LOCUS       sequence-261581-       10030 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Swap out effector and terminator to generate SPARC2-D CRISPR-donor 
            vector for CRISPR-HDR genomic insertion near the attP40 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10030)
  AUTHORS   Isaacman-Beck J, Paik KC, Wienecke CFR, Yang HH, Fisher YE, Wang IE,
            Ishida IG, Maimon G, Wilson RI, Clandinin TR
  TITLE     SPARC enables genetic manipulation of precise proportions of cells.
  JOURNAL   Nat Neurosci. 2020 Sep;23(9):1168-1175. doi: 
            10.1038/s41593-020-0668-9. Epub 2020 Jul 20.
  PUBMED    32690967
REFERENCE   2  (bases 1 to 10030)
  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: 
            "10.1038/s41593-020-0668-9"; journalName: "Nat Neurosci"; date: 
            "2020-09"; volume: "23"; issue: "9"; pages: "1168-1175"
COMMENT     Sequence Label: pHD-SPARC2-D-LexA::p65
FEATURES             Location/Qualifiers
     source          1..10030
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     622..641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(860..951)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     993..1012
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1018..1037
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    2348..2442
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    2476..2570
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    2604..2698
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    2732..2826
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     promoter        2851..3089
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     3036..3057
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     intron          3325..3390
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3520..3540
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3812..3946
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    4349..4418
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     CDS             join(4425..4676,4677..5030)
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5'-CTGTATATATATACAG-3'."
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     CDS             5475..5846
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     protein_bind    6150..6183
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      6409..6418
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6415..7092
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(6545..6563)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(6595..6615)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     6813..6832
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7007..7030
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    7213..7334
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    7341..7374
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(9033..9050)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9033..9049)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9148..9165)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9319..9907)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9399..9418)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10029..10030,1..790,791..859))
                     /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"
ORIGIN
        1 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
       61 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
      121 gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca gcaataaacc
      181 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
      241 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
      301 ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
      361 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
      421 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
      481 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
