LOCUS       sequence-445954-       11573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LPUtopia matching RMCE donor plasmid with mCherry-P2A-RfxCas13d 
            Negative-Feedback circuit. Use BlastR for positive selection and 
            HSV-TK for negative selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11573)
  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 11573)
  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-mNF-Cas13d
FEATURES             Location/Qualifiers
     source          1..11573
                     /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"
     primer_bind     complement(5334..5354)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(5707..5724)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5713..5937
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    6072..6466
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     rep_origin      6902..7036
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(6906..7203)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     7201..7221
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     protein_bind    complement(7249..7282)
                     /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        7349..7453
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7454..7522,7523..8314)
                     /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(7672..7691)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        8347..8846
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8382..8405)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     8779..8798
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     5'UTR           8888..8927
                     /label=HSV TK 5'-UTR
                     /note="5' untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     CDS             8945..10075
                     /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(10073..10092)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    10117..10165
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      10189..10777
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10678..10697
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(10784..10817)
                     /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        10874..11253
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11254..11457
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11408..11428
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    11453..11471
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11479..11497
                     /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 agagtcgacc tgcaggcatg caagcttgat atcaggccgc
     5281 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     5341 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     5401 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     5461 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     5521 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     5581 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     5641 ttgggcactg acaattccgt ggtgttgtcc tcgagtctag agggcccgtt taaacccgct
     5701 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     5761 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     5821 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     5881 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     5941 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     6001 cattaagcgc ggcgggtgtg gtggttacgc gctaaagcaa gtaaaacctc tacaaatgtg
     6061 gtatggaatt ctaaaataca gcatagcaaa actttaacct ccaaatcaag cctctacttg
     6121 aatccttttc tgagggatga ataaggcata ggcatcaggg gctgttgcca atgtgcatta
     6181 gctgtttgca gcctcacctt ctttcatgga gtttaagata tagtgtattt tcccaaggtt
     6241 tgaactagct cttcatttct ttatgtttta aatgcactga cctcccacat tcccttttta
     6301 gtaaaatatt cagaaataat ttaaatacat cattgcaatg aaaataaatg ttttttatta
     6361 ggcagaatcc agatgctcaa ggcccttcat aatatccccc agtttagtag ttggacttag
     6421 ggaacaaagg aacctttaat agaaattgga cagcaagaaa gcgagcttct agctttagtt
     6481 cctggtgtac ttgaggggga tgagttcctc aatggtggtt ttgaccagct tgccattcat
     6541 ctcaatgagc acaaagcagt caggagcata gtcagagatg agctctctgc acatgccaca
     6601 ggggctgacc accctgatgg atctgtccac ctcatcagag taggggtgcc tgacagccac
     6661 aatggtgtca aagtccttct gcccgttgct cacagcagac ccaatggcaa tggcttcagc
     6721 acagacagtg accctgccaa tgtaggcctc aatgtggaca gcagagatga tctccccagt
     6781 cttggtcctg atggccgccc cgacatggtg cttgttgtcc tcatagagca tggtgatctt
     6841 ctcagtggcg acctccacca gctccagatc ctgctgagag atgttgaagg tcttcatggt
     6901 agcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     6961 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     7021 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
     7081 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
     7141 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     7201 accctaactg acacacattc cacagctgcg aagagaagag gtcaggccga agttcctata
     7261 ctatttgaag aataggaact tcaagcttgc cccactgtgg ggtggagggg acagataaaa
     7321 gtaccctggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     7381 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     7441 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     7501 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     7561 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     7621 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     7681 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     7741 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     7801 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     7861 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     7921 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     7981 tgacaccacg atgcccgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     8041 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     8101 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     8161 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     8221 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     8281 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt attctacggt
     8341 tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc
     8401 cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg
     8461 gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc
     8521 tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag
     8581 tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag
     8641 gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg
     8701 gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa
     8761 ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct
     8821 cctcttcctc atctccgggc ctttcgacct gcagcgaccc gcttaacagc gtcaacagcg
     8881 tgccgcagat cttggtggcg tgaaactccc gcacctcttc ggcaagcgcc ttgtagaagc
     8941 gcgtatggct tcgtacccct gccatcaaca cgcgtctgcg ttcgaccagg ctgcgcgttc
     9001 tcgcggccat agcaaccgac gtacggcgtt gcgccctcgc cggcagcaag aagccacgga
     9061 agtccgcctg gagcagaaaa tgcccacgct actgcgggtt tatatagacg gtcctcacgg
     9121 gatggggaaa accaccacca cgcaactgct ggtggccctg ggttcgcgcg acgatatcgt
     9181 ctacgtcccc gagccgatga cttactggca ggtgctgggg gcttccgaga caatcgcgaa
     9241 catctacacc acacaacacc gcctcgacca gggtgagata tcggccgggg acgcggcggt
     9301 ggtaatgaca agcgcccaga taacaatggg catgccttat gccgtgaccg acgccgttct
     9361 ggctcctcat atcggggggg aggctgggag ctcacatgcc ccgcccccgg ccctcaccct
     9421 catcttcgac cgccatccca tcgccgccct cctgtgctac ccggccgcgc gataccttat
     9481 gggcagcatg accccccagg ccgtgctggc gttcgtggcc ctcatcccgc cgaccttgcc
     9541 cggcacaaac atcgtgttgg gggcccttcc ggaggacaga cacatcgacc gcctggccaa
     9601 acgccagcgc cccggcgagc ggcttgacct ggctatgctg gccgcgattc gccgcgttta
     9661 cgggctgctt gccaatacgg tgcggtatct gcagggcggc gggtcgtggc gggaggattg
     9721 gggacagctt tcggggacgg ccgtgccgcc ccagggtgcc gagccccaga gcaacgcggg
     9781 cccacgaccc catatcgggg acacgttatt taccctgttt cgggcccccg agttgctggc
     9841 ccccaacggc gacctgtaca acgtgtttgc ctgggccttg gacgtcttgg ccaaacgcct
     9901 ccgtcccatg cacgtcttta tcctggatta cgaccaatcg cccgccggct gccgggacgc
     9961 cctgctgcaa cttacctccg ggatggtcca gacccacgtc accacccccg gctccatacc
    10021 gacgatctgc gacctggcgc gcacgtttgc ccgggagatg ggggaggcta actgaaacac
    10081 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
    10141 cgcacgggtg ttgggtcgtt tgttcataaa cgcgctcata ggatcttctt gagatccttt
    10201 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
    10261 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
    10321 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
    10381 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
    10441 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
    10501 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
    10561 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
    10621 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
    10681 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
    10741 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccgaagttc ctatactttc
    10801 tagagaatag gaacttcacg ggccagatat acgcgacggg ccagatatac gcgacgggcc
    10861 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
    10921 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
    10981 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
    11041 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
    11101 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
    11161 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
    11221 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
    11281 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
    11341 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
    11401 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag catccctatc
    11461 agtgatagag atcagatctc cctatcagtg atagagagct gttttgacct ccatagaaga
    11521 caccgggacc gatccagcct ccggactcta gcgtttaaac ttaagcttgg tac
//
