LOCUS       sequence-233281-       13953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Safe harbor site 231 knock-in vector with BlastR-dCas9-VPR 
            expression cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13953)
  AUTHORS   Pellenz S, Phelps M, Tang W, Hovde BT, Sinit RB, Fu W, Li H, Chen E,
            Monnat RJ Jr
  TITLE     New Human Chromosomal Sites with ""Safe Harbor"" Potential for 
            Targeted Transgene Insertion.
  JOURNAL   Hum Gene Ther. 2019 Jul;30(7):814-828. doi: 10.1089/hum.2018.169. 
            Epub 2019 Mar 28.
  PUBMED    30793977
REFERENCE   2  (bases 1 to 13953)
  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: 
            "10.1089/hum.2018.169"; journalName: "Hum Gene Ther"; date: 
            "2019-07"; volume: "30"; issue: "7"; pages: "814-828"
COMMENT     Sequence Label: pSH231-EF1-BLST-dCas9-VPR
FEATURES             Location/Qualifiers
     source          1..13953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(268..289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     480..499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     713..735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     727..744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   874..936
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    1974..2030
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attP site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     promoter        2065..3243
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2296..3234
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3191..3211
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(3255..3288)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3297..3306
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3303..3698
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             3705..3761
                     /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             3777..3824
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(3825..3848,3849..3851,3852..6407,6408..6410,
                     6411..7925)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7986..8006
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    8025..8049
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             8079..8228
                     /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             8253..8273
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8703..9059
                     /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
                     EDFSSIADMDFSALL"
     CDS             9078..9647
                     /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"
     polyA_signal    9720..9775
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(10457..10477)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    10523..10644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10560..10579)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10614..10633
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   11407..11441
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(11932..11948)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11932..11948)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11945..11967)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11956..11972
                     /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(11980..11986,11987..12004,12005..12010))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12025..12046
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12163..12180)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12334..12922)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12414..12433)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13093..13884,13885..13953))
                     /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     13716..13735
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggccagac tagaggtacc cattgagttc
     1381 tgaggtggtt tgtcatacag caatggcaat cagatatatc cacacaaata taattttagt
     1441 ttatatttct gctactgcag ttctcatctt attctgagga tatgtgacaa aataattctt
     1501 tcaaaaatat tgatgctgtg ctagattact attttgaatg aattactaga caaatacttc
     1561 atatgtatct tttttttttt tttttttttt tttttttttt ttttttgaga cggagtctcg
     1621 ctctgtcgcc caggctggag tgcagtggcg ggatctcggc tcactgcaat atgtatctta
     1681 ttatgtgggt ttacacatta tttatcttat tgatttaact tcaaaactaa actttagttt
     1741 agctcttggg ccctatctgg aaagggtcat cttttaatca ccattaaatc actgaagtca
     1801 tcagtttatt caaagtactc tgcacaaatt agcattcttt agtggttgtg aaataaatag
     1861 actttaaact tatcattaat attcccaatg gtactatggg ggaggcaaaa ttttctgtct
     1921 tcttagtggt tttttttttt tggctagggc taaggattaa actgtcaagc ttcgaggtgg
     1981 tttgtctggt caaccaccgc ggtctcagtg gtgtacggta caaaccccga gtcgaccttc
     2041 taggtcttga aaggagtggg aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg
     2101 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg
     2161 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     2221 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     2281 gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt
     2341 tatggccctt gcgtgccttg aattacttcc acctggctgc agtacgtgat tcttgatccc
     2401 gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg agccccttcg
     2461 cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg
     2521 caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa tttttgatga
     2581 cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca agatctgcac
     2641 actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca
     2701 tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa
     2761 gctggccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg
     2821 gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct tcccggccct
     2881 gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc
     2941 acacaaagga aaagggcctt tccgtcctca gccgtcgctt catgtgactc cacggagtac
     3001 cgggcgccgt ccaggcacct cgattagttc tcgagctttt ggagtacgtc gtctttaggt
     3061 tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga gactgaagtt
     3121 aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga gtttggatct
