LOCUS       sequence-450930-       11811 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LPUtopia matching RMCE donor plasmid with mCherry-P2A-RfxCas13d 
            Negative-Feedback circuit. Include a 3' UTR Triplex motif as a 
            template backbone for MONARCH 2.0 system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11811)
  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 11811)
  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-Triplex
FEATURES             Location/Qualifiers
     source          1..11811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        246..449
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     400..420
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    445..463
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    471..489
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     intron          672..1244
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     regulatory      1256..1265
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1913..1969
                     /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             1970..2677
                     /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(2115..2133)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2383..2402
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2687..2743
                     /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             2747..5647
                     /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             5657..5677
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5687..5713
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             5720..5785
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(6047..6067)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(6420..6437)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6426..6650
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    6785..7179
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     rep_origin      7615..7749
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(7619..7916)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     7914..7934
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     protein_bind    complement(7962..7995)
                     /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        8062..8166
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8167..8235,8236..9027)
                     /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(8385..8404)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        9060..9559
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(9095..9118)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     9492..9511
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     5'UTR           9601..9640
                     /label=HSV TK 5'-UTR
                     /note="5' untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     CDS             9658..10788
                     /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(10786..10805)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    10830..10878
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      10902..11490
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11391..11410
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(11497..11530)
                     /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        join(11677..11811,1..245)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
       61 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      121 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      181 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      241 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      301 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      361 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      421 tacggtggga ggtctatata agcatcccta tcagtgatag agatcagatc tccctatcag
      481 tgatagagag ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccggactc
      541 tagcgtttaa acttaagctt ggtacccggg gatcctctag ggcctctgag ctattccaga
      601 agtagtgaag aggctttttt ggaggcctag gcttttgcaa aaagctccgg atcgatcctg
      661 agaacttcag ggtgagtttg gggacccttg attgttcttt ctttttcgct attgtaaaat
      721 tcatgttata tggagggggc aaagttttca gggtgttgtt tagaatggga agatgtccct
      781 tgtatcacca tggaccctca tgataatttt gtttctttca ctttctactc tgttgacaac
