag cgccgaggct agaggcaacc ggatggctga
     6901 ccaagcggcc cgaaaggcag ccatcacaga gactccagac acctctaccc tcctcataga
     6961 aaattcatca ccctctggcg gctcaaaaag aaccgccgac ggcagcgaat tcgagcccaa
     7021 gaagaagagg aaagtctaag gcctgtcagg ccaagcttcc atcgatagac atgataagat
     7081 acattgatga gtttggacaa accacaacaa gaatgcagtg aaaaaaatgc tttatttgtg
     7141 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     7201 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa
     7261 gcaagtaaaa cctctacaaa tgtggtacct aggcgagacc ctgtctcaca aaataaagta
     7321 agcccggact gagtgcggaa aggcgggcct ggcgggtctg gtctccccat gcgggccacc
     7381 agaggccctg cagccttcag tcgcttgaag gggtaatggc gcttccactc acaaacatgg
     7441 cggacagagc gtgtgaacga gatgaacagc ccctcaaaaa tatggccgcc gaggctggac
     7501 ggccgtgccc cagcagcacc gcctccgcgc cccacgtgat ctctcgccgg gcacagcgct
     7561 gaccgcggag gtccaaccgg aagaatgtcc ggattggaca ttcggaagag ggcccgcctt
     7621 ccctggggaa tctctgcgca cgcgcagaac gcttcgacca atgaaaacac aggaagccgt
     7681 ccgcgcaacc gcgttgcgtc acttctgccg cccctgtttc aaggtatata gccgtagacg
     7741 gaacttcgcc tttctctcgg ccttagcgcc atttttttgg gtgagtgttt tttggttcct
     7801 gcgttgggat tccgtgtaca atccatagac atctgacctc ggcacttagc atcatcacag
     7861 caaactaact gtagcctttc tctctttccc tgtagaaacc tctgcacctg aggccaccat
     7921 gttgagcaag ggcgaggagg tcatcaaaga gttcatgcgc ttcaaggtgc gcatggaggg
     7981 ctccatgaac ggccatgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     8041 cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat
     8101 cctgtccccc cagttcatgt acggctccaa ggcgtacgtg aagcaccccg ccgacatccc
     8161 cgattacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     8221 ggacggcggt ctggtgaccg tgacccagga ctcctccctg caggacggca cgctgatcta
     8281 caaggtgaag atgcgcggca ccaacttccc ccccgacggc cccgtaatgc agaagaagac
     8341 aatgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga
     8401 gatccaccag gccctgaagc tgaaggacgg cggccactac ctggtggagt tcaagaccat
     8461 ctacatggcc aagaagcccg tgcaactgcc cggctactac tacgtggaca ccaagctgga
     8521 catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgct ccgagggccg
     8581 ccaccacctg ttcctgtacg gcatggacga gctgtacaag ggcagtggag ctactaactt
     8641 cagcctgctg aagcaggctg gtgacgtcga ggagaatcct ggccccatgt ctagactgga
     8701 caagagcaaa gtcataaacg gcgctctgga attactcaat ggagtcggta tcgaaggcct
     8761 gacgacaagg aaactcgctc aaaagctggg agttgagcag cctaccctgt actggcacgt
     8821 gaagaacaag cgggccctgc tcgatgccct gccaatcgag atgctggaca ggcatcatac
     8881 ccacttctgc cccctggaag gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc
     8941 attccgctgt gctctcctct cacatcgcga cggggctaaa gtgcatctcg gcacccgccc
     9001 aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt
     9061 ctccctggag aacgcactgt acgctctgtc cgccgtgggc cactttacac tgggctgcgt
     9121 attggaggaa caggagcatc aagtagcaaa agaggaaaga gagacaccta ccaccgattc
     9181 tatgccccca cttctgagac aagcaattga gctgttcgac cggcagggag ccgaacctgc
     9241 cttccttttc ggcctggaac taatcatatg tggcctggag aaacagctaa agtgcgaaag
     9301 cggcgggccg gccgacgccc ttgacgattt tgacttagac atgctcccag ccgatgccct
     9361 tgacgacttt gaccttgata tgctgcctgc tgacgctctt gacgattttg accttgacat
     9421 gctccccggg tccggcgcta ctaacttcag cctgctgaag caggctggtg acgtcgagga
     9481 gaatcctggt cccatggcca tgcctttgtc tcaagaagaa tccaccctca ttgaaagagc
     9541 aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct
     9601 ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg
     9661 tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat
     9721 cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt
     9781 gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac
     9841 ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct aattcgaagg
     9901 cctgtcgtga agcttgggga tcaattctct agagctcgct gatcagcctc gactgtgcct
     9961 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
    10021 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
    10081 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
    10141 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
    10201 tgggcctact agctactcgg gaccccttac cgaaacatcg ccgcattctg cagaggagtc
    10261 gagtgtatgt aaacttctga cccactggga atgtgatgaa agaaataaaa gctgaaatga
    10321 atcattctct ctactattat tctgatattt cacattctta aaataaagtg gtgatcctaa
    10381 ctgacctaag acagggaatt tttactagga ttaaatgtca ggaattgtga aaaagtgagt
    10441 ttaaatgtat ttggctaagg tgtatgtaaa cttccgactt caactgtata gggatccgct
    10501 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
    10561 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
    10621 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
    10681 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
    10741 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
    10801 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
    10861 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
    10921 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
    10981 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
    11041 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
    11101 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
    11161 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
    11221 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
    11281 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
    11341 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
    11401 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
    11461 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
    11521 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
    11581 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
    11641 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
    11701 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
    11761 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
    11821 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
    11881 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
    11941 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
    12001 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
    12061 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
    12121 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
    12181 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
    12241 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
    12301 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
    12361 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
    12421 cctgatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
    12481 gtaagcgtta atattttgtt aaaatt
//
