LOCUS       sequence-450828-       11852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LPUtopia matching RMCE donor plasmid with MONARCH 1.0 circuit with 
            crRNA targeting EGFP. Use BlastR for positive and HSV-TK for 
            negative selection .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11852)
  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 11852)
  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 1.0_crEGFP
FEATURES             Location/Qualifiers
     source          1..11852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          107..679
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     regulatory      691..700
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1348..1404
                     /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             1405..2112
                     /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(1550..1568)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1818..1837
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2122..2178
                     /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             2182..5082
                     /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             5092..5112
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5122..5148
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             5155..5220
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_RNA        5249..5284
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     misc_RNA        5308..5343
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     primer_bind     complement(5433..5453)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(5806..5823)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5812..6036
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    6171..6565
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     rep_origin      7001..7135
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(7005..7302)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     7300..7320
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     protein_bind    complement(7348..7381)
                     /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        7448..7552
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7553..7621,7622..8413)
                     /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(7771..7790)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        8446..8945
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8481..8504)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     8878..8897
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     5'UTR           8987..9026
                     /label=HSV TK 5'-UTR
                     /note="5' untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     CDS             9044..10174
                     /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(10172..10191)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    10216..10264
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      10288..10876
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10777..10796
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(10883..10916)
                     /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        11153..11532
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11533..11736
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11687..11707
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    11732..11750
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11758..11776
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 ccggggatcc tctagggcct ctgagctatt ccagaagtag tgaagaggct tttttggagg
       61 cctaggcttt tgcaaaaagc tccggatcga tcctgagaac ttcagggtga gtttggggac
      121 ccttgattgt tctttctttt tcgctattgt aaaattcatg ttatatggag ggggcaaagt
      181 tttcagggtg ttgtttagaa tgggaagatg tcccttgtat caccatggac cctcatgata
      241 attttgtttc tttcactttc tactctgttg acaaccattg tctcctctta ttttcttttc
      301 attttctgta actttttcgt taaactttag cttgcatttg taacgaattt ttaaattcac
      361 ttttgtttat ttgtcagatt gtaagtactt tctctaatca cttttttttc aaggcaatca
      421 gggtatatta tattgtactt cagcacagtt ttagagaaca attgttataa ttaaatgata
      481 aggtagaata tttctgcata taaattctgg ctggcgtgga aatattctta ttggtagaaa
      541 caactacatc ctggtcatca tcctgccttt ctctttatgg ttacaatgat atacactgtt
      601 tgagatgagg ataaaatact ctgagtccaa accgggcccc tctgctaacc atgttcatgc
