LOCUS       sequence-445704-       11875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LPUtopia matching RMCE donor plasmid with MONARCH 2.0 circuit with 
            crRNA targeting EGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11875)
  AUTHORS   Wan Y, Helenek C, Coraci D, Balazsi G
  TITLE     Optimizing a CRISPR-Cas13d Gene Circuit for Tunable Target RNA 
            Downregulation with Minimal Collateral RNA Cutting.
  JOURNAL   ACS Synth Biol. 2024 Oct 18;13(10):3212-3230. doi: 
            10.1021/acssynbio.4c00271. Epub 2024 Oct 8.
  PUBMED    39377757
REFERENCE   2  (bases 1 to 11875)
  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: 
            "10.1021/acssynbio.4c00271"; journalName: "ACS Synth Biol"; date: 
            "2024-10-18- 18"; volume: "13"; issue: "10"; pages: "3212-3230"
COMMENT     Sequence Label: pUt-MONARCH 2.0_crEGFP
FEATURES             Location/Qualifiers
     source          1..11875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      14..602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     503..522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(609..642)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     enhancer        753..1132
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1133..1336
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1287..1307
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    1332..1350
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1358..1376
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     intron          1559..2131
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     regulatory      2143..2152
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2800..2856
                     /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             2857..3564
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3002..3020)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3270..3289
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3574..3630
                     /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             3634..6534
                     /codon_start=1
                     /product="RNA-targeting Type VI CRISPR protein from 
                     Ruminococcus flavefaciens strain XPD3002 (Konermann et al.,
                     2018)"
                     /label=RfxCas13d
                     /translation="MIEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDS
                     IRSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLE
                     KRYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFSTV
                     YTYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYII
                     NYGNECYDILALLSGLRHWVVHNNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITN
                     ELTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRK
                     DMSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFV
                     INLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPA
                     GRDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAF
                     FKDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDE
                     NGNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQ
                     KKQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGRE
                     NAEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLE
                     EKGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKK
                     AEEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFRNKAVHLEVARYVH
                     AYINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCV
                     PFGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS"
     CDS             6544..6564
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6574..6600
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6607..6672
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_RNA        6850..6885
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     misc_RNA        6909..6944
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     primer_bind     complement(7034..7054)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(7407..7424)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7413..7637
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    7772..8166
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     rep_origin      8602..8736
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(8606..8903)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     8901..8921
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     protein_bind    complement(8949..8982)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     promoter        9049..9153
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9154..9222,9223..10014)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTASQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(9372..9391)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        10047..10546