      541 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
      601 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
      661 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
      721 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
      781 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
      841 ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt tatcagggtt
      901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggtca
      961 gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata cctgaatatg
     1021 gctcataaca ccccttgttt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     1081 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggactcc ccatgcgaga
     1141 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     1201 cccgggctaa ttatggggtg tcgcccttcg cacagaaaag catcttacgg atggtgtttg
     1261 ccaaaagtgc tggagatgtt ttagatatag atggtttatt ttaagaaatg gatgtatata
     1321 attgagctac tgcatcacgg gtttctgttt aacagtaaca gtgaaaagtg ctcaatgata
     1381 catatattac aaataataac ttttggtttt taaagtagat ttatataata taatcataat
     1441 tcttaatggg aatccatgtt acgaccaaat tagccgtggc ctttcaagtg gctgccactt
     1501 atattttaca actcgcacac agtggctcac accctcgtaa acaaaataat gctattaaat
     1561 caaattgaac atgtgaattc tctcaagaag acgccacttt tttttggggg gccagctgca
     1621 actgccacca atgtttactt ttatctttaa tgggcacatg ccttacatat acgaatttta
     1681 cacgattttt caagtggcgg tcgagagtgg ggtgataatt aattaaggtt gcctttagat
     1741 aaagtattat agagtttcaa aaaaatgaaa cttaagaatg gaatagctgc attccattct
     1801 atgtagattt cccttcaaaa ttattaaaat ccgttgacct ttgttcattt agcaattaga
     1861 attcgtctat gaataacaaa ttaaattgcg ttgcgggcaa attgctttag gccatttgaa
     1921 gtgcaacttt atatctcagg gccaacttct cggcgcgaca acaataaaaa catatacata
     1981 gatacacata tcagatgcaa tgcaatcacc acagcgtgac gtcattagtc taggcttgaa
     2041 aatacacggc aaacttgttt ataactctac gctgattaca taagccgcaa acacgacgaa
     2101 aaaacatttg ttttggtttc aaatccatga atgactttca ttaagtgcag ctgtaagctg
     2161 agctttgcca tgcgctgacc catattttca gcctctcgtt ctctctgttt cgcagatact
     2221 gtgaatatta gtaaaacgat ccataggggc tgtagtttat aatcataatc aaataggcat
     2281 aaacaaccac cagcgcggcc gcggacatat gggatccagt gcaccgtaag cttgcatgcc
     2341 tgcaggtcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc
     2401 cgagcggagt actgtcctcc gagcggagta ctgtcctccg agcggagact ctagccctag
     2461 ggcatgcctg caggtcggag tactgtcctc cgagcggagt actgtcctcc gagcggagta
     2521 ctgtcctccg agcggagtac tgtcctccga gcggagtact gtcctccgag cggagactct
     2581 agcgctagcg catgcctgca ggtcggagta ctgtcctccg agcggagtac tgtcctccga
     2641 gcggagtact gtcctccgag cggagtactg tcctccgagc ggagtactgt cctccgagcg
     2701 gagactctag cactagtgca tgcctgcagg tcggagtact gtcctccgag cggagtactg
     2761 tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg agtactgtcc
     2821 tccgagcgga gactctagcg acgtcgagcg ccggagtata aatagaggcg cttcgtctac
     2881 ggagcgacaa ttcaattcaa acaagcaaag tgaacacgtc gctaagcgaa agctaagcaa
     2941 ataaacaagc gcagctgaac aagctaaaca atctgcagta aagtgcaagt taaagtgaat
     3001 caattaaaag taaccagcaa ccaagtaaat caactgcaac tactgaaatc tgccaagaag
     3061 taattattga atacaagaag agaactctga attaacgtaa gctagctaga ggatctttgt
     3121 gaaggaacct tacggtacca atcaaaagat ctactccagt cgggagtagt gccccaactg
     3181 gggtaacctt tgagttctct cagttggggg cgtagggtcg taacgtcgta agctagctag
     3241 aggatctttg tgaaggaacc ttacttggta ccctgtggtg tgacataatt ggacaaacta
     3301 cctacagaga tttaaagctc taaggtaaat ataaaatttt taagtgtata atgtgttaaa
     3361 ctactgattc taattgtttg tgtattttag attccaacct atggaactga tgaatgggag
     3421 cagtggtgga atgcctttaa tgaggaaaac ctgttttgct cagaagaaat gccatctagt
     3481 gatgatgagg ctactgctga ctctcaacat tctactcctc caaaaaagaa gagaaaggta
     3541 gaagacccca aggactttcc ttcagaattg ctaagttttt tgagtcatgc tgtgtttagt
     3601 aatagaactc ttgcttgctt tgctatttac accacaaagg aaaaagctgc actgctatac
