LOCUS       sequence-256344-        9961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Piggybac vector for shRNA-mediated knockdown, co-expressed with 
            FUCCI cell cycle reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9961)
  AUTHORS   Fischer AD, Veronese-Paniagua DA, Swaminathan S, Kashima H, Rubin 
            DC, Madison BB
  TITLE     The oncogenic function of PLAGL2 is mediated via ASCL2 and IGF2 and 
            a Wnt-independent mechanism in colorectal cancer.
  JOURNAL   Am J Physiol Gastrointest Liver Physiol. 2023 Jun 13. doi: 
            10.1152/ajpgi.00058.2022.
  PUBMED    37310750
REFERENCE   2  (bases 1 to 9961)
  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"; journalName: "Am J 
            Physiol Gastrointest Liver Physiol. 2023 Jun 13. doi: 
            10.1152/ajpgi.00058.2022."
COMMENT     Sequence Label: BII-Sh-FUCCI
FEATURES             Location/Qualifiers
     source          1..9961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..39
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(308..324)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(308..324)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        580..828
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     580..600
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     751..770
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     CDS             2175..2228
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2235..2261
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             2235..2261
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(2268..2270,2271..2273,2274..2984)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from GFP (Lam et al., 2012)"
                     /label=Clover
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2313..2334)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2921..2942
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3015..3344
                     /codon_start=1
                     /product="degron consisting of residues 1-110 of human 
                     Geminin (Zielke and Edgar, 2015)"
                     /label=Gem1 (1-110)
                     /translation="MNPSMKQKQEEIKENIKNSSVPRRTLKMIQPSASGSLVGRENELS
                     AGLSKRKHRNDHLTSTTSSPGVIVPESSENKNLGGVTQESFDLMIKENPSSQYWKEVAE
                     KRRKAL"
     misc_feature    3349..3921
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3516..3533)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3743..3762
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3936..4586
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAH"
     CDS             3936..4586
                     /codon_start=1
                     /product="humanized monomeric Kusabira-Orange fluorescent 
                     protein"
                     /label=hmKO
                     /note="mammalian codon-optimized"
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAH"
     CDS             4641..4913
                     /codon_start=1
                     /product="degron consisting of residues 30-120 of human 
                     Cdt1 (Zielke and Edgar, 2015)"
                     /label=Cdt1 (30-120)
                     /translation="PSPARPALRAPASATSGSRKRARPPAAPGRDQARPPARRRLRLSV
                     DEVSSPSTPEAPDIPACPSPGQKIKKSTPAAGQPPHLTSAQDQDTI"
     misc_feature    4938..5526
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4991..5011)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5409..5420)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5606..5740
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5630..5649)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5684..5703
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   6518..6580
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6710..6727)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6710..6726)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6719..6741)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6868..7323
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6955..6974)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7165..7186
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     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"
     rep_origin      8485..9073
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8974..8993
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9227..9244
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9361..9382
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9397..9402,9403..9420,9421..9427)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9435..9451
                     /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     9440..9462
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9459..9475
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9459..9475
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttaaccctag aaagatagtc tgcgtaaaat tgacgcatgc attcttgaaa tattgctctc
       61 tctttctaaa tagcgcgaat ccgtcgctgt gcatttagga catctcagtc gccgcttgga
      121 gctcccgtga ggcgtgcttg tcaatgcggt aagtgtcact gattttgaac tataacgacc
      181 gcgtgagtca aaatgacgca tgattatctt ttacgtgact tttaagattt aactcatacg