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(10082..10105)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     10479..10498
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     5'UTR           10588..10627
                     /label=HSV TK 5'-UTR
                     /note="5' untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     CDS             10645..11775
                     /codon_start=1
                     /product="herpes simplex virus thymidine kinase"
                     /label=HSV TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MASYPCHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRLEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWQVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     primer_bind     complement(11773..11792)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    11817..11865
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 tcataggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
       61 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      121 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
      181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      601 aaaacgccga agttcctata ctttctagag aataggaact tcacgggcca gatatacgcg
      661 acgggccaga tatacgcgac gggccagata tacgcgacgg gccagatata cgcgacgggc
      721 cagatatacg cgacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
      781 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      841 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      901 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      961 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     1021 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     1081 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     1141 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     1201 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     1261 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     1321 ctatataagc atccctatca gtgatagaga tcagatctcc ctatcagtga tagagagctg
     1381 ttttgacctc catagaagac accgggaccg atccagcctc cggactctag cgtttaaact
     1441 taagcttggt acccggggat cctctagggc ctctgagcta ttccagaagt agtgaagagg
     1501 cttttttgga ggcctaggct tttgcaaaaa gctccggatc gatcctgaga acttcagggt
     1561 gagtttgggg acccttgatt gttctttctt tttcgctatt gtaaaattca tgttatatgg
     1621 agggggcaaa gttttcaggg tgttgtttag aatgggaaga tgtcccttgt atcaccatgg
     1681 accctcatga taattttgtt tctttcactt tctactctgt tgacaaccat tgtctcctct
     1741 tattttcttt tcattttctg taactttttc gttaaacttt agcttgcatt tgtaacgaat
     1801 ttttaaattc acttttgttt atttgtcaga ttgtaagtac tttctctaat cacttttttt
     1861 tcaaggcaat cagggtatat tatattgtac ttcagcacag ttttagagaa caattgttat
     1921 aattaaatga taaggtagaa tatttctgca tataaattct ggctggcgtg gaaatattct
     1981 tattggtaga aacaactaca tcctggtcat catcctgcct ttctctttat ggttacaatg
     2041 atatacactg tttgagatga ggataaaata ctctgagtcc aaaccgggcc cctctgctaa
     2101 ccatgttcat gccttcttct ttttcctaca ggtcctgcag gcgccaccat gggttctaga
     2161 ctggacaaga gcaaagtcat aaactctgct ctggaattac tcaatgaagt cggtatcgaa
     2221 ggcctgacga caaggaaact cgctcaaaag ctgggagttg agcagcctac cctgtactgg
     2281 cacgtgaaga acaagcgggc cctgctcgat gccctggcaa tcgagatgct ggacaggcat
     2341 catacccact tctgccccct ggaaggcgag tcatggcaag actttctgcg gaacaacgcc
     2401 aagtcattcc gctgtgctct cctctcacat cgcgacgggg ctaaagtgca tctcggcacc
     2461 cgcccaacag agaaacagta cgaaaccctg gaaaatcagc tcgcgttcct gtgtcagcaa
     2521 ggcttctccc tggagaacgc actgtacgct ctgtccgccg tgggccactt tacactgggc
     2581 tgcgtattgg aggatcagga gcatcaagta gcaaaagagg aaagagagac acctaccacc
     2641 gattctatgc ccccacttct gagacaagca attgagctgt tcgaccatca gggagccgaa
     2701 cctgccttcc ttttcggcct ggaactaatc atatgtggcc tggagaaaca gctaaagtgc
     2761 gaaagcggcg ggccaaaaaa gaagagaaag ggaagcggag ctactaactt cagcctgctg
     2821 aagcaggctg gagacgtgga ggagaaccct ggacctatgg tgagcaaggg cgaggaggat
     2881 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     2941 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     3001 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
     3061 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
     3121 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     3181 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
     3241 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     3301 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     3361 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
     3421 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
     3481 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
     3541 ggcggcatgg acgagctgta caagggcagc ggagctacta acttcagcct gctgaagcag
     3601 gctggagacg tggaggagaa ccctggaccc gggatgatcg agaaaaagaa gtccttcgct
     3661 aagggcatgg gcgtgaagtc cacactcgtg tccggctcca aagtgtacat gacaaccttc