     3661 aagaaaatta tggaaaaata tttgatgtat agtgccttga ctagagatca taatcagcca
     3721 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3781 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     3841 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3901 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcgatc tggccggcgg
     3961 gagtagtgcc ccaactgggg taacctttga gttctctcag ttgggggcgt agggtcgggg
     4021 tcggcatggc atctccacct cctcgcggtc cgacctgggc atccgaagga ggacgtcgtc
     4081 cactcggatg gctaagggag agctcggatc cgggcaatac gaggtactgg cagacacgca
     4141 gtgtcgaaca acacctgacc taacggtaag atgagtagat aactcaacta cttaaatcct
     4201 ggcctgcttg atatctcgta gcgttcttag agatcgacga actgtttcac gaccgggtcg
     4261 gcatggcatc tccacctcct cgcggtccga cctgggcatc cgaaggagga cgtcgtccac
     4321 tcggatggct aagggagagc tcggatccct cgaagccgcg gtgcgggtgc cagggcgtgc
     4381 ccttgggctc cccgggcgcg tactccacct cacccatcgt cgacatgaag gctctcacgg
     4441 cccgacaaca ggaagttttt gatttgatac gggatcatat atcccaaacg ggtatgcctc
     4501 cgacccgcgc agagatagca cagcgactgg gctttcgatc gcctaacgcc gcggaggagc
     4561 acttgaaggc actggcccgc aagggtgtca ttgaaatcgt gtccggtgcg agccgcggaa
     4621 tccggctgtt gcaggaggag gaagagggcc tgccactggt gggacgcgtg gccgctggcg
     4681 agccgctgct ggcccagcaa cacatagagg gacactatca ggtggacccc tccttgttta
     4741 agccaaatgc tgatttcctg ttgcgggtgt cgggaatgtc catgaaggac atcggtatta
     4801 tggatggtga cctcctcgcc gtccataaga cacaggatgt caggaacggc caggtagtcg
     4861 ttgccaggat agacgatgag gtcactgtga aacgtctcaa gaagcaaggc aataaggtcg
     4921 agctgctgcc ggagaatagc gagttcaagc cgatcgtggt ggatctgcga cagcagtcct
     4981 ttactatcga gggcttggcc gtgggtgtga tccgcaacgg agattggctg ggatccacgc
     5041 cgatggagtt ccagtacctg cccgatacgg atgaccgtca ccgtatcgaa gaaaagcgga
     5101 agcgaaccta tgaaaccttc aagtccatca tgaaaaagtc ccccttctcg ggccccacgg
     5161 acccgcgccc cccgccccgt cgtattgcgg ttccttcgcg cagcagtgcc agcgtcccca
     5221 aacccgcacc gcagccctac ccgttcactt cctcccttag cacgattaac tatgatgagt
     5281 tccccacgat ggtgttcccc agtggacaaa tttcccaggc atcggcactg gctccggccc
     5341 caccgcaagt cctcccccag gcgcccgctc cggcaccggc tcccgcaatg gtgagtgctc
     5401 tggcccaggc ccccgctcca gtccccgtgc tggcgcctgg acccccacag gcagttgccc
     5461 ctcctgctcc gaaaccaacg caggcgggcg aaggaaccct gagcgaggcc ctcttgcagc
     5521 ttcagttcga tgacgaagac ttgggagccc tgctgggtaa cagcacagac cctgccgtat
     5581 tcaccgatct cgcatccgtg gacaacagcg agtttcagca gctcttgaat cagggaatcc
     5641 cggtcgcacc tcataccaca gagcccatgc tgatggaata cccggaggct atcacgcgac
     5701 tggtgaccgg cgcacagcga ccacccgatc cagcccctgc cccactgggt gccccgggtt
     5761 tgcccaatgg cctcctcagc ggcgatgagg atttctccag catcgctgat atggatttct
     5821 ccgctttgct gagccagata agctcctaag tttaaacaag cttatcgata ccgtcgacta
     5881 aagccaaata gaaaattatt cagttcctgg cttaagtttt taaaagtgat attatttatt
     5941 tggttgtaac caaccaaaag aatgtaaata actaatacat aattatgtta gttttaagtt
     6001 agcaacaaat tgattttagc tatattagct acttggttaa taaatagaat atatttattt
     6061 aaagataatt gcgtttttat tgtcagggag tgagtttgct taaaaactcg tttagatcca
     6121 ctaggtcgac tgaggcactg cctcgagaga taacttcgta taatgtatgc tatacgaagt
     6181 tatcgtacgg gatctaattc aattagagac taattcaatt agagctaatt caattaggat
     6241 ccaagcttat cgatttcgaa ccctcgaccg ccggagtata aatagaggcg cttcgtctac
     6301 ggagcgacaa ttcaattcaa acaagcaaag tgaacacgtc gctaagcgaa agctaagcaa
     6361 ataaacaagc gcagctgaac aagctaaaca atcggctcga agccggtcgc caccatggcc
     6421 tcctccgagg acgtcatcaa ggagttcatg cgcttcaagg tgcgcatgga gggctccgtg
     6481 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     6541 accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
     6601 ccccagttcc agtacggctc caaggtgtac gtgaagcacc ccgccgacat ccccgactac
     6661 aagaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     6721 ggcgtggtga ccgtgaccca ggactcctcc ctccaggacg gctccttcat ctacaaggtg
     6781 aagttcatcg gcgtgaactt cccctccgac ggccccgtaa tgcagaagaa gactatgggc
