LOCUS       sequence-401647-        9877 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Encodes a DNase-dead AsCpf1 fused to a NLP nuclear localization 
            signal (NLS) and [Activ] transactivation domain. 3' UTR has a MALAT1
            triplex, dCas12a direct repeats (DR), and a crRNA cloning site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9877)
  TITLE     SGRN: A Cas12a-driven Synthetic Gene Regulatory Network System
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9877)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: 
            pCE059-SiT-ddCas12a(E993A)-1xNLS-[Activ]-triplex-DR-M13-DR-pA
FEATURES             Location/Qualifiers
     source          1..9877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   16..145
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        248..1426
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          479..1417
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1374..1394
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1461..5381
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND
                     HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN
                     AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD
                     KFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF
                     ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL
                     NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL
                     RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG
                     KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL
                     KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA
                     RNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYK
                     ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP
                     LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID
                     LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG
                     HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK
                     DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF
                     HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQR
                     SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV
                     VVLANLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ
                     FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD
                     VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN
                     HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM
                     RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES
                     KDLKLQNGISNQDWLAYIQELRN"
     regulatory      5101..5110
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5382..5429
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5748..6119
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     CDS             6126..6206
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     primer_bind     6403..6419
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6403..6419
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_feature    6480..7068
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6533..6553)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6951..6962)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     repeat_region   7155..7295
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7155..7284
                     /label=AAV2 ITR
     rep_origin      7370..7825
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7457..7476)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7667..7688
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7842..7861)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7961..7983
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8021..8039)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8107..8211
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8212..8280,8281..9072)
                     /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(8430..8449)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9243..9831
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9732..9751
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttttgctca catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg
       61 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
      121 gccaactcca tcactagggg ttcctgcggc cgcacgcgtt taattaatgc aaagatggat
      181 aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct tgaaaggagt
      241 gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa
      301 gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg
      361 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
      421 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt
      481 aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc
      541 ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc gggttggaag
      601 tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga
      661 ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct
      721 cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt
      781 tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt
      841 tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg
