LOCUS       sequence-267623-        9989 bp    DNA     circular SYN 15-APR-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 9989)
  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 9989)
  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: "Am J 
            Physiol Gastrointest Liver Physiol. 2023 Jun 13. doi: 
            10.1152/ajpgi.00058.2022."
COMMENT     Sequence Label: BII-Sh-FUCCI-Luc
FEATURES             Location/Qualifiers
     source          1..9989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(269..285)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(269..285)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        540..788
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     540..560
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     711..730
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     CDS             2163..2216
                     /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             2223..2249
                     /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             2223..2249
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(2256..2258,2259..2261,2262..2972)
                     /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(2301..2322)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2909..2930
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3003..3332
                     /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    3337..3909
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3504..3521)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3731..3750
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3924..4574
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAH"
     CDS             3924..4574
                     /codon_start=1
                     /product="humanized monomeric Kusabira-Orange fluorescent 
                     protein"
                     /label=hmKO
                     /note="mammalian codon-optimized"
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAH"
     CDS             4629..4901
                     /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    4926..5514
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4979..4999)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5397..5408)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5594..5728
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5618..5637)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5672..5691
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   6507..6569
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6699..6716)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6699..6715)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6708..6730)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6857..7312
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6944..6963)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7154..7175
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7338..7442
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7443..7511,7512..8303)
                     /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(7661..7680)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8474..9062
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8963..8982
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9216..9233
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9350..9371
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9386..9391,9392..9409,9410..9416)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9424..9440
                     /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     9429..9451
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9448..9464
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9448..9464
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   9955..9989
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 cattcttgaa atattgctct ctctttctaa atagcgcgaa tccgtcgctg tgcatttagg
       61 acatctcagt cgccgcttgg agctcccgtg aggcgtgctt gtcaatgcgg taagtgtcac
      121 tgattttgaa ctataacgac cgcgtgagtc aaaatgacgc atgattatct tttacgtgac
      181 ttttaagatt taactcatac gataattata ttgttatttc atgttctact tacgtgataa
      241 cttattatat atatattttc ttgttataga tccactagtt ctagagggac agcccccccc
      301 caaagccccc agggatgtaa ttacgtccct cccccgctag ggggcagcag cgagccgccc
      361 ggggctccgc tccggtccgg cgctcccccc gcatccccga gccggcagcg tgcggggaca
      421 gcccgggcac ggggaaggtg gcacgggatc gctttcctct gaacgcttct cgctgctctt
      481 tgagcctgca gacacctggg gggatacggg gaaaaggcct ccagggccgc cccttcaccg
      541 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
      601 taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
      661 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata
      721 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac
      781 gaaacaccgg tcacagaatc gtcgtatgca gctcgagctg catacgacga ttctgtgatt
      841 tttgcctcca aggcctgcag gagtcaatgg gaaaaaccca ttggagccaa gtacactgac
      901 tcaataggga ctttccattg ggttttgccc agtacataag gtcaataggg ggtgagtcaa
      961 caggaaagtc ccattggagc caagtacatt gagtcaatag ggactttcca atgggttttg
