LOCUS       sequence-309706-       12901 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chromosomal integration of PgpdA-dSpCas9-VPR-TtrpC. Contains 1kb 
            homology arms for Aspergillus nidulans and the bar gene for 
            glufosinate selection. Filamentous fungi vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12901)
  AUTHORS   Roux I, Woodcraft C, Hu J, Wolters R, Gilchrist CLM, Chooi YH
  TITLE     CRISPR-mediated activation of biosynthetic gene clusters for 
            bioactive molecule discovery in filamentous fungi.
  JOURNAL   ACS Synth Biol. 2020 Jun 11. doi: 10.1021/acssynbio.0c00197.
  PUBMED    32526136
REFERENCE   2  (bases 1 to 12901)
  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"; journalName: "ACS Synth 
            Biol. 2020 Jun 11. doi: 10.1021/acssynbio.0c00197."
COMMENT     Sequence Label: pCRI003-pGEM-LS-Bar-PgpdA-dSpCas9-VPR-TtrpC-LS
FEATURES             Location/Qualifiers
     source          1..12901
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..552)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(575..934)
                     /label=trpC promoter
                     /note="promoter for Aspergillus nidulans trpC"
     primer_bind     complement(1673..1689)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1674..1690)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             join(1719..1745,1746..1748,1749..4232,4233..4238,4239..4304,
                     4305..4307,4308..5822)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5835..5855
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5880..6029
                     /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             6054..6074
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6504..6860
                     /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             6879..7448
                     /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"
     regulatory      8830..8839
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      8834..8843
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(8873..8892)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8874..8892)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8899..8916)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8899..8915)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8908..8930)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9056..9511
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9143..9162)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9353..9374
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9589..9693
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9694..9762,9763..10554)
                     /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(9912..9931)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10725..11313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11214..11233
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11467..11484
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11601..11622
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11637..11642,11643..11660,11661..11667)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11675..11691
                     /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)."
     primer_bind     11680..11702
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11699..11715
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11699..11715
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        11733..11751
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     11733..11750
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ctaaatctcg gtgacgggca ggaccggacg gggcggtacc ggcaggctga agtccagctg
       61 ccagaaaccc acgtcatgcc agttcccgtg cttgaagccg gccgcccgca gcatgccgcg
      121 gggggcatat ccgagcgcct cgtgcatgcg cacgctcggg tcgttgggca gcccgatgac