      901 cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct
      961 ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc
     1021 ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa
     1081 atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc
     1141 ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca
     1201 cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta
     1261 tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt
     1321 gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc
     1381 tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgagcta gcgtttaaac
     1441 ttaagcttgg taccgccacc atgacacagt tcgagggctt taccaacctg tatcaggtga
     1501 gcaagacact gcggtttgag ctgatcccac agggcaagac cctgaagcac atccaggagc
     1561 agggcttcat cgaggaggac aaggcccgca atgatcacta caaggagctg aagcccatca
     1621 tcgatcggat ctacaagacc tatgccgacc agtgcctgca gctggtgcag ctggattggg
     1681 agaacctgag cgccgccatc gactcctata gaaaggagaa aaccgaggag acaaggaacg
     1741 ccctgatcga ggagcaggcc acatatcgca atgccatcca cgactacttc atcggccgga
     1801 cagacaacct gaccgatgcc atcaataaga gacacgccga gatctacaag ggcctgttca
     1861 aggccgagct gtttaatggc aaggtgctga agcagctggg caccgtgacc acaaccgagc
     1921 acgagaacgc cctgctgcgg agcttcgaca agtttacaac ctacttctcc ggcttttatg
     1981 agaacaggaa gaacgtgttc agcgccgagg atatcagcac agccatccca caccgcatcg
     2041 tgcaggacaa cttccccaag tttaaggaga attgtcacat cttcacacgc ctgatcaccg
     2101 ccgtgcccag cctgcgggag cactttgaga acgtgaagaa ggccatcggc atcttcgtga
     2161 gcacctccat cgaggaggtg ttttccttcc ctttttataa ccagctgctg acacagaccc
     2221 agatcgacct gtataaccag ctgctgggag gaatctctcg ggaggcaggc accgagaaga
     2281 tcaagggcct gaacgaggtg ctgaatctgg ccatccagaa gaatgatgag acagcccaca
     2341 tcatcgcctc cctgccacac agattcatcc ccctgtttaa gcagatcctg tccgatagga
     2401 acaccctgtc tttcatcctg gaggagttta agagcgacga ggaagtgatc cagtccttct
     2461 gcaagtacaa gacactgctg agaaacgaga acgtgctgga gacagccgag gccctgttta
     2521 acgagctgaa cagcatcgac ctgacacaca tcttcatcag ccacaagaag ctggagacaa
     2581 tcagcagcgc cctgtgcgac cactgggata cactgaggaa tgccctgtat gagcggagaa
     2641 tctccgagct gacaggcaag atcaccaagt ctgccaagga gaaggtgcag cgcagcctga
     2701 agcacgagga tatcaacctg caggagatca tctctgccgc aggcaaggag ctgagcgagg
     2761 ccttcaagca gaaaaccagc gagatcctgt cccacgcaca cgccgccctg gatcagccac
     2821 tgcctacaac cctgaagaag caggaggaga aggagatcct gaagtctcag ctggacagcc
     2881 tgctgggcct gtaccacctg ctggactggt ttgccgtgga tgagtccaac gaggtggacc
     2941 ccgagttctc tgcccggctg accggcatca agctggagat ggagccttct ctgagcttct
     3001 acaacaaggc cagaaattat gccaccaaga agccctactc cgtggagaag ttcaagctga
     3061 actttcagat gcctacactg gcctctggct gggacgtgaa taaggagaag aacaatggcg
     3121 ccatcctgtt tgtgaagaac ggcctgtact atctgggcat catgccaaag cagaagggca
     3181 ggtataaggc cctgagcttc gagcccacag agaaaaccag cgagggcttt gataagatgt
     3241 actatgacta cttccctgat gccgccaaga tgatcccaaa gtgcagcacc cagctgaagg
     3301 ccgtgacagc ccactttcag acccacacaa cccccatcct gctgtccaac aatttcatcg
     3361 agcctctgga gatcacaaag gagatctacg acctgaacaa tcctgagaag gagccaaaga
     3421 agtttcagac agcctacgcc aagaaaaccg gcgaccagaa gggctacaga gaggccctgt
     3481 gcaagtggat cgacttcaca agggattttc tgtccaagta taccaagaca acctctatcg
     3541 atctgtctag cctgcggcca tcctctcagt ataaggacct gggcgagtac tatgccgagc
     3601 tgaatcccct gctgtaccac atcagcttcc agagaatcgc cgagaaggag atcatggatg
     3661 ccgtggagac aggcaagctg tacctgttcc agatctataa caaggacttt gccaagggcc
     3721 accacggcaa gcctaatctg cacacactgt attggaccgg cctgttttct ccagagaacc
     3781 tggccaagac aagcatcaag ctgaatggcc aggccgagct gttctaccgc cctaagtcca
     3841 ggatgaagag gatggcacac cggctgggag agaagatgct gaacaagaag ctgaaggatc
     3901 agaaaacccc aatccccgac accctgtacc aggagctgta cgactatgtg aatcacagac
     3961 tgtcccacga cctgtctgat gaggccaggg ccctgctgcc caacgtgatc accaaggagg
     4021 tgtctcacga gatcatcaag gataggcgct ttaccagcga caagttcttt ttccacgtgc
     4081 ctatcacact gaactatcag gccgccaatt ccccatctaa gttcaaccag agggtgaatg
     4141 cctacctgaa ggagcacccc gagacaccta tcatcggcat cgatcggggc gagagaaacc
     4201 tgatctatat cacagtgatc gactccaccg gcaagatcct ggagcagcgg agcctgaaca
     4261 ccatccagca gtttgattac cagaagaagc tggacaacag ggagaaggag agggtggcag
     4321 caaggcaggc ctggtctgtg gtgggcacaa tcaaggatct gaagcagggc tatctgagcc
     4381 aggtcatcca cgagatcgtg gacctgatga tccactacca ggccgtggtg gtgctggcga
     4441 acctgaattt cggctttaag agcaagagga ccggcatcgc cgagaaggcc gtgtaccagc
     4501 agttcgagaa gatgctgatc gataagctga attgcctggt gctgaaggac tatccagcag
     4561 agaaagtggg aggcgtgctg aacccatacc agctgacaga ccagttcacc tcctttgcca
     4621 agatgggcac ccagtctggc ttcctgtttt acgtgcctgc cccatataca tctaagatcg
     4681 atcccctgac cggcttcgtg gaccccttcg tgtggaaaac catcaagaat cacgagagcc
     4741 gcaagcactt cctggagggc ttcgactttc tgcactacga cgtgaaaacc ggcgacttca
     4801 tcctgcactt taagatgaac agaaatctgt ccttccagag gggcctgccc ggctttatgc
     4861 ctgcatggga tatcgtgttc gagaagaacg agacacagtt tgacgccaag ggcacccctt
     4921 tcatcgccgg caagagaatc gtgccagtga tcgagaatca cagattcacc ggcagatacc
     4981 gggacctgta tcctgccaac gagctgatcg ccctgctgga ggagaagggc atcgtgttca
     5041 gggatggctc caacatcctg ccaaagctgc tggagaatga cgattctcac gccatcgaca
     5101 ccatggtggc cctgatccgc agcgtgctgc agatgcggaa ctccaatgcc gccacaggcg
