LOCUS       sequence-267621-        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"
     CDS             1263..1316
                     /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             1323..1349
                     /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             1323..1349
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(1356..1358,1359..1361,1362..2072)
                     /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(1401..1422)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2009..2030
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2103..2432
                     /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    2437..3009
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2604..2621)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2831..2850
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3024..3674
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAH"
     CDS             3024..3674
                     /codon_start=1
                     /product="humanized monomeric Kusabira-Orange fluorescent 
                     protein"
                     /label=hmKO
                     /note="mammalian codon-optimized"
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAH"
     CDS             3729..4001
                     /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    4026..4614
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4079..4099)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4497..4508)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4694..4828
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4718..4737)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4772..4791
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5607..5669
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5799..5816)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5799..5815)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5808..5830)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5957..6412
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6044..6063)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6254..6275
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6438..6542
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6543..6611,6612..7403)
                     /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(6761..6780)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7574..8162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8063..8082
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8316..8333
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8450..8471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8486..8491,8492..8509,8510..8516)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8524..8540
                     /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     8529..8551
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8548..8564
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8548..8564
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   9055..9089
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9358..9374)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9358..9374)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        9629..9877
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     9629..9649
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9800..9819
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 tcaataggga ctttccattg ggttttgccc agtacataag gtcaataggg ggtgagtcaa
       61 caggaaagtc ccattggagc caagtacatt gagtcaatag ggactttcca atgggttttg
      121 cccagtacat aaggtcaatg ggaggtaagc caatgggttt ttcccattac tgacatgtat
      181 actgagtcat tagggacttt ccaatgggtt ttgcccagta cataaggtca ataggggtga
      241 atcaacagga aagtcccatt ggagccaagt acactgagtc aatagggact ttccattggg
      301 ttttgcccag tacaaaaggt caataggggg tgagtcaatg ggtttttccc attattggca
      361 catacataag gtcaataggg gtgactagtg gagaagagca tgcttgaggg ctgagtgccc
      421 ctcagtgggc agagagcaca tggcccacag tccctgagaa gttgggggga ggggtgggca
      481 attgaactgg tgcctagaga aggtggggct tgggtaaact gggaaagtga tgtggtgtac
      541 tggctccacc tttttcccca gggtggggga gaaccatata taagtgcagt agtctctgtg
      601 aacattcggt ttagtgaacc tgcaggtcgg ccgccacgac cggtgccacc atgactgaat
      661 acaaaccaac tgttcgcctg gcaactcgtg atgatgttcc acgtgcagtt cgcaccctgg
      721 ctgctgcatt tgctgactac cctgcaaccc gtcacactgt ggacccagac cgccacattg
      781 aacgtgtgac tgaactgcag gagctgttcc tgacccgtgt gggcctggac attggcaaag
      841 tgtgggtggc agatgatggt gctgctgtgg cagtgtggac cacccctgaa tctgttgaag
      901 ctggtgcagt gtttgctgag attggcccac gcatggcaga actgtctggc agccgcctgg
      961 cagcacaaca gcagatggaa ggtctgctgg caccacaccg cccaaaagaa cctgcttggt
     1021 tcctggcaac tgtgggtgtg agccctgacc accagggtaa gggcctgggc tctgcagtgg
     1081 tgctgcctgg tgtggaagca gctgaacgtg caggtgtgcc tgctttcctg gagacctcag
     1141 ctccacgcaa cctgcctttc tatgaacgcc tgggcttcac tgtgactgct gatgtggaag
