LOCUS       sequence-478429-        9090 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Linker-Cno_mNeonGreen-CBP-2xFLAG-CnU6-sgRNA-amdS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9090)
  AUTHORS   Nalley MJ, Banerjee S, Huang MY, Madhani HD
  TITLE     Near 100% efficient homology-dependent genome engineering in the 
            human fungal pathogen Cryptococcus neoformans
  JOURNAL   bioRxiv 2025.02.14.638277
REFERENCE   2  (bases 1 to 9090)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2025.02.14.638277"
COMMENT     Sequence Label: pBHM2662
FEATURES             Location/Qualifiers
     source          1..9090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     1058..1074
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1059..1075
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1093..1116
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     regulatory      1381..1390
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1846..1923
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             1936..1983
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     misc_RNA        complement(2323..2398)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(2922..4568)
                     /codon_start=1
                     /gene="amdS"
                     /product="acetamidase from Aspergillus nidulans"
                     /label=amdS
                     /note="confers ability to process acetamide for selection 
                     in fungal cells"
                     /translation="MPQSWEELAADKRARLAKTIPDEWKVQTLPAEDSVIDFPKKSGIL
                     SEAELKITEASAADLVSKLAAGELTSVEVTLAFCKRAAIAQQLTNCAHEFFPDAALAQA
                     RELDEYYAKHKRPVGPLHGLPISLKDQLRVKGYETSMGYISWLNKYDEGDSVLTTMLRK
                     AGAVFYVKTSVPQTLMVCETVNNIIGRTVNPRNKNWSCGGSSGGEGAIVGIRGGVIGVG
                     TDIGGSIRVPAAFNFLYGLRPSHGRLPYAKMANSMEGQETVHSVVGPITHSVEDLRLFT
                     KSVLGQEPWKYDSKVIPMPWRQSESDIIASKIKNGGLNIGYYNFDGNVLPHPPILRGVE
                     TTVAALAKAGHTVTPWTPYKHDFGHDLISHIYAADGSADVMRDISASGEPAIPNIKDLL
                     NPNIKAVNMNELWDTHLQKWNYQMEYLEKWREAEEKAGKELDAIIAPITPTAAVRHDQF
                     RYYGYASVINLLDFTSVVVPVTFADKNIDKKNESFKAVSELDALVQEEYDPEAYHGAPV
                     AVQVIGRRLSEERTLAIAEEVGKLLGNVVTP"
     primer_bind     complement(5365..5384)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5366..5384)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5391..5408)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5391..5407)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5400..5422)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5548..6003
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5635..5654)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5845..5866
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6134..6937)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(6938..7153)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7177..7196)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7296..7318
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7356..7374)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    7415..7918
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        7955..8059
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8060..8128,8129..8920)
                     /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(8278..8297)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg aaattaaccc
     1021 tcactaaagg gaacaaaagc tggtaccggg ccccccctcg aggtcgacgg tatcgataag
     1081 cttggtggcg gactcgaagt cttgttccaa ggtcctggat ctggtatggt ctccaagggt
     1141 gaggaggaca acatggcctc cctccccgcc acccacgagc tccacatctt cggttccatc
     1201 aacggtgtcg acttcgacat ggtcggtcag ggtaccggta accccaacga cggttacgag
     1261 gagctcaacc tcaagtccac caagggtgac ctccagttct ccccctggat cctcgtcccc
     1321 cacatcggtt acggtttcca ccagtacctc ccctaccccg acggtatgtc ccccttccag
     1381 gccgccatgg tcgacggttc cggttaccag gtccaccgaa ccatgcagtt cgaggacggt
     1441 gcctccctca ccgtcaacta ccgatacacc tacgagggtt cccacatcaa gggtgaggcc
     1501 caggtcaagg gtaccggttt ccccgccgac ggtcccgtca tgaccaactc cctcaccgcc
     1561 gccgactggt gtcgatccaa gaagacctac cccaacgaca agaccatcat ctccaccttc
     1621 aagtggtcct acaccaccgg taacggtaag cgataccgat ccaccgcccg aaccacctac
     1681 accttcgcca agcccatggc cgccaactac ctcaagaacc agcccatgta cgtcttccga