      841 cattgtctcc tcttattttc ttttcatttt ctgtaacttt ttcgttaaac tttagcttgc
      901 atttgtaacg aatttttaaa ttcacttttg tttatttgtc agattgtaag tactttctct
      961 aatcactttt ttttcaaggc aatcagggta tattatattg tacttcagca cagttttaga
     1021 gaacaattgt tataattaaa tgataaggta gaatatttct gcatataaat tctggctggc
     1081 gtggaaatat tcttattggt agaaacaact acatcctggt catcatcctg cctttctctt
     1141 tatggttaca atgatataca ctgtttgaga tgaggataaa atactctgag tccaaaccgg
     1201 gcccctctgc taaccatgtt catgccttct tctttttcct acaggtcctg caggcgccac
     1261 catgggttct agactggaca agagcaaagt cataaactct gctctggaat tactcaatga
     1321 agtcggtatc gaaggcctga cgacaaggaa actcgctcaa aagctgggag ttgagcagcc
     1381 taccctgtac tggcacgtga agaacaagcg ggccctgctc gatgccctgg caatcgagat
     1441 gctggacagg catcataccc acttctgccc cctggaaggc gagtcatggc aagactttct
     1501 gcggaacaac gccaagtcat tccgctgtgc tctcctctca catcgcgacg gggctaaagt
     1561 gcatctcggc acccgcccaa cagagaaaca gtacgaaacc ctggaaaatc agctcgcgtt
     1621 cctgtgtcag caaggcttct ccctggagaa cgcactgtac gctctgtccg ccgtgggcca
     1681 ctttacactg ggctgcgtat tggaggatca ggagcatcaa gtagcaaaag aggaaagaga
     1741 gacacctacc accgattcta tgcccccact tctgagacaa gcaattgagc tgttcgacca
     1801 tcagggagcc gaacctgcct tccttttcgg cctggaacta atcatatgtg gcctggagaa
     1861 acagctaaag tgcgaaagcg gcgggccaaa aaagaagaga aagggaagcg gagctactaa
     1921 cttcagcctg ctgaagcagg ctggagacgt ggaggagaac cctggaccta tggtgagcaa
     1981 gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga
     2041 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     2101 gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga
     2161 catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat
     2221 ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     2281 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
     2341 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa
     2401 gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg
     2461 cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac
     2521 cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt
     2581 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg
     2641 ccgccactcc accggcggca tggacgagct gtacaagggc agcggagcta ctaacttcag
     2701 cctgctgaag caggctggag acgtggagga gaaccctgga cccgggatga tcgagaaaaa
     2761 gaagtccttc gctaagggca tgggcgtgaa gtccacactc gtgtccggct ccaaagtgta
     2821 catgacaacc ttcgccgaag gcagcgacgc caggctggaa aagatcgtgg agggcgacag
     2881 catcaggagc gtgaatgagg gcgaggcctt cagcgctgaa atggccgata aaaacgccgg
     2941 ctataagatc ggcaacgcca aattcagcca tcctaagggc tacgccgtgg tggctaacaa
     3001 ccctctgtat acaggacccg tccagcagga tatgctcggc ctgaaggaaa ctctggaaaa
     3061 gaggtacttc ggcgagagcg ctgatggcaa tgacaatatt tgtatccagg tgatccataa
     3121 catcctggac attgaaaaaa tcctcgccga atacattacc aacgccgcct acgccgtcaa
     3181 caatatctcc ggcctggata aggacattat tggattcggc aagttctcca cagtgtatac
     3241 ctacgacgaa ttcaaagacc ccgagcacca tagggccgct ttcaacaata acgataagct
     3301 catcaacgcc atcaaggccc agtatgacga gttcgacaac ttcctcgata accccagact
     3361 cggctatttc ggccaggcct ttttcagcaa ggagggcaga aattacatca tcaattacgg
     3421 caacgaatgc tatgacattc tggccctcct gagcggactg agacactggg tggtccacaa
     3481 caacgaagaa gagtccagga tctccaggac ctggctctac aacctcgata agaacctcga
     3541 caacgaatac atctccaccc tcaactacct ctacgacagg atcaccaatg agctgaccaa
     3601 ctccttctcc aagaactccg ccgccaacgt gaactatatt gccgaaactc tgggaatcaa
     3661 ccctgccgaa ttcgccgaac aatatttcag attcagcatt atgaaagagc agaaaaacct
     3721 cggattcaat atcaccaagc tcagggaagt gatgctggac aggaaggata tgtccgagat
     3781 caggaaaaat cataaggtgt tcgactccat caggaccaag gtctacacca tgatggactt
     3841 tgtgatttat aggtattaca tcgaagagga tgccaaggtg gctgccgcca ataagtccct
     3901 ccccgataat gagaagtccc tgagcgagaa ggatatcttt gtgattaacc tgaggggctc
     3961 cttcaacgac gaccagaagg atgccctcta ctacgatgaa gctaatagaa tttggagaaa
     4021 gctcgaaaat atcatgcaca acatcaagga atttagggga aacaagacaa gagagtataa
     4081 gaagaaggac gcccctagac tgcccagaat cctgcccgct ggccgtgatg tttccgcctt
     4141 cagcaaactc atgtatgccc tgacaatgtt cctggatggc aaggagatca acgacctcct
     4201 gaccaccctg attaataaat tcgataacat ccagagcttc ctgaaggtga tgcctctcat
     4261 cggagtcaac gctaagttcg tggaggaata cgcctttttc aaagactccg ccaagatcgc
     4321 cgatgagctg aggctgatca agtccttcgc tagaatggga gaacctattg ccgatgccag