     3181 tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt tcaggtgtcg
     3241 tgaggaatta gcttataact tcgtatagca tacattatac gaagttatcc ctgcaggcca
     3301 ccatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa
     3361 tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg
     3421 gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg
     3481 tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg
     3541 gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc
     3601 tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga
     3661 ttcgtgaatt gctgccctct ggttatgtgt gggagggcgc tagcgccacc aacttcagcc
     3721 tgctgaagca ggccggcgac gtggaggaga accccggccc cctaggcgcc accatgaaaa
     3781 ggccggcggc cacgaaaaag gccggccagg caaaaaagaa aaaggacaag aagtacagca
     3841 tcggcctggc catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg
     3901 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc
     3961 tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa
     4021 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca
     4081 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg
     4141 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg
     4201 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg
     4261 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc
     4321 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc
     4381 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg
     4441 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg
     4501 cccagctgcc cggcgagaag aagaatggcc tgttcggcaa cctgattgcc ctgagcctgg
     4561 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga
     4621 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg
     4681 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga
     4741 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg
     4801 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt
     4861 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag
     4921 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg
     4981 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca
     5041 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg
     5101 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc
     5161 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca
     5221 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt
     5281 ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg
     5341 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag
     5401 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg
     5461 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg
     5521 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt
     5581 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc
     5641 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg
     5701 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag
     5761 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga
     5821 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg
     5881 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg
     5941 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc
     6001 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc
     6061 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga
     6121 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg
     6181 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc
     6241 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg
     6301 aactggacat caaccggctg tccgactacg atgtggacca catcgtgcct cagagctttc
     6361 tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaaggcc cggggcaaga
     6421 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc
     6481 tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg
     6541 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga
     6601 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg
     6661 acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc
     6721 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg
     6781 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg
     6841 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc
     6901 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca
     6961 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg
     7021 gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc
     7081 tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca
     7141 aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg
     7201 accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg
     7261 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca
     7321 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct
     7381 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg
     7441 aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg
     7501 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg
     7561 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg
     7621 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc
     7681 tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg
     7741 agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact
     7801 ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc
     7861 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag
     7921 gcgacagcgc tggaggaggt ggaagcggag gaggaggaag cggaggagga ggtagcggag
     7981 aattccctaa gaaaaagagg aaggtgcttg acgcgttaat atcaacaagt ttgtacaaaa
     8041 aagcaggcta caaagaggcc agcggttccg gacgggctga cgcattggac gattttgatc
     8101 tggatatgct gggaagtgac gccctcgatg attttgacct tgacatgctt ggttcggatg
     8161 cccttgatga ctttgacctc gacatgctcg gcagtgacgc ccttgatgat ttcgacctgg
     8221 acatgctgat taactctaga agttccggat ctccgaaaaa gaaacgcaaa gttggtagcc
     8281 agtacctgcc cgacaccgac gaccggcacc ggatcgagga aaagcggaag cggacctacg
     8341 agacattcaa gagcatcatg aagaagtccc ccttcagcgg ccccaccgac cctagacctc
     8401 cacctagaag aatcgccgtg cccagcagat ccagcgccag cgtgccaaaa cctgcccccc
     8461 agccttaccc cttcaccagc agcctgagca ccatcaacta cgacgagttc cctaccatgg
     8521 tgttccccag cggccagatc tctcaggcct ctgctctggc tccagcccct cctcaggtgc
     8581 tgcctcaggc tcctgctcct gcaccagctc cagccatggt gtctgcactg gctcaggcac
     8641 cagcacccgt gcctgtgctg gctcctggac ctccacaggc tgtggctcca ccagccccta