     1741 aagaccgagc tcaagcactc caagaccgag ctcaacttca aggagtggca gaaggccttc
     1801 accgacgtca tgggtatgga cgagctctac aaggggtcag ggtcaaagag aagatggaaa
     1861 aagaatttca tagccgtctc agcagccaac cgctttaaga aaatctcatc ctccggggca
     1921 cttgggtcag ggtcagatta taaagatgac gatgacaagg attataaaga tgacgatgac
     1981 aagtaatctg gacatgtcaa ttgtacactg ccttataatg ccccatgcac ttccagtatt
     2041 agtcatttga agcgttgaaa gatctcttgg cagtgtgatc atggaaacaa tcccaaacag
     2101 cagcaaactg aaaacgttct ggtatgcaat ccatcccatc gtacaagctc tccaaatacc
     2161 tcttccaatc caaaaacggt ttagatgaaa gtctcgtctg atgcacttgt tagtgacacc
     2221 ccagaatttg atgatcttcg actcacctcg agcggccgcc agtgtgatgg atatctgcag
     2281 aattcgccct tgaatagagt cacttttgct agggataaaa aagcaccgac tcggtgccac
     2341 tttttcaagt tgataacgga ctagccttat tttaacttgc tatttctagc tctaaaacct
     2401 aggcatgagg atatctcgca tctagcaaca gtataccctg ccggtgtatg tgcaatttcc
     2461 actttggcgg ataacggaag caaataagac ggcggtccac gccgtctcag tgttttttta
     2521 tttttttccc gtcatccatt gtttttctcc tcacctcttc attctcaaca aatggttata
     2581 attatcggtc acacctgcag gttaccattg tccttgcacg tatactttgc atgcatgccg
     2641 atcaatttaa aaaataaatg aactgtaata atcatgcttt gtttttagtt ctaatgcaag
     2701 ggggatccac tagttgccat gaagatcctg aggattcaac ggcgttgctg agggtgggaa
     2761 ggatggtatt gttgtctgta gatagattaa tgggcatata atgtcaaaaa agatgattga
     2821 cacgatgtct gtctcgaatt cgcagtgtgc taaaattgta tcacttctct aatatgtata
     2881 tatacaccct ctaaggaaaa ttaacccctt accgccttca cctatggagt caccacattt
     2941 cccagcaact tccccacttc ctctgcaatc gccaacgtcc tctcttcact gagtctccgt
     3001 ccgataacct gcactgcaac cggtgcccca tggtacgcct ccggatcata ctcttcctgc
     3061 acgagggcat caagctcact aaccgccttg aaactctcat tcttcttatc gatgttctta
     3121 tccgcaaagg taaccggaac aaccacgctc gtgaaatcca gcaggttgat cacagaggca
     3181 tacccatagt accggaactg gtcatgccgt accgcagcgg taggcgtaat cggcgcgatg
     3241 atggcgtcca gttccttccc ggccttttct tcagcctccc gccatttctc aaggtactcc
     3301 atctggtaat tccacttctg gagatgcgtg tcccagagct cgttcatgtt aacagctttg
     3361 atgttcgggt tcagtaggtc tttgatattt ggaatcgccg gctcgccgga tgcactgata
     3421 tcgcgcatta cgtcggcgct gccgtcagcc gcgtagatat gggagatgag atcgtggccg
     3481 aaatcgtgct tgtatggcgt ccacggggtc acggtgtgac cggctttggc gagtgcggcg
     3541 acggtggttt ccacgccgcg caggatagga gggtgtggaa ggacattgcc gtcgaagttg
     3601 tagtagccga tattgagccc gccgttcttg atcttggagg caataatgtc cgactcggac
     3661 tggcgccagg gcatggggat gaccttggag tcgtatttcc atggctcctg accgaggacg
     3721 gatttggtga agaggcggag gtcctcaaca gagtgcgtaa tcggcccgac aacgctgtgc
     3781 accgtctcct gaccctccat gctgttcgcc atctttgcat acggcagccg cccatgactc
     3841 ggccttagac cgtacaggaa gttgaacgcg gccggcactc gaatcgagcc accgatatcc
     3901 gttcctacac cgatgacgcc accacgaatc ccaacgatcg caccctcacc accagaactg
     3961 ccgccgcacg accagttctt gttgcgtggg ttgacggtgc gcccgatgat gttgttgact
     4021 gtctcgcaga ccatcagggt ctgcgggaca gaggtcttga cgtagaagac ggcaccggct
     4081 ttgcggagca tggttgtcag aaccgagtcc ccttcgtcgt acttgtttag ccatgagatg
     4141 tagcccattg atgtttcgta gcccttgact cgaagctggt ctttgagaga gatggggagg
     4201 ccatggagtg gaccaacggg tctcttgtgc tttgcgtagt attcatcgag ttcccttgcc
     4261 tgcgcgagag cggcgtcagg gaagaactcg tgggcgcagt ttgttaactg ctgggcgatt
     4321 gctgcccgtt tacagaatgc tagcgtaact tccaccgagg tcaactctcc ggccgccagc
     4381 ttggacacaa gatctgcagc ggaggcctct gtgatcttca gttcggcctc tgaaaggatc
     4441 cccgatttct ttgggaaatc aataacgctg tcttccgcag gcagcgtctg gactttccat
     4501 tcatcaggga tggtttttgc gaggcgggcg cgcttatcag cggccagttc ttcccaggat
     4561 tgaggcatag acatgttggg cgagtttact aatggaaaaa gacaattagc aaagaggctg
     4621 catattagga gacgcggtga agattgggga ggacaatagg gtccgtgcac caaaaagtcc
     4681 tgacggcagc tctgttcaac gcacatccat ctcttccagc tatgcgtgtg gcaccagcac
     4741 ggatggattg ttgcaccaaa aggacaaggc gagatggatg caggtcgagg cgatgcggtg
     4801 ctatcggccc tcgttgtcga acgccacgtc gtcctctcgg cgaccgtcaa tcctggaagg
     4861 aaatgaccaa gcgtgcaccc agcatccagc atccaacatc cagcacccag cacctagcac
     4921 ccatgatcca atggccggca tgcagatcag cgtggggaga gcaccatcta gcagcccctg
     4981 gcaggcccag cattgtgctc ggatgtgcga taggcggtgg gtgcgataag gtggattagt
     5041 ggaggatgga ggactgagtg gcgggcaagg gaagggggcg ggaagatgac tcaccttggg
     5101 cagtgggtgg gggatgcggt ataggggatg tatgtggatg ctggcggagg atagaagctg
     5161 taagctgagc tggtgccaac ggtgacgctg tgagagtggt ctcggcgacc ctgacggcga