     6841 tgggaggcgt ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagatccac
     6901 aaggccctga agctgaagga cggcggccac tacctggtgg agttcaagtc catctacatg
     6961 gccaagaagc ccgtgcagct gcccggctac tactacgtgg actccaagct ggacatcacc
     7021 tcccacaacg aggactacac catcgtggag cagtacgagc gcgccgaggg ccgccaccac
     7081 ctgttcctgt aggggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt
     7141 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     7201 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     7261 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     7321 atcaatgtat cttaaccggt ataacttcgt ataatgtatg ctatacgaag ttatcacatt
     7381 gggtagcaca gactctgagc tttactacga ttactataca gctcttctct cgcgactttt
     7441 tggactggac aaggcgtagc acattgaacg gcagtgggtt tgggtttagt atcgaaccgg
     7501 ctttctacga cagcggattg gaagcgcgga gcgacaaagt cgccgggtaa taatgtgata
     7561 tagccggcta tgttcagcag gcaaaactga aataaaagta ataaacaccg aaagcccccg
     7621 aattctcgca tcttcttttg tgggactgcg agaagaattg cgagcgaaat actaatttcc
     7681 ttacctaact gtgtgagata atctccggtt ggcattgtgt gtcgcgcaat aaaatcccca
     7741 aaagcccaaa gcttcaagtg tttaatataa taaaatagcc aagaagcgtg tgtatttgca
     7801 tttgggaaaa atttaaggtt gaattacgtg ggccgaaaaa ttaaagactg aaggccattc
     7861 aagggttgtt caaagagaaa attcggtgaa aagtcaccgt gtgtgtgtgt gtgagtgtgt
     7921 gtgcgttgcg tgagaaaaat caattctgca tttggtggaa agccgaaaac ggagagaaaa
     7981 ttggtggagc gaggagaaaa ggacacgaat taaaggctgg ttttttgggg ctgcgaaaaa
     8041 ctcgtgaccg atgacaattt ggcttgacga gggtctatta gcgaaatcaa tgctattatg
     8101 gctgatagtg gaggaccagg atccaagcac atggatccaa caaccataga tgccggcggg
     8161 ggcggagcag gagcagccgg aggtgatgac gtgcctccag tcccagctgt tcgccggcgt
     8221 cgtgcgcatg agcagaaatc ctcgagagag caggtgctgg aggaggagaa gagccaacag
     8281 ctggagacat cgacgatgac gaggacctac atgaagacca tcaccacctc cctcaccacc
     8341 tcgagtagct ccaacgtgga agagtttatc ctgggggagc acgcagcagg agcagcagct
     8401 gctccatcgc ccaaccagca gcgattgcgt caaaaggtgg cccagtacga gaaggtctgg
     8461 tcggatggct ccagtcccgt taagcgtcca gccgagcaga gctctgatga gctgcgtctg
     8521 accgacgacc aggatgagta cgccatccgt aagatgcttt tctgtgtcaa tcgagttcaa
     8581 gggcgacaca aaatttattc taaatgcata ataaatactg ataacatctt atagtttgta
     8641 ttatattttg tattatcgtt gacatgtata attttgatat caaaaactga ttttcccttt
     8701 attattttcg agatttattt tcttaattct ctttaacaaa ctagaaatat tgtatataca
     8761 aaaaatcata aataatagat gaatagttta attataggtg ttcatcaatc gaaaaagcaa
     8821 cgtatcttat ttaaagtgcg ttgctttttt ctcatttata aggttaaata attctcatat
     8881 atcaagcaaa gtgacaggcg cccttaaata ttctgacaaa tgctctttcc ctaaactccc
     8941 cccataaaaa aacccgccga agcgggtttt tacgttattt gcggattaac gattactcgt
     9001 tatcagaacc gcccaggggg cccgagctta agactggccg tcgttttaca acacagaaag
     9061 agtttgtaga aacgcaaaaa ggccatccgt caggggcctt ctgcttagtt tgatgcctgg
     9121 cagttcccta ctctcgcctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     9181 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     9241 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     9301 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     9361 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     9421 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     9481 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     9541 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     9601 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     9661 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     9721 gttcttgaag tggtgggcta actacggcta cactagaaga acagtatttg gtatctgcgc
     9781 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     9841 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     9901 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgacgcgcg
     9961 cgtaactcac gttaagggat tttggtcatg agcttgcgcc gtcccgtcaa gtcagcgtaa
    10021 tgctctgctt
//