      241 ataattatat tgttatttca tgttctactt acgtgataac ttattatata tatattttct
      301 tgttatagat ccactagttc tagagggaca gccccccccc aaagccccca gggatgtaat
      361 tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct ccggtccggc
      421 gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg gggaaggtgg
      481 cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag acacctgggg
      541 ggatacgggg aaaaggcctc cagggccgcc cccttcaccg agggcctatt tcccatgatt
      601 ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat taatttgact
      661 gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag
      721 tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag
      781 tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg gagacgctcg
      841 acccgtctcg tttttgcctc caaggcctgc aggagtcaat gggaaaaacc cattggagcc
      901 aagtacactg actcaatagg gactttccat tgggttttgc ccagtacata aggtcaatag
      961 ggggtgagtc aacaggaaag tcccattgga gccaagtaca ttgagtcaat agggactttc
     1021 caatgggttt tgcccagtac ataaggtcaa tgggaggtaa gccaatgggt ttttcccatt
     1081 actgacatgt atactgagtc attagggact ttccaatggg ttttgcccag tacataaggt
     1141 caataggggt gaatcaacag gaaagtccca ttggagccaa gtacactgag tcaataggga
     1201 ctttccattg ggttttgccc agtacaaaag gtcaataggg ggtgagtcaa tgggtttttc
     1261 ccattattgg cacatacata aggtcaatag gggtgactag tggagaagag catgcttgag
     1321 ggctgagtgc ccctcagtgg gcagagagca catggcccac agtccctgag aagttggggg
     1381 gaggggtggg caattgaact ggtgcctaga gaaggtgggg cttgggtaaa ctgggaaagt
     1441 gatgtggtgt actggctcca cctttttccc cagggtgggg gagaaccata tataagtgca
     1501 gtagtctctg tgaacattcg gtttagtgaa cctgcaggtc ggccgccacg accggtgcca
     1561 ccatgactga atacaaacca actgttcgcc tggcaactcg tgatgatgtt ccacgtgcag
     1621 ttcgcaccct ggctgctgca tttgctgact accctgcaac ccgtcacact gtggacccag
     1681 accgccacat tgaacgtgtg actgaactgc aggagctgtt cctgacccgt gtgggcctgg
     1741 acattggcaa agtgtgggtg gcagatgatg gtgctgctgt ggcagtgtgg accacccctg
     1801 aatctgttga agctggtgca gtgtttgctg agattggccc acgcatggca gaactgtctg
     1861 gcagccgcct ggcagcacaa cagcagatgg aaggtctgct ggcaccacac cgcccaaaag
     1921 aacctgcttg gttcctggca actgtgggtg tgagccctga ccaccagggt aagggcctgg
     1981 gctctgcagt ggtgctgcct ggtgtggaag cagctgaacg tgcaggtgtg cctgctttcc
     2041 tggagacctc agctccacgc aacctgcctt tctatgaacg cctgggcttc actgtgactg
     2101 ctgatgtgga agtgccagaa ggcccacgca cttggtgcat gactcgcaaa ccaggtgcta
     2161 gccttggctc cggcgagggc aggggaagtc ttctaacatg cggggacgtg gaggaaaatc
     2221 ccggcccact cggctacccc tacgacgtgc ccgactacgc cagcggcatg gtgagcaagg
     2281 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     2341 gccacaagtt cagcgtccgc ggcgagggcg agggcgatgc caccaacggc aagctgaccc
     2401 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct
     2461 tcggctacgg cgtggcctgc ttcagccgct accccgacca catgaagcag cacgacttct
     2521 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catctctttc aaggacgacg
     2581 gtacctacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     2641 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2701 acttcaacag ccacaacgtc tatatcacgg ccgacaagca gaagaacggc atcaaggcta
     2761 acttcaagat ccgccacaac gttgaggacg gcagcgtgca gctcgccgac cactaccagc
     2821 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagccatc
     2881 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     2941 tgaccgccgc cgggattaca catggcatgg acgagctgta caaggatatc catgcacact
     3001 ggcggccgct cgagatgaat cccagtatga agcagaaaca agaagaaatc aaagagaata
     3061 taaagaatag ttctgtccca agaagaactc tgaagatgat tcagccttct gcatctggat
     3121 ctcttgttgg aagagaaaat gagctgtccg caggcttgtc caaaaggaaa catcggaatg
     3181 accacttaac atctacaact tccagccctg gggttattgt cccagaatct agtgaaaata
     3241 aaaatcttgg aggagtcacc caggagtcat ttgatcttat gattaaagaa aatccatcct
     3301 ctcagtattg gaaggaagtg gcagaaaaac ggagaaaggc gctgtaagcc cctctccctc
     3361 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     3421 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     3481 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     3541 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     3601 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     3661 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     3721 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     3781 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
     3841 atgtgtttag tcgaggttaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     3901 ctttgaaaaa cacgatgata atggccacag aattcatggt gagcgtgatc aagcccgaga
     3961 tgaagatgag gtactacatg gacggctccg tcaatgggca tgagttcaca atcgagggtg
     4021 agggcacagg cagaccttac gagggacatc aggagatgac actgcgcgtc acaatggccg
     4081 agggcgggcc aatgcctttc gccttcgacc tggtgtccca cgtgttctgt tacggccaca
     4141 gagtttttac caagtaccca gaagagatcc cagactattt caagcaggcc tttcctgagg
     4201 gcctgtcctg ggagaggtcc ctggagttcg aggacggcgg ctccgcctcc gtgagcgccc
     4261 acatcagcct gaggggcaac accttctacc acaagtccaa gttcaccggc gtgaacttcc
     4321 ccgccgacgg ccccatcatg cagaaccaga gcgtggactg ggagccctcc accgagaaga
     4381 tcaccgccag cgacggcgtg ctgaagggcg acgtgaccat gtacctgaag ctggagggcg
     4441 gcggcaacca caagtgccag atgaagacca cctacaaggc cgccaaggag atcctggaga