      181 agcgaccacg ctcttgaagc cctgtgcctc cagggacttc agcaggtggg tgtagagcgt
      241 ggagcccagt cccgtccgct ggtggcgggg ggagacgtac acggtcgact cggccgtcca
      301 gtcgtaggcg ttgcgtgcct tccaggggcc cgcgtaggcg atgccggcga cctcgccgtc
      361 cacctcggcg acgagccagg gatagcgctc ccgcagacgg acgaggtcgt ccgtccactc
      421 ctgcggttcc tgcggctcgg tacggaagtt gaccgtgctt gtctcgatgt agtggttgac
      481 gatggtgcag accgccggca tgtccgcctc ggtggcacgg cggatgtcgg ccgggcgtcg
      541 ttctgggctc atggttactt cctaatcgaa gctttgcttg ggtagaatag gtaagtcaga
      601 ttgaatctga aataaaggga ggaagggcga acttaagaag gtatgaccgg gtcgttcact
      661 taccttgctt gacaaacgca ccaagttatc gtgcaccaag cagcagatga taataatgtc
      721 ctcgttcctg tctgctaata agagtcacac ttcgagcgcc gccgctactg ctacaagtgg
      781 ggctgatctg accagttgcc taaatgaacc atcttgtcaa acgacacaaa ttttgtgctc
      841 accgcctgga cgactaaacc aaaataggca ttcattgttg acctccacta gctccagcca
      901 agcccaaaaa gtgctccttc aatatcatct tctgtcgacc atgcggagag acggacggac
      961 gcagagagaa gggctgagta ataagcgcca ctgcgccaga cagctctggc ggctctgagg
     1021 tgcagtggat gattattaat ccgggaccgg ccgcccctcc gccccgaagt ggaaaggctg
     1081 gtgtgcccct cgttgaccaa gaatctattg catcatcgga gaatatggag cttcatcgaa
     1141 tcaccggcag taagcgaagg agaatgtgaa gccaggggtg tatagccgtc ggcgaaatag
     1201 catgccatta acctaggtac agaagtccaa ttgcttccga tctggtaaaa gattcacgag
     1261 atagtacctt ctccgaagta ggtagagcga gtacccggcg cgtaagctcc ctaattggcc
     1321 catccggcat ctgtagggcg tccaaatatc gtgcctctcc tgctttgccc ggtgtatgaa
     1381 accggaaagg ccgctcagga gctggccagc ggcgcagacc gggaacacaa gctggcagtc
     1441 gacccatccg gtgctctgca ctcgacctgc tgaggtccct cagtccctgg taggcagctt
     1501 tgccccgtct gtccgcccgg tgtgtcggcg gggttgacaa ggtcgttgcg tcagtccaac
     1561 atttgttgcc atattttcct gctctcccca ccagctgctc ttttcttttc tctttctttt
     1621 cccatcttca gtatattcat cttcccatcc aagaaccttt aatcaagctt atcgataccg
     1681 tcgacctcga ctctagagga tcactcgtcg tgtcgcccat ggacaagaag tactccattg
     1741 ggctcgctat cggcacaaac agcgtcggct gggccgtcat tacggacgag tacaaggtgc
     1801 cgagcaaaaa attcaaagtt ctgggcaata ccgatcgcca cagcataaag aagaacctca
     1861 ttggcgccct cctgttcgac tccggggaga cggccgaagc cacgcggctc aaaagaacag
     1921 cacggcgcag atatacccgc agaaagaatc ggatctgcta cctgcaggag atctttagta
     1981 atgagatggc taaggtggat gactctttct tccataggct ggaggagtcc tttttggtgg
     2041 aggaggataa aaagcacgag cgccacccaa tctttggcaa tatcgtggac gaggtggcgt
     2101 accatgaaaa gtacccaacc atatatcatc tgaggaagaa gcttgtagac agtactgata
     2161 aggctgactt gcggttgatc tatctcgcgc tggcgcatat gatcaaattt cggggacact
     2221 tcctcatcga gggggacctg aacccagaca acagcgatgt cgataaactc tttatccaac
     2281 tggttcagac ttacaatcag cttttcgaag agaacccgat caacgcatcc ggagttgacg
     2341 ccaaagcaat cctgagcgct aggctgtcca aatcccggcg gctcgaaaac ctcatcgcac
     2401 agctccctgg ggagaagaag aacggcctgt ttggtaatct tatcgccctg tcactcgggc
     2461 tgacccccaa ctttaaatct aacttcgacc tggccgaaga tgccaagctt caactgagca
     2521 aagacaccta cgatgatgat ctcgacaatc tgctggccca gatcggcgac cagtacgcag
     2581 accttttttt ggcggcaaag aacctgtcag acgccattct gctgagtgat attctgcgag
     2641 tgaacacgga gatcaccaaa gctccgctga gcgctagtat gatcaagcgc tatgatgagc
     2701 accaccaaga cttgactttg ctgaaggccc ttgtcagaca gcaactgcct gagaagtaca
     2761 aggaaatttt cttcgatcag tctaaaaatg gctacgccgg atacattgac ggcggagcaa
     2821 gccaggagga attttacaaa tttattaagc ccatcttgga aaaaatggac ggcaccgagg
     2881 agctgctggt aaagcttaac agagaagatc tgttgcgcaa acagcgcact ttcgacaatg
     2941 gaagcatccc ccaccagatt cacctgggcg aactgcacgc tatcctcagg cggcaagagg
     3001 atttctaccc ctttttgaaa gataacaggg aaaagattga gaaaatcctc acatttcgga
     3061 taccctacta tgtaggcccc ctcgcccggg gaaattccag attcgcgtgg atgactcgca
     3121 aatcagaaga gaccatcact ccctggaact tcgaggaagt cgtggataag ggggcctctg
     3181 cccagtcctt catcgaaagg atgactaact ttgataaaaa tctgcctaac gaaaaggtgc