     3721 gccgaaggca gcgacgccag gctggaaaag atcgtggagg gcgacagcat caggagcgtg
     3781 aatgagggcg aggccttcag cgctgaaatg gccgataaaa acgccggcta taagatcggc
     3841 aacgccaaat tcagccatcc taagggctac gccgtggtgg ctaacaaccc tctgtataca
     3901 ggacccgtcc agcaggatat gctcggcctg aaggaaactc tggaaaagag gtacttcggc
     3961 gagagcgctg atggcaatga caatatttgt atccaggtga tccataacat cctggacatt
     4021 gaaaaaatcc tcgccgaata cattaccaac gccgcctacg ccgtcaacaa tatctccggc
     4081 ctggataagg acattattgg attcggcaag ttctccacag tgtataccta cgacgaattc
     4141 aaagaccccg agcaccatag ggccgctttc aacaataacg ataagctcat caacgccatc
     4201 aaggcccagt atgacgagtt cgacaacttc ctcgataacc ccagactcgg ctatttcggc
     4261 caggcctttt tcagcaagga gggcagaaat tacatcatca attacggcaa cgaatgctat
     4321 gacattctgg ccctcctgag cggactgaga cactgggtgg tccacaacaa cgaagaagag
     4381 tccaggatct ccaggacctg gctctacaac ctcgataaga acctcgacaa cgaatacatc
     4441 tccaccctca actacctcta cgacaggatc accaatgagc tgaccaactc cttctccaag
     4501 aactccgccg ccaacgtgaa ctatattgcc gaaactctgg gaatcaaccc tgccgaattc
     4561 gccgaacaat atttcagatt cagcattatg aaagagcaga aaaacctcgg attcaatatc
     4621 accaagctca gggaagtgat gctggacagg aaggatatgt ccgagatcag gaaaaatcat
     4681 aaggtgttcg actccatcag gaccaaggtc tacaccatga tggactttgt gatttatagg
     4741 tattacatcg aagaggatgc caaggtggct gccgccaata agtccctccc cgataatgag
     4801 aagtccctga gcgagaagga tatctttgtg attaacctga ggggctcctt caacgacgac
     4861 cagaaggatg ccctctacta cgatgaagct aatagaattt ggagaaagct cgaaaatatc
     4921 atgcacaaca tcaaggaatt taggggaaac aagacaagag agtataagaa gaaggacgcc
     4981 cctagactgc ccagaatcct gcccgctggc cgtgatgttt ccgccttcag caaactcatg
     5041 tatgccctga caatgttcct ggatggcaag gagatcaacg acctcctgac caccctgatt
     5101 aataaattcg ataacatcca gagcttcctg aaggtgatgc ctctcatcgg agtcaacgct
     5161 aagttcgtgg aggaatacgc ctttttcaaa gactccgcca agatcgccga tgagctgagg
     5221 ctgatcaagt ccttcgctag aatgggagaa cctattgccg atgccaggag ggccatgtat
     5281 atcgacgcca tccgtatttt aggaaccaac ctgtcctatg atgagctcaa ggccctcgcc
     5341 gacacctttt ccctggacga gaacggaaac aagctcaaga aaggcaagca cggcatgaga
     5401 aatttcatta ttaataacgt gatcagcaat aaaaggttcc actacctgat cagatacggt
     5461 gatcctgccc acctccatga gatcgccaaa aacgaggccg tggtgaagtt cgtgctcggc
     5521 aggatcgctg acatccagaa aaaacagggc cagaacggca agaaccagat cgacaggtac
     5581 tacgaaactt gtatcggaaa ggataagggc aagagcgtga gcgaaaaggt ggacgctctc
     5641 acaaagatca tcaccggaat gaactacgac caattcgaca agaaaaggag cgtcattgag
     5701 gacaccggca gggaaaacgc cgagagggag aagtttaaaa agatcatcag cctgtacctc
     5761 accgtgatct accacatcct caagaatatt gtcaatatca acgccaggta cgtcatcgga
     5821 ttccattgcg tcgagcgtga tgctcaactg tacaaggaga aaggctacga catcaatctc
     5881 aagaaactgg aagagaaggg attcagctcc gtcaccaagc tctgcgctgg cattgatgaa
     5941 actgcccccg ataagagaaa ggacgtggaa aaggagatgg ctgaaagagc caaggagagc
     6001 attgacagcc tcgagagcgc caaccccaag ctgtatgcca attacatcaa atacagcgac
     6061 gagaagaaag ccgaggagtt caccaggcag attaacaggg agaaggccaa aaccgccctg
     6121 aacgcctacc tgaggaacac caagtggaat gtgatcatca gggaggacct cctgagaatt
     6181 gacaacaaga catgtaccct gttcaggaac aaggccgtcc atctggaagt ggccaggtat
     6241 gtccacgcct atatcaacga cattgccgag gtcaattcct acttccaact gtaccattac
     6301 atcatgcaga gaattatcat gaatgagagg tacgagaaaa gcagcggaaa ggtgtccgag
     6361 tacttcgacg ctgtgaatga cgagaagaag tacaacgata ggctcctgaa actgctgtgt
     6421 gtgcctttcg gctactgtat ccccaggttt aagaacctga gcatcgaggc cctgttcgat
     6481 aggaacgagg ccgccaagtt cgacaaggag aaaaagaaag tgtccggcaa ttccactagt
     6541 gctcccaaga aaaagcgcaa ggtaggtgga agcccagcag ctaaaagagt taaattggat
     6601 ggatccgatt ataaggatca cgatggagat tacaaggacc acgacataga ctacaaagat
     6661 gatgacgaca agtaacgtac gtaagaattc gattcgtcag tagggttgta aaggtttttc
     6721 ttttcctgag aaaacaacct tttgttttct caggttttgc tttttggcct ttccctagct
     6781 ttaaaaaaaa aaaagcaaaa ctcaccgagg cagttccata ggatggcaag gccgcgactc
     6841 tagagtcgac aagtaaaccc ctaccaactg gtcggggttt gaaaccatga tatagacgtt
     6901 gtggctgtca agtaaacccc taccaactgg tcggggtttg aaactcgacc tgcaggcatg
     6961 caagcttgat atcaggccgc aatcaacctc tggattacaa aatttgtgaa agattgactg
     7021 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     7081 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     7141 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     7201 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     7261 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     7321 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcc tcgagtctag
     7381 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     7441 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     7501 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     7561 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     7621 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     7681 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gctaaagcaa