     4501 tgcccggcga ccactacatc ggccacaggc tggtgaggaa gaccgagggc aacatcaccg
     4561 agcaggtgga ggacgccgtg gcccactact caatgttgcc ttccctgggc gatatccatg
     4621 cacactggcg gccgctcgag cccagccccg ccaggcccgc actccgcgcc ccggcctccg
     4681 ctaccagtgg cagccgcaag cgcgcccgcc cgcccgccgc ccccggacgc gaccaggcca
     4741 ggccaccggc ccgcaggaga ctgcggctgt cggtggacga ggtttccagc cccagtaccc
     4801 ccgaggcccc agacatccca gcctgccctt ctccgggcca gaagataaag aaatccaccc
     4861 cggcagcagg tcagccgccc cacctgacat ccgcgcagga ccaggacacc atctgagcta
     4921 gagttcgcgt ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     4981 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     5041 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     5101 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     5161 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     5221 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     5281 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     5341 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     5401 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     5461 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     5521 ccccgcctgg aattaattct ggctaagcac ttcgtggccg tcgatcgttt aaagggaggt
     5581 agtgagtcga gatatcgata ctagtgatcc agacatgata agatacattg atgagtttgg
     5641 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     5701 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     5761 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     5821 caaatgtggt atggctgatt atgatcctct agagggacag ccccccccca aagcccccag
     5881 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     5941 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     6001 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     6061 cacctggggg gatacgggga aaagccatac caatggagct cacggggaca gccccccccc
     6121 aaagccccca gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg
     6181 gggctccgct ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag
     6241 cccgggcacg gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt
     6301 gagcctgcag acacctgggg ggatacgggg aaaaagaatt cgataaaagt tttgttactt
     6361 tatagaagaa attttgagtt tttgtttttt ttaataaata aataaacata aataaattgt
     6421 ttgttgaatt tattattagt atgtaagtgt aaatataata aaacttaata tctattcaaa
     6481 ttaataaata aacctcgata tacagaccga taaaacacat gcgtcaattt tacacatgat
     6541 tatctttaac gtacgtcaca atatgattat ctttctaggg ttaatctagc tgcgtgttct
     6601 gcagcgtgtc gagcatcttc atctgctcca tcacgctgta aaacacattt gcaccgcgag
     6661 tctgcccgtc ctccacgggt tcaaaaacgt gaatgaacga ggcgcgctca ctggccgtcg
     6721 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     6781 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     6841 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     6901 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     6961 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     7021 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     7081 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     7141 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     7201 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     7261 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     7321 tttaggtggc 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 atgcctgtag 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 actcatatat
     8341 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     8401 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     8461 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     8521 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     8581 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     8641 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     8701 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     8761 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     8821 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     8881 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     8941 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     9001 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     9061 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     9121 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     9181 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     9241 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     9301 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     9361 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     9421 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     9481 ttacgccaag cgcgcccgcc gggtaactca cggggtatcc atgtccattt ctgcggcatc
     9541 cagccaggat acccgtcctc gctgacgtaa tatcccagcg ccgcaccgct gtcattaatc
     9601 tgcacaccgg cacggcagtt ccggctgtcg ccggtattgt tcgggttgct gatgcgcttc
     9661 gggctgacca tccggaactg tgtccggaaa agccgcgacg aactggtatc ccaggtggcc
     9721 tgaacgaaca gttcaccgtt aaaggcgtgc atggccacac cttcccgaat catcatggta
     9781 aacgtgcgtt ttcgctcaac gtcaatgcag cagcagtcat cctcggcaaa ctctttccat
     9841 gccgcttcaa cctcgcggga aaaggcacgg gcttcttcct ccccgatgcc cagatagcgc
     9901 cagcttgggc gatgactgag ccggaaaaaa gacccgacga tatgatcctg atgcagctag
     9961 a
//