     4381 gagggccatg tatatcgacg ccatccgtat tttaggaacc aacctgtcct atgatgagct
     4441 caaggccctc gccgacacct tttccctgga cgagaacgga aacaagctca agaaaggcaa
     4501 gcacggcatg agaaatttca ttattaataa cgtgatcagc aataaaaggt tccactacct
     4561 gatcagatac ggtgatcctg cccacctcca tgagatcgcc aaaaacgagg ccgtggtgaa
     4621 gttcgtgctc ggcaggatcg ctgacatcca gaaaaaacag ggccagaacg gcaagaacca
     4681 gatcgacagg tactacgaaa cttgtatcgg aaaggataag ggcaagagcg tgagcgaaaa
     4741 ggtggacgct ctcacaaaga tcatcaccgg aatgaactac gaccaattcg acaagaaaag
     4801 gagcgtcatt gaggacaccg gcagggaaaa cgccgagagg gagaagttta aaaagatcat
     4861 cagcctgtac ctcaccgtga tctaccacat cctcaagaat attgtcaata tcaacgccag
     4921 gtacgtcatc ggattccatt gcgtcgagcg tgatgctcaa ctgtacaagg agaaaggcta
     4981 cgacatcaat ctcaagaaac tggaagagaa gggattcagc tccgtcacca agctctgcgc
     5041 tggcattgat gaaactgccc ccgataagag aaaggacgtg gaaaaggaga tggctgaaag
     5101 agccaaggag agcattgaca gcctcgagag cgccaacccc aagctgtatg ccaattacat
     5161 caaatacagc gacgagaaga aagccgagga gttcaccagg cagattaaca gggagaaggc
     5221 caaaaccgcc ctgaacgcct acctgaggaa caccaagtgg aatgtgatca tcagggagga
     5281 cctcctgaga attgacaaca agacatgtac cctgttcagg aacaaggccg tccatctgga
     5341 agtggccagg tatgtccacg cctatatcaa cgacattgcc gaggtcaatt cctacttcca
     5401 actgtaccat tacatcatgc agagaattat catgaatgag aggtacgaga aaagcagcgg
     5461 aaaggtgtcc gagtacttcg acgctgtgaa tgacgagaag aagtacaacg ataggctcct
     5521 gaaactgctg tgtgtgcctt tcggctactg tatccccagg tttaagaacc tgagcatcga
     5581 ggccctgttc gataggaacg aggccgccaa gttcgacaag gagaaaaaga aagtgtccgg
     5641 caattccact agtgctccca agaaaaagcg caaggtaggt ggaagcccag cagctaaaag
     5701 agttaaattg gatggatccg attataagga tcacgatgga gattacaagg accacgacat
     5761 agactacaaa gatgatgacg acaagtaacg tacgtaagaa ttcgattcgt cagtagggtt
     5821 gtaaaggttt ttcttttcct gagaaaacaa ccttttgttt tctcaggttt tgctttttgg
     5881 cctttcccta gctttaaaaa aaaaaagcaa aactcaccga ggcagttcca taggatggca
     5941 aggccgcgac tctagagtcg acctgcaggc atgcaagctt gatatcaggc cgcaatcaac
     6001 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     6061 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     6121 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     6181 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     6241 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     6301 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     6361 ctgacaattc cgtggtgttg tcctcgagtc tagagggccc gtttaaaccc gctgatcagc
     6421 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     6481 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     6541 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     6601 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     6661 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     6721 cgcggcgggt gtggtggtta cgcgctaaag caagtaaaac ctctacaaat gtggtatgga
     6781 attctaaaat acagcatagc aaaactttaa cctccaaatc aagcctctac ttgaatcctt
     6841 ttctgaggga tgaataaggc ataggcatca ggggctgttg ccaatgtgca ttagctgttt
     6901 gcagcctcac cttctttcat ggagtttaag atatagtgta ttttcccaag gtttgaacta
     6961 gctcttcatt tctttatgtt ttaaatgcac tgacctccca cattcccttt ttagtaaaat
     7021 attcagaaat aatttaaata catcattgca atgaaaataa atgtttttta ttaggcagaa
     7081 tccagatgct caaggccctt cataatatcc cccagtttag tagttggact tagggaacaa
     7141 aggaaccttt aatagaaatt ggacagcaag aaagcgagct tctagcttta gttcctggtg
     7201 tacttgaggg ggatgagttc ctcaatggtg gttttgacca gcttgccatt catctcaatg
     7261 agcacaaagc agtcaggagc atagtcagag atgagctctc tgcacatgcc acaggggctg
     7321 accaccctga tggatctgtc cacctcatca gagtaggggt gcctgacagc cacaatggtg
     7381 tcaaagtcct tctgcccgtt gctcacagca gacccaatgg caatggcttc agcacagaca
     7441 gtgaccctgc caatgtaggc ctcaatgtgg acagcagaga tgatctcccc agtcttggtc
     7501 ctgatggccg ccccgacatg gtgcttgttg tcctcataga gcatggtgat cttctcagtg
     7561 gcgacctcca ccagctccag atcctgctga gagatgttga aggtcttcat ggtagcctcc
     7621 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     7681 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     7741 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     7801 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     7861 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     7921 ctgacacaca ttccacagct gcgaagagaa gaggtcaggc cgaagttcct atactatttg
     7981 aagaatagga acttcaagct tgccccactg tggggtggag gggacagata aaagtaccct