      661 cttcttcttt ttcctacagg tcctgcaggc gccaccatgg gttctagact ggacaagagc
      721 aaagtcataa actctgctct ggaattactc aatgaagtcg gtatcgaagg cctgacgaca
      781 aggaaactcg ctcaaaagct gggagttgag cagcctaccc tgtactggca cgtgaagaac
      841 aagcgggccc tgctcgatgc cctggcaatc gagatgctgg acaggcatca tacccacttc
      901 tgccccctgg aaggcgagtc atggcaagac tttctgcgga acaacgccaa gtcattccgc
      961 tgtgctctcc tctcacatcg cgacggggct aaagtgcatc tcggcacccg cccaacagag
     1021 aaacagtacg aaaccctgga aaatcagctc gcgttcctgt gtcagcaagg cttctccctg
     1081 gagaacgcac tgtacgctct gtccgccgtg ggccacttta cactgggctg cgtattggag
     1141 gatcaggagc atcaagtagc aaaagaggaa agagagacac ctaccaccga ttctatgccc
     1201 ccacttctga gacaagcaat tgagctgttc gaccatcagg gagccgaacc tgccttcctt
     1261 ttcggcctgg aactaatcat atgtggcctg gagaaacagc taaagtgcga aagcggcggg
     1321 ccaaaaaaga agagaaaggg aagcggagct actaacttca gcctgctgaa gcaggctgga
     1381 gacgtggagg agaaccctgg acctatggtg agcaagggcg aggaggataa catggccatc
     1441 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     1501 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     1561 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     1621 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     1681 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     1741 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     1801 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     1861 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     1921 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     1981 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     2041 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     2101 gagctgtaca agggcagcgg agctactaac ttcagcctgc tgaagcaggc tggagacgtg
     2161 gaggagaacc ctggacccgg gatgatcgag aaaaagaagt ccttcgctaa gggcatgggc
     2221 gtgaagtcca cactcgtgtc cggctccaaa gtgtacatga caaccttcgc cgaaggcagc
     2281 gacgccaggc tggaaaagat cgtggagggc gacagcatca ggagcgtgaa tgagggcgag
     2341 gccttcagcg ctgaaatggc cgataaaaac gccggctata agatcggcaa cgccaaattc
     2401 agccatccta agggctacgc cgtggtggct aacaaccctc tgtatacagg acccgtccag
     2461 caggatatgc tcggcctgaa ggaaactctg gaaaagaggt acttcggcga gagcgctgat
     2521 ggcaatgaca atatttgtat ccaggtgatc cataacatcc tggacattga aaaaatcctc
     2581 gccgaataca ttaccaacgc cgcctacgcc gtcaacaata tctccggcct ggataaggac
     2641 attattggat tcggcaagtt ctccacagtg tatacctacg acgaattcaa agaccccgag
     2701 caccataggg ccgctttcaa caataacgat aagctcatca acgccatcaa ggcccagtat
     2761 gacgagttcg acaacttcct cgataacccc agactcggct atttcggcca ggcctttttc
     2821 agcaaggagg gcagaaatta catcatcaat tacggcaacg aatgctatga cattctggcc
     2881 ctcctgagcg gactgagaca ctgggtggtc cacaacaacg aagaagagtc caggatctcc
     2941 aggacctggc tctacaacct cgataagaac ctcgacaacg aatacatctc caccctcaac
     3001 tacctctacg acaggatcac caatgagctg accaactcct tctccaagaa ctccgccgcc
     3061 aacgtgaact atattgccga aactctggga atcaaccctg ccgaattcgc cgaacaatat
     3121 ttcagattca gcattatgaa agagcagaaa aacctcggat tcaatatcac caagctcagg
     3181 gaagtgatgc tggacaggaa ggatatgtcc gagatcagga aaaatcataa ggtgttcgac
     3241 tccatcagga ccaaggtcta caccatgatg gactttgtga tttataggta ttacatcgaa
     3301 gaggatgcca aggtggctgc cgccaataag tccctccccg ataatgagaa gtccctgagc
     3361 gagaaggata tctttgtgat taacctgagg ggctccttca acgacgacca gaaggatgcc
     3421 ctctactacg atgaagctaa tagaatttgg agaaagctcg aaaatatcat gcacaacatc
     3481 aaggaattta ggggaaacaa gacaagagag tataagaaga aggacgcccc tagactgccc
     3541 agaatcctgc ccgctggccg tgatgtttcc gccttcagca aactcatgta tgccctgaca
     3601 atgttcctgg atggcaagga gatcaacgac ctcctgacca ccctgattaa taaattcgat
     3661 aacatccaga gcttcctgaa ggtgatgcct ctcatcggag tcaacgctaa gttcgtggag
     3721 gaatacgcct ttttcaaaga ctccgccaag atcgccgatg agctgaggct gatcaagtcc
     3781 ttcgctagaa tgggagaacc tattgccgat gccaggaggg ccatgtatat cgacgccatc
     3841 cgtattttag gaaccaacct gtcctatgat gagctcaagg ccctcgccga caccttttcc
     3901 ctggacgaga acggaaacaa gctcaagaaa ggcaagcacg gcatgagaaa tttcattatt
     3961 aataacgtga tcagcaataa aaggttccac tacctgatca gatacggtga tcctgcccac
     4021 ctccatgaga tcgccaaaaa cgaggccgtg gtgaagttcg tgctcggcag gatcgctgac
     4081 atccagaaaa aacagggcca gaacggcaag aaccagatcg acaggtacta cgaaacttgt
     4141 atcggaaagg ataagggcaa gagcgtgagc gaaaaggtgg acgctctcac aaagatcatc
     4201 accggaatga actacgacca attcgacaag aaaaggagcg tcattgagga caccggcagg