     3241 ttcctaaaca ctctctgctg tacgagtact tcacagttta taacgagctc accaaggtca
     3301 aatacgtcac agaagggatg agaaagccag cattcctgtc tggagagcag aagaaagcta
     3361 tcgtggacct cctcttcaag acgaaccgga aagttaccgt gaaacagctc aaagaagact
     3421 atttcaaaaa gattgaatgt ttcgactctg ttgaaatcag cggagtggag gatcgcttca
     3481 acgcatccct gggaacgtat cacgatctcc tgaaaatcat taaagacaag gacttcctgg
     3541 acaatgagga gaacgaggac attcttgagg acattgtcct cacccttacg ttgtttgaag
     3601 atagggagat gattgaagaa cgcttgaaaa cttacgctca tctcttcgac gacaaagtca
     3661 tgaaacagct caagaggcgc cgatatacag gatgggggcg gctgtcaaga aaactgatca
     3721 atgggatccg agacaagcag agtggaaaga caatcctgga ttttcttaag tccgatggat
     3781 ttgccaaccg gaacttcatg cagttgatcc atgatgactc tctcaccttt aaggaggaca
     3841 tccagaaagc acaagtttct ggccaggggg acagtcttca cgagcacatc gctaatcttg
     3901 caggtagccc agctatcaaa aagggaatac tgcagaccgt taaggtcgtg gatgaactcg
     3961 tcaaagtaat gggaaggcat aagcccgaga atatcgttat cgagatggcc cgagagaacc
     4021 aaactaccca gaagggacag aagaacagta gggaaaggat gaagaggatt gaagagggta
     4081 taaaagaact ggggtcccaa atccttaagg aacacccagt tgaaaacacc cagcttcaga
     4141 atgagaagct ctacctgtac tacctgcaga acggcaggga catgtacgtg gatcaggaac
     4201 tggacatcaa tcggctctcc gactacgacg tggctgctat cgtgccccag tcttttctca
     4261 aagatgattc tattgataat aaagtgttga caagatccga taaagctaga gggaagagtg
     4321 ataacgtccc ctcagaagaa gttgtcaaga aaatgaaaaa ttattggcgg cagctgctga
     4381 acgccaaact gatcacacaa cggaagttcg ataatctgac taaggctgaa cgaggtggcc
     4441 tgtctgagtt ggataaagcc ggcttcatca aaaggcagct tgttgagaca cgccagatca
     4501 ccaagcacgt ggcccaaatt ctcgattcac gcatgaacac caagtacgat gaaaatgaca
     4561 aactgattcg agaggtgaaa gttattactc tgaagtctaa gctggtctca gatttcagaa
     4621 aggactttca gttttataag gtgagagaga tcaacaatta ccaccatgcg catgatgcct
     4681 acctgaatgc agtggtaggc actgcactta tcaaaaaata tcccaagctt gaatctgaat
     4741 ttgtttacgg agactataaa gtgtacgatg ttaggaaaat gatcgcaaag tctgagcagg
     4801 aaataggcaa ggccaccgct aagtacttct tttacagcaa tattatgaat tttttcaaga
     4861 ccgagattac actggccaat ggagagattc ggaagcgacc acttatcgaa acaaacggag
     4921 aaacaggaga aatcgtgtgg gacaagggta gggatttcgc gacagtccgg aaggtcctgt
     4981 ccatgccgca ggtgaacatc gttaaaaaga ccgaagtaca gaccggaggc ttctccaagg
     5041 aaagtatcct cccgaaaagg aacagcgaca agctgatcgc acgcaaaaaa gattgggacc
     5101 ccaagaaata cggcggattc gattctccta cagtcgctta cagtgtactg gttgtggcca
     5161 aagtggagaa agggaagtct aaaaaactca aaagcgtcaa ggaactgctg ggcatcacaa
     5221 tcatggagcg atcaagcttc gaaaaaaacc ccatcgactt tctcgaggcg aaaggatata
     5281 aagaggtcaa aaaagacctc atcattaagc ttcccaagta ctctctcttt gagcttgaaa
     5341 acggccggaa acgaatgctc gctagtgcgg gcgagctgca gaaaggtaac gagctggcac
     5401 tgccctctaa atacgttaat ttcttgtatc tggccagcca ctatgaaaag ctcaaagggt
     5461 ctcccgaaga taatgagcag aagcagctgt tcgtggaaca acacaaacac taccttgatg
     5521 agatcatcga gcaaataagc gaattctcca aaagagtgat cctcgccgac gctaacctcg
     5581 ataaggtgct ttctgcttac aataagcaca gggataagcc catcagggag caggcagaaa
     5641 acattatcca cttgtttact ctgaccaact tgggcgcgcc tgcagccttc aagtacttcg
     5701 acaccaccat agacagaaag cggtacacct ctacaaagga ggtcctggac gccacactga
     5761 ttcatcagtc aattacgggg ctctatgaaa caagaatcga cctctctcag ctcggtggag
     5821 acagcagggc tgaccccaag aagaagagga aggtggaggc cagcggttcc ggacgggctg
     5881 acgcattgga cgattttgat ctggatatgc tgggaagtga cgccctcgat gattttgacc
     5941 ttgacatgct tggttcggat gcccttgatg actttgacct cgacatgctc ggcagtgacg
     6001 cccttgatga tttcgacctg gacatgctga ttaactctag aagttccgga tctccgaaaa
     6061 agaaacgcaa agttggtagc cagtacctgc ccgacaccga cgaccggcac cggatcgagg
     6121 aaaagcggaa gcggacctac gagacattca agagcatcat gaagaagtcc cccttcagcg
     6181 gccccaccga ccctagacct ccacctagaa gaatcgccgt gcccagcaga tccagcgcca
     6241 gcgtgccaaa acctgccccc cagccttacc ccttcaccag cagcctgagc accatcaact