     1201 tgccagaagg cccacgcact tggtgcatga ctcgcaaacc aggtgctagc cttggctccg
     1261 gcgagggcag gggaagtctt ctaacatgcg gggacgtgga ggaaaatccc ggcccactcg
     1321 gctaccccta cgacgtgccc gactacgcca gcggcatggt gagcaagggc gaggagctgt
     1381 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     1441 gcgtccgcgg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatct
     1501 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcg
     1561 tggcctgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     1621 tgcccgaagg ctacgtccag gagcgcacca tctctttcaa ggacgacggt acctacaaga
     1681 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     1741 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac ttcaacagcc
     1801 acaacgtcta tatcacggcc gacaagcaga agaacggcat caaggctaac ttcaagatcc
     1861 gccacaacgt tgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     1921 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagccatcag tccgccctga
     1981 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     2041 ggattacaca tggcatggac gagctgtaca aggatatcca tgcacactgg cggccgctcg
     2101 agatgaatcc cagtatgaag cagaaacaag aagaaatcaa agagaatata aagaatagtt
     2161 ctgtcccaag aagaactctg aagatgattc agccttctgc atctggatct cttgttggaa
     2221 gagaaaatga gctgtccgca ggcttgtcca aaaggaaaca tcggaatgac cacttaacat
     2281 ctacaacttc cagccctggg gttattgtcc cagaatctag tgaaaataaa aatcttggag
     2341 gagtcaccca ggagtcattt gatcttatga ttaaagaaaa tccatcctct cagtattgga
     2401 aggaagtggc agaaaaacgg agaaaggcgc tgtaagcccc tctccctccc ccccccctaa
     2461 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     2521 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     2581 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     2641 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     2701 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     2761 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     2821 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     2881 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
     2941 gaggttaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     3001 cgatgataat ggccacagaa ttcatggtga gcgtgatcaa gcccgagatg aagatgaggt
     3061 actacatgga cggctccgtc aatgggcatg agttcacaat cgagggtgag ggcacaggca
     3121 gaccttacga gggacatcag gagatgacac tgcgcgtcac aatggccgag ggcgggccaa
     3181 tgcctttcgc cttcgacctg gtgtcccacg tgttctgtta cggccacaga gtttttacca
     3241 agtacccaga agagatccca gactatttca agcaggcctt tcctgagggc ctgtcctggg
     3301 agaggtccct ggagttcgag gacggcggct ccgcctccgt gagcgcccac atcagcctga
     3361 ggggcaacac cttctaccac aagtccaagt tcaccggcgt gaacttcccc gccgacggcc
     3421 ccatcatgca gaaccagagc gtggactggg agccctccac cgagaagatc accgccagcg
     3481 acggcgtgct gaagggcgac gtgaccatgt acctgaagct ggagggcggc ggcaaccaca
     3541 agtgccagat gaagaccacc tacaaggccg ccaaggagat cctggagatg cccggcgacc
     3601 actacatcgg ccacaggctg gtgaggaaga ccgagggcaa catcaccgag caggtggagg
     3661 acgccgtggc ccactactca atgttgcctt ccctgggcga tatccatgca cactggcggc
     3721 cgctcgagcc cagccccgcc aggcccgcac tccgcgcccc ggcctccgct accagtggca
     3781 gccgcaagcg cgcccgcccg cccgccgccc ccggacgcga ccaggccagg ccaccggccc
     3841 gcaggagact gcggctgtcg gtggacgagg tttccagccc cagtaccccc gaggccccag
     3901 acatcccagc ctgcccttct ccgggccaga agataaagaa atccaccccg gcagcaggtc
     3961 agccgcccca cctgacatcc gcgcaggacc aggacaccat ctgagctaga gttcgcgtct
     4021 ggaacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     4081 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     4141 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     4201 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     4261 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     4321 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     4381 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     4441 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     4501 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     4561 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     4621 ttaattctgg ctaagcactt cgtggccgtc gatcgtttaa agggaggtag tgagtcgaga
     4681 tatcgatact agtgatccag acatgataag atacattgat gagtttggac aaaccacaac
     4741 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     4801 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     4861 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     4921 ggctgattat gatcctctag agggacagcc cccccccaaa gcccccaggg atgtaattac
     4981 gtccctcccc cgctaggggg cagcagcgag ccgcccgggg ctccgctccg gtccggcgct
     5041 ccccccgcat ccccgagccg gcagcgtgcg gggacagccc gggcacgggg aaggtggcac
     5101 gggatcgctt tcctctgaac gcttctcgct gctctttgag cctgcagaca cctgggggga
     5161 tacggggaaa aagccatacc aatggagctc acggggacag ccccccccca aagcccccag
     5221 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     5281 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     5341 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     5401 cacctggggg gatacgggga aaaagaattc gataaaagtt ttgttacttt atagaagaaa