     4261 gaaaacgccg agagggagaa gtttaaaaag atcatcagcc tgtacctcac cgtgatctac
     4321 cacatcctca agaatattgt caatatcaac gccaggtacg tcatcggatt ccattgcgtc
     4381 gagcgtgatg ctcaactgta caaggagaaa ggctacgaca tcaatctcaa gaaactggaa
     4441 gagaagggat tcagctccgt caccaagctc tgcgctggca ttgatgaaac tgcccccgat
     4501 aagagaaagg acgtggaaaa ggagatggct gaaagagcca aggagagcat tgacagcctc
     4561 gagagcgcca accccaagct gtatgccaat tacatcaaat acagcgacga gaagaaagcc
     4621 gaggagttca ccaggcagat taacagggag aaggccaaaa ccgccctgaa cgcctacctg
     4681 aggaacacca agtggaatgt gatcatcagg gaggacctcc tgagaattga caacaagaca
     4741 tgtaccctgt tcaggaacaa ggccgtccat ctggaagtgg ccaggtatgt ccacgcctat
     4801 atcaacgaca ttgccgaggt caattcctac ttccaactgt accattacat catgcagaga
     4861 attatcatga atgagaggta cgagaaaagc agcggaaagg tgtccgagta cttcgacgct
     4921 gtgaatgacg agaagaagta caacgatagg ctcctgaaac tgctgtgtgt gcctttcggc
     4981 tactgtatcc ccaggtttaa gaacctgagc atcgaggccc tgttcgatag gaacgaggcc
     5041 gccaagttcg acaaggagaa aaagaaagtg tccggcaatt ccactagtgc tcccaagaaa
     5101 aagcgcaagg taggtggaag cccagcagct aaaagagtta aattggatgg atccgattat
     5161 aaggatcacg atggagatta caaggaccac gacatagact acaaagatga tgacgacaag
     5221 taacgtacgg ccgcgactct agagtcgaca agtaaacccc taccaactgg tcggggtttg
     5281 aaaccatgat atagacgttg tggctgtcaa gtaaacccct accaactggt cggggtttga
     5341 aactcgacct gcaggcatgc aagcttgata tcaggccgca atcaacctct ggattacaaa
     5401 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     5461 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     5521 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     5581 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     5641 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     5701 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     5761 gtgttgtcct cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     5821 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     5881 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     5941 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     6001 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     6061 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     6121 tggttacgcg ctaaagcaag taaaacctct acaaatgtgg tatggaattc taaaatacag
     6181 catagcaaaa ctttaacctc caaatcaagc ctctacttga atccttttct gagggatgaa
     6241 taaggcatag gcatcagggg ctgttgccaa tgtgcattag ctgtttgcag cctcaccttc
     6301 tttcatggag tttaagatat agtgtatttt cccaaggttt gaactagctc ttcatttctt
     6361 tatgttttaa atgcactgac ctcccacatt ccctttttag taaaatattc agaaataatt
     6421 taaatacatc attgcaatga aaataaatgt tttttattag gcagaatcca gatgctcaag
     6481 gcccttcata atatccccca gtttagtagt tggacttagg gaacaaagga acctttaata
     6541 gaaattggac agcaagaaag cgagcttcta gctttagttc ctggtgtact tgagggggat
     6601 gagttcctca atggtggttt tgaccagctt gccattcatc tcaatgagca caaagcagtc
     6661 aggagcatag tcagagatga gctctctgca catgccacag gggctgacca ccctgatgga
     6721 tctgtccacc tcatcagagt aggggtgcct gacagccaca atggtgtcaa agtccttctg
     6781 cccgttgctc acagcagacc caatggcaat ggcttcagca cagacagtga ccctgccaat
     6841 gtaggcctca atgtggacag cagagatgat ctccccagtc ttggtcctga tggccgcccc
     6901 gacatggtgc ttgttgtcct catagagcat ggtgatcttc tcagtggcga cctccaccag
     6961 ctccagatcc tgctgagaga tgttgaaggt cttcatggta gcctccaaaa aagcctcctc
     7021 actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa
     7081 ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg
     7141 gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc
     7201 ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt
     7261 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc
     7321 acagctgcga agagaagagg tcaggccgaa gttcctatac tatttgaaga ataggaactt
     7381 caagcttgcc ccactgtggg gtggagggga cagataaaag taccctggca cttttcgggg
     7441 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     7501 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     7561 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     7621 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     7681 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     7741 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     7801 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     7861 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     7921 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     7981 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     8041 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcccgtagc
     8101 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     8161 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     8221 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     8281 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     8341 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     8401 taagcattgg taactgtcag accaagttta ttctacggtt ctaccgggta ggggaggcgc
     8461 ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca
     8521 caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt
     8581 tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg
     8641 ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg
     8701 tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat
     8761 agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg
     8821 ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat
     8881 tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc
     8941 tttcgacctg cagcgacccg cttaacagcg tcaacagcgt gccgcagatc ttggtggcgt
     9001 gaaactcccg cacctcttcg gcaagcgcct tgtagaagcg cgtatggctt cgtacccctg
     9061 ccatcaacac gcgtctgcgt tcgaccaggc tgcgcgttct cgcggccata gcaaccgacg
     9121 tacggcgttg cgccctcgcc ggcagcaaga agccacggaa gtccgcctgg agcagaaaat
     9181 gcccacgcta ctgcgggttt atatagacgg tcctcacggg atggggaaaa ccaccaccac
     9241 gcaactgctg gtggccctgg gttcgcgcga cgatatcgtc tacgtacccg agccgatgac
     9301 ttactggcag gtgctggggg cttccgagac aatcgcgaac atctacacca cacaacaccg
     9361 cctcgaccag ggtgagatat cggccgggga cgcggcggtg gtaatgacaa gcgcccagat
     9421 aacaatgggc atgccttatg ccgtgaccga cgccgttctg gctcctcata tcggggggga
     9481 ggctgggagc tcacatgccc cgcccccggc cctcaccctc atcttcgacc gccatcccat
     9541 cgccgccctc ctgtgctacc cggccgcgcg ataccttatg ggcagcatga ccccccaggc
     9601 cgtgctggcg ttcgtggccc tcatcccgcc gaccttgccc ggcacaaaca tcgtgttggg
     9661 ggcccttccg gaggacagac acatcgaccg cctggccaaa cgccagcgcc ccggcgagcg
     9721 gcttgacctg gctatgctgg ccgcgattcg ccgcgtttac gggctgcttg ccaatacggt
     9781 gcggtatctg cagggcggcg ggtcgtggcg ggaggattgg ggacagcttt cggggacggc
     9841 cgtgccgccc cagggtgccg agccccagag caacgcgggc ccacgacccc atatcgggga
     9901 cacgttattt accctgtttc gggcccccga gttgctggcc cccaacggcg acctgtacaa
     9961 cgtgtttgcc tgggccttgg acgtcttggc caaacgcctc cgtcccatgc acgtctttat
    10021 cctggattac gaccaatcgc ccgccggctg ccgggacgcc ctgctgcaac ttacctccgg
    10081 gatggtccag acccacgtca ccacccccgg ctccataccg acgatctgcg acctggcgcg
    10141 cacgtttgcc cgggagatgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
    10201 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt
    10261 gttcataaac gcgctcatag gatcttcttg agatcctttt tttctgcgcg taatctgctg
    10321 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
    10381 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
    10441 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
    10501 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
    10561 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
    10621 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
    10681 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
    10741 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
    10801 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
    10861 gcggagccta tggaaaaacg ccgaagttcc tatactttct agagaatagg aacttcacgg
    10921 gccagatata cgcgacgggc cagatatacg cgacgggcca gatatacgcg acgggccaga
    10981 tatacgcgac gggccagata tacgcgacgg gccagatata cgcgacgggc cagatatacg
    11041 cgacgggcca gatatacgcg acgggccaga tatacgcgac gggccagata tacgcgacgg
    11101 gccagatata cgcgacgggc cagatatacg cgacgggcca gatatacgcg ttgacattga
    11161 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
    11221 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
    11281 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
    11341 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
    11401 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
    11461 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
    11521 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
    11581 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
    11641 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
    11701 aggcgtgtac ggtgggaggt ctatataagc atccctatca gtgatagaga tcagatctcc
    11761 ctatcagtga tagagagctg ttttgacctc catagaagac accgggaccg atccagcctc
    11821 cggactctag cgtttaaact taagcttggt ac
//