     6301 acgacgagtt ccctaccatg gtgttcccca gcggccagat ctctcaggcc tctgctctgg
     6361 ctccagcccc tcctcaggtg ctgcctcagg ctcctgctcc tgcaccagct ccagccatgg
     6421 tgtctgcact ggctcaggca ccagcacccg tgcctgtgct ggctcctgga cctccacagg
     6481 ctgtggctcc accagcccct aaacctacac aggccggcga gggcacactg tctgaagctc
     6541 tgctgcagct gcagttcgac gacgaggatc tgggagccct gctgggaaac agcaccgatc
     6601 ctgccgtgtt caccgacctg gccagcgtgg acaacagcga gttccagcag ctgctgaacc
     6661 agggcatccc tgtggcccct cacaccaccg agcccatgct gatggaatac cccgaggcca
     6721 tcacccggct cgtgacaggc gctcagaggc ctcctgatcc agctcctgcc cctctgggag
     6781 caccaggcct gcctaatgga ctgctgtctg gcgacgagga cttcagctct atcgccgaca
     6841 tggacttctc cgcactgctg ggtagcggat cgggatctcg ggattccagg gaagggatgt
     6901 ttttgccgaa gcctgaggcc ggctccgcta ttagtgacgt gtttgagggc cgcgaggtgt
     6961 gccagccaaa acgaatccgg ccatttcatc ctccaggaag tccatgggcc aaccgcccac
     7021 tccccgccag cctcgcacca acaccaaccg gtccagtaca tgagccagtc gggtcactga
     7081 ccccggcacc agtccctcag ccactggatc cagcgcccgc agtgactccc gaggccagtc
     7141 acctgttgga ggatcccgat gaagagacga gccaggctgt caaagccctt cgggagatgg
     7201 ccgatactgt gattccccag aaggaagagg ctgcaatctg tggccaaatg gacctttccc
     7261 atccgccccc aaggggccat ctggatgagc tgacaaccac acttgagtcc atgaccgagg
     7321 atctgaacct ggactcaccc ctgaccccgg aattgaacga gattctggat accttcctga
     7381 acgacgagtg cctcttgcat gccatgcata tcagcacagg actgtccatc ttcgacacat
     7441 ctctgtttta gtgggcgacg acgacgttcg agggggggcc cggtaccgcc ccgtccggtc
     7501 ctgcccgtca ccgagatcca cttaacgtta ctgaaatcat caaacagctt gacgaatctg
     7561 gatataagat cgttggtgtc gatgtcagct ccggagttga gacaaatggt gttcaggatc
     7621 tcgataagat acgttcattt gtccaagcag caaagagtgc cttctagtga tttaatagct
     7681 ccatgtcaac aagaataaaa cgcgtttcgg gtttacctct tccagataca gctcatctgc
     7741 aatgcatgga cgtaaactcc tcttcagacc aaaggttggg tcgtcggtag gggaatatcc
     7801 ttggttgggg ccctggtcat aaatgccgta ttaccgcagt cagatgttgg gctgtcagtg
     7861 ggctagaagc agggatgctc tggtcccagt cccaggtata accagctttc ttagtgtata
     7921 tatcgaaagc agcagctatc ttacgccttg tctgggtttg gctggcttag ttcacaatgt
     7981 cctcaactca tctggtgact acgtattgtt ctcactttgg aggcggttca tcgcacgtaa
     8041 gagcctttgg atcgaaagtc aattcgcaaa tcagccaaaa taagcatcac gctaacaacc
     8101 ggccagtgaa tgctcctggg caatgacgag ctcatataag accggtagtc catttcagtc
     8161 caatgacgag tcatcgcaat acttgacccg cacatccaca tttcacaagg atgtaaggga
     8221 gccctaagat agtcaccgaa cgggctagtt gctgtgagga tcctggtgaa gttcaagtta
     8281 atgacatagg aaaatcatct ttattccagg gagaagaata ggccaatcgt ttaccctttg
     8341 ctcgttcttt ctgatggcct gtctatctac caaaaagttt gtctaggata taatctaggc
     8401 aaatgaagca agaatcaata ccatcctttc tgccctatgg tgtagactta gaaagtggat
     8461 gcactgtctg ccacagttat gccttggtca ggcgctgtga cgtaatggct cgcagccaca
     8521 attttgaatc gacagccaat tccagttcaa gccccgatac cctgtttatg tttaccaaga
     8581 caagcatcta ttatcagata ctaccaggaa attctaagag cgcgaccggc ccctgatcgg
     8641 aggttggttg tgtgtttttg ctgaactggg ggactcgcac accaaccgga cgaattctgc
     8701 acactgatac cacactcgct tcatgcaagt ttcaagccat cccttaacgc gtcagtgcat
     8761 acaaatgagt gtctatcgac ccattaacgc gctgtccaat gctgggcgac gacgacgtga
     8821 attcccgcgg ccgccatggc ggccgggagc atgcgacgtc gggcccaatt cgccctatag
     8881 tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     8941 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     9001 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg
     9061 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     9121 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     9181 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     9241 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     9301 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     9361 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     9421 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     9481 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     9541 tgtgcggtat ttcacaccgc atcaggtggc acttttcggg gaaatgtgcg cggaacccct