     1021 cccagtacat aaggtcaatg ggaggtaagc caatgggttt ttcccattac tgacatgtat
     1081 actgagtcat tagggacttt ccaatgggtt ttgcccagta cataaggtca ataggggtga
     1141 atcaacagga aagtcccatt ggagccaagt acactgagtc aatagggact ttccattggg
     1201 ttttgcccag tacaaaaggt caataggggg tgagtcaatg ggtttttccc attattggca
     1261 catacataag gtcaataggg gtgactagtg gagaagagca tgcttgaggg ctgagtgccc
     1321 ctcagtgggc agagagcaca tggcccacag tccctgagaa gttgggggga ggggtgggca
     1381 attgaactgg tgcctagaga aggtggggct tgggtaaact gggaaagtga tgtggtgtac
     1441 tggctccacc tttttcccca gggtggggga gaaccatata taagtgcagt agtctctgtg
     1501 aacattcggt ttagtgaacc tgcaggtcgg ccgccacgac cggtgccacc atgactgaat
     1561 acaaaccaac tgttcgcctg gcaactcgtg atgatgttcc acgtgcagtt cgcaccctgg
     1621 ctgctgcatt tgctgactac cctgcaaccc gtcacactgt ggacccagac cgccacattg
     1681 aacgtgtgac tgaactgcag gagctgttcc tgacccgtgt gggcctggac attggcaaag
     1741 tgtgggtggc agatgatggt gctgctgtgg cagtgtggac cacccctgaa tctgttgaag
     1801 ctggtgcagt gtttgctgag attggcccac gcatggcaga actgtctggc agccgcctgg
     1861 cagcacaaca gcagatggaa ggtctgctgg caccacaccg cccaaaagaa cctgcttggt
     1921 tcctggcaac tgtgggtgtg agccctgacc accagggtaa gggcctgggc tctgcagtgg
     1981 tgctgcctgg tgtggaagca gctgaacgtg caggtgtgcc tgctttcctg gagacctcag
     2041 ctccacgcaa cctgcctttc tatgaacgcc tgggcttcac tgtgactgct gatgtggaag
     2101 tgccagaagg cccacgcact tggtgcatga ctcgcaaacc aggtgctagc cttggctccg
     2161 gcgagggcag gggaagtctt ctaacatgcg gggacgtgga ggaaaatccc ggcccactcg
     2221 gctaccccta cgacgtgccc gactacgcca gcggcatggt gagcaagggc gaggagctgt
     2281 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     2341 gcgtccgcgg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatct
     2401 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcg
     2461 tggcctgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2521 tgcccgaagg ctacgtccag gagcgcacca tctctttcaa ggacgacggt acctacaaga
     2581 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     2641 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac ttcaacagcc
     2701 acaacgtcta tatcacggcc gacaagcaga agaacggcat caaggctaac ttcaagatcc
     2761 gccacaacgt tgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     2821 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagccatcag tccgccctga
     2881 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     2941 ggattacaca tggcatggac gagctgtaca aggatatcca tgcacactgg cggccgctcg
     3001 agatgaatcc cagtatgaag cagaaacaag aagaaatcaa agagaatata aagaatagtt
     3061 ctgtcccaag aagaactctg aagatgattc agccttctgc atctggatct cttgttggaa
     3121 gagaaaatga gctgtccgca ggcttgtcca aaaggaaaca tcggaatgac cacttaacat
     3181 ctacaacttc cagccctggg gttattgtcc cagaatctag tgaaaataaa aatcttggag
     3241 gagtcaccca ggagtcattt gatcttatga ttaaagaaaa tccatcctct cagtattgga
     3301 aggaagtggc agaaaaacgg agaaaggcgc tgtaagcccc tctccctccc ccccccctaa
     3361 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     3421 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     3481 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     3541 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     3601 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     3661 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     3721 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     3781 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
     3841 gaggttaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     3901 cgatgataat ggccacagaa ttcatggtga gcgtgatcaa gcccgagatg aagatgaggt
     3961 actacatgga cggctccgtc aatgggcatg agttcacaat cgagggtgag ggcacaggca
     4021 gaccttacga gggacatcag gagatgacac tgcgcgtcac aatggccgag ggcgggccaa
     4081 tgcctttcgc cttcgacctg gtgtcccacg tgttctgtta cggccacaga gtttttacca
     4141 agtacccaga agagatccca gactatttca agcaggcctt tcctgagggc ctgtcctggg
     4201 agaggtccct ggagttcgag gacggcggct ccgcctccgt gagcgcccac atcagcctga
     4261 ggggcaacac cttctaccac aagtccaagt tcaccggcgt gaacttcccc gccgacggcc
     4321 ccatcatgca gaaccagagc gtggactggg agccctccac cgagaagatc accgccagcg
     4381 acggcgtgct gaagggcgac gtgaccatgt acctgaagct ggagggcggc ggcaaccaca
     4441 agtgccagat gaagaccacc tacaaggccg ccaaggagat cctggagatg cccggcgacc
     4501 actacatcgg ccacaggctg gtgaggaaga ccgagggcaa catcaccgag caggtggagg
     4561 acgccgtggc ccactactca atgttgcctt ccctgggcga tatccatgca cactggcggc
     4621 cgctcgagcc cagccccgcc aggcccgcac tccgcgcccc ggcctccgct accagtggca
     4681 gccgcaagcg cgcccgcccg cccgccgccc ccggacgcga ccaggccagg ccaccggccc
     4741 gcaggagact gcggctgtcg gtggacgagg tttccagccc cagtaccccc gaggccccag
     4801 acatcccagc ctgcccttct ccgggccaga agataaagaa atccaccccg gcagcaggtc
     4861 agccgcccca cctgacatcc gcgcaggacc aggacaccat ctgagctaga gttcgcgtct
     4921 ggaacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     4981 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     5041 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     5101 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     5161 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     5221 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     5281 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     5341 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     5401 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     5461 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     5521 ttaattctgg ctaagcactt cgtggccgtc gatcgtttaa agggaggtag tgagtcgaga