     5461 ttttgagttt ttgttttttt taataaataa ataaacataa ataaattgtt tgttgaattt
     5521 attattagta tgtaagtgta aatataataa aacttaatat ctattcaaat taataaataa
     5581 acctcgatat acagaccgat aaaacacatg cgtcaatttt acacatgatt atctttaacg
     5641 tacgtcacaa tatgattatc tttctagggt taatctagct gcgtgttctg cagcgtgtcg
     5701 agcatcttca tctgctccat cacgctgtaa aacacatttg caccgcgagt ctgcccgtcc
     5761 tccacgggtt caaaaacgtg aatgaacgag gcgcgctcac tggccgtcgt tttacaacgt
     5821 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     5881 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     5941 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     6001 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     6061 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     6121 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     6181 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     6241 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     6301 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     6361 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     6421 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     6481 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6541 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6601 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6661 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6721 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6781 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6841 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6901 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6961 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     7021 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     7081 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     7141 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     7201 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     7261 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     7321 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     7381 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     7441 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     7501 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     7561 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     7621 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     7681 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     7741 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     7801 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     7861 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7921 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7981 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     8041 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     8101 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     8161 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     8221 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     8281 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     8341 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     8401 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     8461 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     8521 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     8581 gcgcccgccg ggtaactcac ggggtatcca tgtccatttc tgcggcatcc agccaggata
     8641 cccgtcctcg ctgacgtaat atcccagcgc cgcaccgctg tcattaatct gcacaccggc
     8701 acggcagttc cggctgtcgc cggtattgtt cgggttgctg atgcgcttcg ggctgaccat
     8761 ccggaactgt gtccggaaaa gccgcgacga actggtatcc caggtggcct gaacgaacag
     8821 ttcaccgtta aaggcgtgca tggccacacc ttcccgaatc atcatggtaa acgtgcgttt
     8881 tcgctcaacg tcaatgcagc agcagtcatc ctcggcaaac tctttccatg ccgcttcaac
     8941 ctcgcgggaa aaggcacggg cttcttcctc cccgatgccc agatagcgcc agcttgggcg
     9001 atgactgagc cggaaaaaag acccgacgat atgatcctga tgcagctaga ttaaccctag
     9061 aaagatagtc tgcgtaaaat tgacgcatgc attcttgaaa tattgctctc tctttctaaa
     9121 tagcgcgaat ccgtcgctgt gcatttagga catctcagtc gccgcttgga gctcccgtga
     9181 ggcgtgcttg tcaatgcggt aagtgtcact gattttgaac tataacgacc gcgtgagtca
     9241 aaatgacgca tgattatctt ttacgtgact tttaagattt aactcatacg ataattatat
     9301 tgttatttca tgttctactt acgtgataac ttattatata tatattttct tgttatagat
     9361 ccactagttc tagagggaca gccccccccc aaagccccca gggatgtaat tacgtccctc
     9421 ccccgctagg gggcagcagc gagccgcccg gggctccgct ccggtccggc gctccccccg
     9481 catccccgag ccggcagcgt gcggggacag cccgggcacg gggaaggtgg cacgggatcg
     9541 ctttcctctg aacgcttctc gctgctcttt gagcctgcag acacctgggg ggatacgggg
     9601 aaaaggcctc cagggccgcc ccttcaccga gggcctattt cccatgattc cttcatattt
     9661 gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa
     9721 gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt
     9781 aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt
     9841 cttggcttta tatatcttgt ggaaaggacg aaacaccggg agacgctcga cccgtctcgt
     9901 ttttgcctcc aaggcctgca ggagtcaatg ggaaaaaccc attggagcca agtacactga
     9961 c
//