     9601 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     9661 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     9721 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     9781 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     9841 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     9901 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     9961 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
    10021 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
    10081 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
    10141 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
    10201 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
    10261 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
    10321 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
    10381 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
    10441 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
    10501 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
    10561 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
    10621 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
    10681 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
    10741 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
    10801 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
    10861 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
    10921 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
    10981 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
    11041 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
    11101 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
    11161 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
    11221 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
    11281 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
    11341 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
    11401 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
    11461 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
    11521 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
    11581 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
    11641 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
    11701 ggaaacagct atgaccatga ttacgccaag ctatttaggt gacactatag aatactcaag
    11761 ctatgcatcc aacgcgttgg gagctctccc atatggtcga cctgcaggcg gccgcgaatt
    11821 cctcgtcgtg tcgcccaagt gaggtaatga ttaggattgc tggaaagtta catagatata
    11881 attatatatc tctgcaatct tatgttgcaa tggtttggtg tctgcctctg ccagcgaggc
    11941 atttacgttg tccatgaccg tattaaatat aatacagcgc gcctagcatg acttttgata
    12001 ctttaaggat ctcttttaat gtcttcctgc tgttgccttt ttctggccta tgcccagaat
    12061 gcgaaaccgt cggcaaatag atgaaactag ctaagtgatt ggtcatgtca ctgttaatgt
    12121 ttagaaaatg gtattgtacc tggcatctca tcagcctgtt aaccaggtaa ggataccgat
    12181 aatatgagaa ggtattaata ggacaagcat agttagacta atgcaacgcc aaatcttggg
    12241 tatcctgtaa gagacgtttt cgggcaagca tctctcagca ctttgatcag atcccattct
    12301 cgacctgcct ggatcgcacc gataaaactg gcttgcggtt atagatgctc tagtcaaacc
    12361 atatagtcct aaatatacct gttgaaatat aacataacaa cgaaaaatgc catgacagag
    12421 cgtatctaga aaggtatata ccgtcataat acagtcaaca gtcgcggatg caggttccgc
    12481 gtactactta ctaccacagc aaaccaatgc cttcattgca taccgagcct gaaccatttt
    12541 ataacggaca ggagcagtaa cgcgccggaa caggcttggc caagagacgc aggaactcat
    12601 tgagaagccc aagttctcct agtttggtca gacagtgtcc tggcacagca cgattaattc
    12661 cccacttaca aacgatagtc tcggtatttc agacgtatat ggattagtta ttttttgttt
    12721 gattttcatg gtccagtaaa tagcgatgaa cgttgtctga gttctacaaa aagtgcagtt
    12781 cacgatcagc aacggcaaac gacgcaatgc ttcgaggtcc tgtattgggt acgtgtcatc
    12841 gtagcttact aaatgacagc aggtagatca aggtgggaca gggttattgt ctcatgagcg
    12901 g
//