     5581 tatcgatact agtgatccag acatgataag atacattgat gagtttggac aaaccacaac
     5641 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     5701 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     5761 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     5821 ggctgattat gatcctctag agggacagcc cccccccaaa gcccccaggg atgtaattac
     5881 gtccctcccc cgctaggggg cagcagcgag ccgcccgggg ctccgctccg gtccggcgct
     5941 ccccccgcat ccccgagccg gcagcgtgcg gggacagccc gggcacgggg aaggtggcac
     6001 gggatcgctt tcctctgaac gcttctcgct gctctttgag cctgcagaca cctgggggga
     6061 tacggggaaa aagccatacc aatggagctc acggggacag ccccccccca aagcccccag
     6121 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     6181 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     6241 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     6301 cacctggggg gatacgggga aaaagaattc gataaaagtt ttgttacttt atagaagaaa
     6361 ttttgagttt ttgttttttt taataaataa ataaacataa ataaattgtt tgttgaattt
     6421 attattagta tgtaagtgta aatataataa aacttaatat ctattcaaat taataaataa
     6481 acctcgatat acagaccgat aaaacacatg cgtcaatttt acacatgatt atctttaacg
     6541 tacgtcacaa tatgattatc tttctagggt taatctagct gcgtgttctg cagcgtgtcg
     6601 agcatcttca tctgctccat cacgctgtaa aacacatttg caccgcgagt ctgcccgtcc
     6661 tccacgggtt caaaaacgtg aatgaacgag gcgcgctcac tggccgtcgt tttacaacgt
     6721 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     6781 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     6841 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     6901 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     6961 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     7021 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     7081 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     7141 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     7201 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     7261 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     7321 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     7381 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     7441 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     7501 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     7561 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     7621 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     7681 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     7741 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     7801 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     7861 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     7921 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     7981 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     8041 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     8101 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     8161 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     8221 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     8281 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     8341 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     8401 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     8461 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     8521 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     8581 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     8641 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     8701 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     8761 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     8821 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     8881 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     8941 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     9001 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     9061 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     9121 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     9181 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     9241 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     9301 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     9361 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     9421 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     9481 gcgcccgccg ggtaactcac ggggtatcca tgtccatttc tgcggcatcc agccaggata
     9541 cccgtcctcg ctgacgtaat atcccagcgc cgcaccgctg tcattaatct gcacaccggc
     9601 acggcagttc cggctgtcgc cggtattgtt cgggttgctg atgcgcttcg ggctgaccat
     9661 ccggaactgt gtccggaaaa gccgcgacga actggtatcc caggtggcct gaacgaacag
     9721 ttcaccgtta aaggcgtgca tggccacacc ttcccgaatc atcatggtaa acgtgcgttt
     9781 tcgctcaacg tcaatgcagc agcagtcatc ctcggcaaac tctttccatg ccgcttcaac
     9841 ctcgcgggaa aaggcacggg cttcttcctc cccgatgccc agatagcgcc agcttgggcg
     9901 atgactgagc cggaaaaaag acccgacgat atgatcctga tgcagctaga ttaaccctag
     9961 aaagatagtc tgcgtaaaat tgacgcatg
//