     8041 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     8101 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     8161 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     8221 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     8281 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     8341 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     8401 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     8461 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     8521 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     8581 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     8641 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     8701 acgatgcccg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     8761 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     8821 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     8881 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     8941 atctacacga cggcgagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     9001 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttattctac ggttctaccg
     9061 ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg
     9121 cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg
     9181 ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag
     9241 gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg
     9301 tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg
     9361 gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg
     9421 tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc
     9481 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc
     9541 ctcatctccg ggcctttcga cctgcagcga cccgcttaac agcgtcaaca gcgtgccgca
     9601 gatcttggtg gcgtgaaact cccgcacctc ttcggcaagc gccttgtaga agcgcgtatg
     9661 gcttcgtacc cctgccatca acacgcgtct gcgttcgacc aggctgcgcg ttctcgcggc
     9721 catagcaacc gacgtacggc gttgcgccct cgccggcagc aagaagccac ggaagtccgc
     9781 ctggagcaga aaatgcccac gctactgcgg gtttatatag acggtcctca cgggatgggg
     9841 aaaaccacca ccacgcaact gctggtggcc ctgggttcgc gcgacgatat cgtctacgta
     9901 cccgagccga tgacttactg gcaggtgctg ggggcttccg agacaatcgc gaacatctac
     9961 accacacaac accgcctcga ccagggtgag atatcggccg gggacgcggc ggtggtaatg
    10021 acaagcgccc agataacaat gggcatgcct tatgccgtga ccgacgccgt tctggctcct
    10081 catatcgggg gggaggctgg gagctcacat gccccgcccc cggccctcac cctcatcttc
    10141 gaccgccatc ccatcgccgc cctcctgtgc tacccggccg cgcgatacct tatgggcagc
    10201 atgacccccc aggccgtgct ggcgttcgtg gccctcatcc cgccgacctt gcccggcaca
    10261 aacatcgtgt tgggggccct tccggaggac agacacatcg accgcctggc caaacgccag
    10321 cgccccggcg agcggcttga cctggctatg ctggccgcga ttcgccgcgt ttacgggctg
    10381 cttgccaata cggtgcggta tctgcagggc ggcgggtcgt ggcgggagga ttggggacag
    10441 ctttcgggga cggccgtgcc gccccagggt gccgagcccc agagcaacgc gggcccacga
    10501 ccccatatcg gggacacgtt atttaccctg tttcgggccc ccgagttgct ggcccccaac
    10561 ggcgacctgt acaacgtgtt tgcctgggcc ttggacgtct tggccaaacg cctccgtccc
    10621 atgcacgtct ttatcctgga ttacgaccaa tcgcccgccg gctgccggga cgccctgctg
    10681 caacttacct ccgggatggt ccagacccac gtcaccaccc ccggctccat accgacgatc
    10741 tgcgacctgg cgcgcacgtt tgcccgggag atgggggagg ctaactgaaa cacggaagga
    10801 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
    10861 gtgttgggtc gtttgttcat aaacgcgctc ataggatctt cttgagatcc tttttttctg
    10921 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
    10981 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
    11041 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
    11101 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
    11161 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
    11221 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
    11281 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
    11341 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
    11401 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
    11461 tgctcgtcag gggggcggag cctatggaaa aacgccgaag ttcctatact ttctagagaa
    11521 taggaacttc acgggccaga tatacgcgac gggccagata tacgcgacgg gccagatata
    11581 cgcgacgggc cagatatacg cgacgggcca gatatacgcg acgggccaga tatacgcgac
    11641 gggccagata tacgcgacgg gccagatata cgcgttgaca ttgattattg actagttatt
    11701 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
    11761 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca t
//