     8701 aacctacaca ggccggcgag ggcacactgt ctgaagctct gctgcagctg cagttcgacg
     8761 acgaggatct gggagccctg ctgggaaaca gcaccgatcc tgccgtgttc accgacctgg
     8821 ccagcgtgga caacagcgag ttccagcagc tgctgaacca gggcatccct gtggcccctc
     8881 acaccaccga gcccatgctg atggaatacc ccgaggccat cacccggctc gtgacaggcg
     8941 ctcagaggcc tcctgatcca gctcctgccc ctctgggagc accaggcctg cctaatggac
     9001 tgctgtctgg cgacgaggac ttcagctcta tcgccgatat ggatttctca gccttgctgg
     9061 gctctggcag cggcagccgg gattccaggg aagggatgtt tttgccgaag cctgaggccg
     9121 gctccgctat tagtgacgtg tttgagggcc gcgaggtgtg ccagccaaaa cgaatccggc
     9181 catttcatcc tccaggaagt ccatgggcca accgcccact ccccgccagc ctcgcaccaa
     9241 caccaaccgg tccagtacat gagccagtcg ggtcactgac cccggcacca gtccctcagc
     9301 cactggatcc agcgcccgca gtgactcccg aggccagtca cctgttggag gatcccgatg
     9361 aagagacgag ccaggctgtc aaagcccttc gggagatggc cgatactgtg attccccaga
     9421 aggaagaggc tgcaatctgt ggccaaatgg acctttccca tccgccccca aggggccatc
     9481 tggatgagct gacaaccaca cttgagtcca tgaccgagga tctgaacctg gactcacccc
     9541 tgaccccgga attgaacgag attctggata ccttcctgaa cgacgagtgc ctcttgcatg
     9601 ccatgcatat cagcacagga ctgtccatct tcgacacatc tctgttttga ggatcctttt
     9661 tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta
     9721 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     9781 aaggacattc tagaagtgcc ccaactgggg taacctttga gttctctcag ttgggggact
     9841 agtgcgtgga caaagagcag taaattgtga aaatggaaca aggcaagggg gcttagacta
     9901 cagtagttaa tcatcaagaa gtgacaaaaa aaataatggt tagttaataa gatttgttta
     9961 agcagatttc tcccagcttt agctctctgt ctctggtgat cagaatgcac tccttccttc
    10021 agactcagtg agcacatctt ccacatggaa gatttcttct ctagctttta ggaaatccag
    10081 agaacccttt ttgtatctgc tgtttttttt tttttaaatg tctttaactc aaaacaattt
    10141 atgtgccagg atgacatatc tttggataat gtgttctgaa ctccttcagt acatacgtat
    10201 ataaattaaa gcaaatattt tttatgataa gctggcataa tagtttcata atttaatcac
    10261 tgatttaaaa atttaattaa aattattttt tgatattttg tgtaataatt tttgaggagt
    10321 atcttttgtg cttaatgagt ggcagatgac acccatgttc ttagctacgc gttaactaag
    10381 taagcggccg ccccggtatc gacaatcaac ctctggatta caaaatttgt gaaagattga
    10441 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
    10501 tgtatcatgc gttaactaac taaacttgtt tattgcagct tataatggtt acaaataaag
    10561 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
    10621 gtccaaactc atcaatgtat cttatcatgt ctggaattga ctcaaatgat gtcaattagt
    10681 ctatcagaag ctcatctggt ctcccttccg ggggacaaga catccctgtt taatatttaa
    10741 acagcagtgt tcccaaactg ggttcttata tcccttgctc tggtcaacca ggttgcaggg
    10801 tttcctgtcc tcacaggaac gaagtcccta aagaaacagt ggcagccagg tttagccccg
    10861 gaattgactg gattcctttt ttagggccca ttggtatggc tttttccccg tatcccccca
    10921 ggtgtctgca ggctcaaaga gcagcgagaa gcgttcagag gaaagcgatc ccgtgccacc
    10981 ttccccgtgc ccgggctgtc cccgcacgct gccggctcgg ggatgcgggg ggagcgccgg
    11041 accggagcgg agccccgggc ggctcgctgc tgccccctag cgggggaggg acgtaattac
    11101 atccctgggg gctttggggg ggggctgtcc ctgatatcta taacaagaaa atatatatat
    11161 aataagttat cacgtaagta gaacatgaaa taacaatata attatcgtat gagttaaatc
    11221 ttaaaagtca cgtaaaagat aatcatgcgt cattttgact cacgcggtcg ttatagttca
    11281 aaatcagtga cacttaccgc attgacaagc acgcctcacg ggagctccaa gcggcgactg
    11341 agatgtccta aatgcacagc gacggattcg cgctatttag aaagagagag caatatttca
    11401 agaatgcatg cgtcaatttt acgcagacta tctttctagg gttaatctag ctgcatcagg
    11461 atcatatcgt cgggtctttt ttccggctca gtcatcgccc aagctggcgc tatctgggca
    11521 tcggggagga agaagcccgt gccttttccc gcgaggttga agcggcatgg aaagagtttg
    11581 ccgaggatga ctgctgctgc attgacgttg agcgaaaacg cacgtttacc atgatgattc
    11641 gggaaggtgt ggccatgcac gcctttaacg gtgaactgtt cgttcaggcc acctgggata
    11701 ccagttcgtc gcggcttttc cggacacagt tccggatggt cagcccgaag cgcatcagca
    11761 acccgaacaa taccggcgac agccggaact gccgtgccgg tgtgcagatt aatgacagcg
    11821 gtgcggcgct gggatattac gtcagcgagg acgggtatcc tggctggatg ccgcagaaat
    11881 ggacatggat accccgtgag ttacccggcg ggcgcgcttg gcgtaatcat ggtcatagct
    11941 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
    12001 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
    12061 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
    12121 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
    12181 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
    12241 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
    12301 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
    12361 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
    12421 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
    12481 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
    12541 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
    12601 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
    12661 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
    12721 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
    12781 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
    12841 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
    12901 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
    12961 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
    13021 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
    13081 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
    13141 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
    13201 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
    13261 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
    13321 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
    13381 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
    13441 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
    13501 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
    13561 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
    13621 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
    13681 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
    13741 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
    13801 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
    13861 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
    13921 aataagggcg acacggaaat gttgaatact cat
//