     7741 gtaaaacctc tacaaatgtg gtatggaatt ctaaaataca gcatagcaaa actttaacct
     7801 ccaaatcaag cctctacttg aatccttttc tgagggatga ataaggcata ggcatcaggg
     7861 gctgttgcca atgtgcatta gctgtttgca gcctcacctt ctttcatgga gtttaagata
     7921 tagtgtattt tcccaaggtt tgaactagct cttcatttct ttatgtttta aatgcactga
     7981 cctcccacat tcccttttta gtaaaatatt cagaaataat ttaaatacat cattgcaatg
     8041 aaaataaatg ttttttatta ggcagaatcc agatgctcaa ggcccttcat aatatccccc
     8101 agtttagtag ttggacttag ggaacaaagg aacctttaat agaaattgga cagcaagaaa
     8161 gcgagcttct agctttagtt cctggtgtac ttgaggggga tgagttcctc aatggtggtt
     8221 ttgaccagct tgccattcat ctcaatgagc acaaagcagt caggagcata gtcagagatg
     8281 agctctctgc acatgccaca ggggctgacc accctgatgg atctgtccac ctcatcagag
     8341 taggggtgcc tgacagccac aatggtgtca aagtccttct gcccgttgct cacagcagac
     8401 ccaatggcaa tggcttcagc acagacagtg accctgccaa tgtaggcctc aatgtggaca
     8461 gcagagatga tctccccagt cttggtcctg atggccgccc cgacatggtg cttgttgtcc
     8521 tcatagagca tggtgatctt ctcagtggcg acctccacca gctccagatc ctgctgagag
     8581 atgttgaagg tcttcatggt agcctccaaa aaagcctcct cactacttct ggaatagctc
     8641 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     8701 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     8761 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     8821 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     8881 ggagcctggg gactttccac accctaactg acacacattc cacagctgcg aagagaagag
     8941 gtcaggccga agttcctata ctatttgaag aataggaact tcaagcttgc cccactgtgg
     9001 ggtggagggg acagataaaa gtaccctggc acttttcggg gaaatgtgcg cggaacccct
     9061 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     9121 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     9181 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     9241 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     9301 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     9361 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     9421 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     9481 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     9541 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     9601 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     9661 gccataccaa acgacgagcg tgacaccacg atgcccgtag caatggcaac aacgttgcgc
     9721 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     9781 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     9841 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     9901 gatggtaagc cctcccgtat cgtagttatc tacacgacgg cgagtcaggc aactatggat
     9961 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
    10021 gaccaagttt attctacggt tctaccgggt aggggaggcg cttttcccaa ggcagtctgg
    10081 agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg
    10141 cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc
    10201 gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg
    10261 caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc
    10321 tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc
    10381 tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag
    10441 gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg
    10501 cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct gcagcgaccc
    10561 gcttaacagc gtcaacagcg tgccgcagat cttggtggcg tgaaactccc gcacctcttc
    10621 ggcaagcgcc ttgtagaagc gcgtatggct tcgtacccct gccatcaaca cgcgtctgcg
    10681 ttcgaccagg ctgcgcgttc tcgcggccat agcaaccgac gtacggcgtt gcgccctcgc
    10741 cggcagcaag aagccacgga agtccgcctg gagcagaaaa tgcccacgct actgcgggtt
    10801 tatatagacg gtcctcacgg gatggggaaa accaccacca cgcaactgct ggtggccctg
    10861 ggttcgcgcg acgatatcgt ctacgtaccc gagccgatga cttactggca ggtgctgggg
    10921 gcttccgaga caatcgcgaa catctacacc acacaacacc gcctcgacca gggtgagata
    10981 tcggccgggg acgcggcggt ggtaatgaca agcgcccaga taacaatggg catgccttat
    11041 gccgtgaccg acgccgttct ggctcctcat atcggggggg aggctgggag ctcacatgcc
    11101 ccgcccccgg ccctcaccct catcttcgac cgccatccca tcgccgccct cctgtgctac
    11161 ccggccgcgc gataccttat gggcagcatg accccccagg ccgtgctggc gttcgtggcc
    11221 ctcatcccgc cgaccttgcc cggcacaaac atcgtgttgg gggcccttcc ggaggacaga
    11281 cacatcgacc gcctggccaa acgccagcgc cccggcgagc ggcttgacct ggctatgctg
    11341 gccgcgattc gccgcgttta cgggctgctt gccaatacgg tgcggtatct gcagggcggc
    11401 gggtcgtggc gggaggattg gggacagctt tcggggacgg ccgtgccgcc ccagggtgcc
    11461 gagccccaga gcaacgcggg cccacgaccc catatcgggg acacgttatt taccctgttt
    11521 cgggcccccg agttgctggc ccccaacggc gacctgtaca acgtgtttgc ctgggccttg
    11581 gacgtcttgg ccaaacgcct ccgtcccatg cacgtcttta tcctggatta cgaccaatcg
    11641 cccgccggct gccgggacgc cctgctgcaa cttacctccg ggatggtcca gacccacgtc
    11701 accacccccg gctccatacc gacgatctgc gacctggcgc gcacgtttgc ccgggagatg
    11761 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
    11821 aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcgc
//