     5221 ctcggcgtta ccccgtgtgg cacaccggac gcgcgtcgag tgacgttgct gctattgttg
     5281 ttaccatcat cctctcctcc gacaaccata cactcagtgc acaccatgcc actcgaatcc
     5341 tgcatgcggt ggagctccaa ttcgccctat agtgagtcgt attacaattc actggccgtc
     5401 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     5461 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     5521 cagttgcgca gcctgaatgg cgaatggacg cgccctgtag cggcgcatta agcgcggcgg
     5581 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     5641 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     5701 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     5761 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5821 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     5881 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     5941 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     6001 tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagggta
     6061 ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt
     6121 ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt
     6181 ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa
     6241 caataataat gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca
     6301 atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt
     6361 catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct
     6421 tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca
     6481 attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca
     6541 aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg
     6601 ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt
     6661 ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg
     6721 tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg
     6781 gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac
     6841 acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat
     6901 gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg
     6961 tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac
     7021 atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg
     7081 agattaccga atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa
     7141 atgatgaatt gaaaaggtgg tatggtgcac tctcagtaca atctgctctg atgccgcata
     7201 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     7261 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     7321 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     7381 ggttaatgtc atgataataa tggtttctta gacggatcgc ttgcctgtaa cttacacgcg
     7441 cctcgtatct tttaatgatg gaataatttg ggaatttact ctgtgtttat ttatttttat
     7501 gttttgtatt tggattttag aaagtaaata aagaaggtag aagagttacg gaatgaagaa
     7561 aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc gtactttaca tatatattta
     7621 ttagacaaga aaagcagatt aaatagatat acattcgatt aacgataagt aaaatgtaaa
     7681 atcacaggat tttcgtgtgt ggtcttctac acagacaaga tgaaacaatt cggcattaat
     7741 acctgagagc aggaagagca agataaaagg tagtatttgt tggcgatccc cctagagtct
     7801 tttacatctt cggaaaacaa aaactatttt ttctttaatt tcttttttta ctttctattt
     7861 ttaatttata tatttatatt aaaaaattta aattataatt atttttatag cacgtgatga
     7921 aaaggaccca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     7981 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     8041 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     8101 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     8161 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     8221 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     8281 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     8341 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     8401 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     8461 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     8521 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     8581 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     8641 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     8701 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     8761 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     8821 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     8881 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     8941 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     9001 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     9061 agaccccgta gaaaagatca aaggatcttc
//