     5161 aggactatat caacagcccc gtgcgcgatc tgaatggcgt gtgcttcgac tcccggtttc
     5221 agaacccaga gtggcccatg gacgccgatg ccaatggcgc ctaccacatc gccctgaagg
     5281 gccagctgct gctgaatcac ctgaaggaga gcaaggatct gaagctgcag aacggcatct
     5341 ccaatcagga ctggctggcc tacatccagg agctgcgcaa caaaaggccg gcggccacga
     5401 aaaaggccgg ccaggcaaaa aagaaaaagg gatccggtgg gggtgggtcc agcgggttgc
     5461 ctaatgggtt ggatggtgac gaggatttta gcgatatcgc cgatatggac ttttcagctc
     5521 tcctcagcca gataagctct ggaagtagtg ggcttccaaa tgggcttgac ggtgacgaag
     5581 atttctctga tatagcggac atggacttct cagctctgct ctcccagatt tcttcagggt
     5641 ccagcggctt gcccaacggc ctcgatggcg acgaggattt ttctgacata gcagacatgg
     5701 acttttccgc actcctcagt cagatttctt ctggaggcgg tggaagcggc ttcagcgtgg
     5761 acaccagtgc cctgctggac ctgttcagcc cctcggtgac cgtgcccgac atgagcctgc
     5821 ctgaccttga cagcagcctg gccagtatcc aagagctcct gtctccccag gagcccccca
     5881 ggcctcccga ggcagagaac agcagcccgg attcagggaa gcagctggtg cactacacag
     5941 cgcagccgct gttcctgctg gaccccggct ccgtggacac cgggagcaac gacctgccgg
     6001 tgctgtttga gctgggagag ggctcctact tctccgaagg ggacggcttc gccgaggacc
     6061 ccaccatctc cctgctgaca ggctcggagc ctcccaaagc caaggacccc actgtctccg
     6121 gatcctaccc atacgatgtt ccagattacg cttatcccta cgacgtgcct gattatgcat
     6181 acccatatga tgtccccgac tatgcctaag aattcgattc gtcagtaggg ttgtaaaggt
     6241 ttttcttttc ctgagaaaac aaccttttgt tttctcaggt tttgcttttt ggcctttccc
     6301 tagctttaaa aaaaaaaaag caaaactcac cgaggcagtt ccataggatg gcaagatcct
     6361 ggtattggtc tgcgaaattt ctactcttgt agattgaaga gccaggaaac agctatgacg
     6421 ctcttctaat ttctactctt gtagattcta gaaaagaatt cgatatcaag cttatcgata
     6481 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     6541 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     6601 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     6661 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     6721 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     6781 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     6841 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     6901 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     6961 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     7021 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag
     7081 cgctaataaa agatctttat tttcattaga tctgtgtgtt ggttttttgt gtcacgtgcg
     7141 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     7201 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     7261 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     7321 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     7381 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     7441 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     7501 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     7561 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     7621 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     7681 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     7741 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     7801 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     7861 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     7921 ctgctcccgg catccgctta cagacaagct gtgaccttct ccgggagctg catgtgtcag
     7981 aggttttcac cgtcatcacc gaaacgcgcg atacgaaagg gcctcgtgat acgcctattt
     8041 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     8101 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     8161 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     8221 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     8281 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     8341 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     8401 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     8461 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     8521 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     8581 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     8641 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     8701 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     8761 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     8821 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     8881 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     8941 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     9001 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     9061 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     9121 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     9181 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     9241 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     9301 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     9361 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     9421 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     9481 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     9541 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     9601 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     9661 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     9721 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     9781 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     9841 aacgcggcct ttttacggtt cctggccttt tgctggc
//
