LOCUS       sequence-334275-       15193 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible ZEB2 expression from AAVS1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15193)
  AUTHORS   Benito-Kwiecinski S, Giandomenico SL, Sutcliffe M, Riis ES, 
            Freire-Pritchett P, Kelava I, Wunderlich S, Martin U, Wray GA, 
            McDole K, Lancaster MA
  TITLE     An early cell shape transition drives evolutionary expansion of the 
            human forebrain.
  JOURNAL   Cell. 2021 Apr 15;184(8):2084-2102.e19. doi: 
            10.1016/j.cell.2021.02.050. Epub 2021 Mar 24.
  PUBMED    33765444
REFERENCE   2  (bases 1 to 15193)
  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: 
            "10.1016/j.cell.2021.02.050"; journalName: "Cell"; date: 
            "2021-04-15- 15"; volume: "184"; issue: "8"; pages: "2084-2102.e19"
COMMENT     Sequence Label: AAVS1-Puro-CAG-fl-STOP-fl-ZEB2-GFP-Flag-WPRE-p...
FEATURES             Location/Qualifiers
     source          1..15193
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      23..611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     512..531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     765..782
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    899..920
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(935..940,941..958,959..965)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    973..989
                     /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     978..1000
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     997..1013
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     997..1013
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1034..1052
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    1087..1890
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     misc_feature    1897..1922
                     /label=SA
                     /note="splice acceptor site"
     CDS             1946..1999
                     /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             2009..2608
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2009..2028)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2505..2525
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2640..2657)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2646..2870
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        3230..3533
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3535..3791
                     /label=chicken beta-actin promoter
                     /note="contains a 21-bp deletion relative to the standard 
                     chicken beta-actin promoter"
     intron          3792..4808
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4732..4754
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4816..4835
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    4903..4936
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    5103..5237
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    5357..5491
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    5611..5745
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    5774..5807
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             join(9606..9608,9609..9611,9612..10322)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(9606..9608,9609..9611,9612..10322)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(9651..9672)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(9912..9931)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     10259..10280
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             10338..10361
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    10410..10998
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10463..10483)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10881..10892)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    11154..11209
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     polyA_signal    11392..11447
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(11446..11465)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     misc_feature    11483..12319
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(12353..12372)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(12354..12372)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(12379..12396)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(12379..12395)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12388..12410)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             12533..12835
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     12714..12733
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(13238..13257)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     13852..13871
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(14232..15023,15024..15092))
                     /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     14855..14874
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
       61 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
      121 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
      181 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
      241 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
      301 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
      361 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
      421 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
      481 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
      541 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      601 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      661 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
      721 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
      781 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
      841 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
      901 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
      961 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     1021 acgccaagct cagaattaac cctcactaaa gggactagtc ctgcaggttt aaacgaattc
     1081 gccctttgct ttctctgacc agcattctct cccctgggcc tgtgccgctt tctgtctgca
     1141 gcttgtggcc tgggtcacct ctacggctgg cccagatcct tccctgccgc ctccttcagg
     1201 ttccgtcttc ctccactccc tcttcccctt gctctctgct gtgttgctgc ccaaggatgc
     1261 tctttccgga gcacttcctt ctcggcgctg caccacgtga tgtcctctga gcggatcctc
     1321 cccgtgtctg ggtcctctcc gggcatctct cctccctcac ccaaccccat gccgtcttca
     1381 ctcgctgggt tcccttttcc ttctccttct ggggcctgtg ccatctctcg tttcttagga
     1441 tggccttctc cgacggatgt ctcccttgcg tcccgcctcc ccttcttgta ggcctgcatc
     1501 atcaccgttt ttctggacaa ccccaaagta ccccgtctcc ctggctttag ccacctctcc
     1561 atcctcttgc tttctttgcc tggacacccc gttctcctgt ggattcgggt cacctctcac
     1621 tcctttcatt tgggcagctc ccctaccccc cttacctctc tagtctgtgc tagctcttcc
     1681 agccccctgt catggcatct tccaggggtc cgagagctca gctagtcttc ttcctccaac
     1741 ccgggcccct atgtccactt caggacagca tgtttgctgc ctccagggat cctgtgtccc
     1801 cgagctggga ccaccttata ttcccagggc cggttaatgt ggctctggtt ctgggtactt
     1861 ttatctgtcc cctccacccc acagtggggc aagcttctga cctcttctct tcctcccaca
     1921 gggcctcgag agatctggca gcggagaggg cagaggaagt cttctaacat gcggtgacgt
     1981 ggaggagaat cccggcccta ggctcgagat gaccgagtac aagcccacgg tgcgcctcgc
     2041 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     2101 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     2161 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     2221 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     2281 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     2341 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     2401 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     2461 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     2521 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     2581 gtgcatgacc cgcaagcccg gtgcctgatc tagagggccc gtttaaaccc gctgatcagc
     2641 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2701 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2761 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2821 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg gtcgagatct
     2881 agttttaagt tggagtcatt aaaactcgtt tttcaactac tccacaaatt tcttgttaac
     2941 aaacaatagt tttggcaagt cagttaggac atctactttg tgcatgacac aagtcatttt
     3001 tccaacaatt gtttacagac agattatttc acttataatt cactgtatca caattccagt
     3061 gggtcagaag tttacataca ctaagttgac tgtgccttta aacagcttgg aaaattccag
     3121 aaaatgatgt catggcttta gaagcttcct gacctggtgc gcattagtct agttattaat
     3181 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     3241 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     3301 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     3361 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     3421 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     3481 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg
     3541 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt
     3601 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg gggggggggg
     3661 gcgrggggsg gggsggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc
     3721 tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg
     3781 cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct
     3841 cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg
     3901 gcccttctcc tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt
     3961 ggctgcgtga aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg
     4021 ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg
     4081 gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga
     4141 gcgcggccgg gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg
     4201 tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac
     4261 cccccctgca cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc
     4321 gtacggggcg tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc
     4381 cgggcggggc ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc
     4441 ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt
     4501 gcgagagggc gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc
     4561 gccgccgcac cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa
     4621 tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc
     4681 ggggctgtcc gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct
     4741 tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt
     4801 tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat
     4861 tcgaaagtat aggaacttcg tcgacattta aatcatttaa atataacttc gtataatgta
     4921 tgctatacga agttattcgc gatgaataaa tgaaagcttg cagatctgcg actctagagg
     4981 atctgcgact ctagaggatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     5041 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     5101 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5161 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5221 cttatcatgt ctggatctgc gactctagag gatcataatc agccatacca catttgtaga
     5281 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     5341 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5401 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5461 actcatcaat gtatcttatc atgtctggat ctgcgactct agaggatcat aatcagccat
     5521 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     5581 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     5641 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5701 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccccat caagctgatc
     5761 cggaaccctt aatataactt cgtataatgt atgctatacg aagttattag gtccctcgac
     5821 cggcgcgaac tggatccact agtccagtgt ggtggaattc tgcagatacg cgtatgaagc
     5881 agccgatcat ggcggatggc ccccggtgca agaggcgcaa acaagccaat cccaggagga
     5941 aaaacgtggt gaactatgac aatgtagtgg acacaggttc tgaaacagat gaggaagaca
     6001 agcttcatat tgctgaggat gacggtattg ccaaccctct ggaccaggag acgagtccag
     6061 ctagtgtgcc caaccatgag tcctccccac acgtgagcca agctctgttg ccaagagagg
     6121 aagaggaaga tgaaataagg gagggtggag tggaacaccc ctggcacaac aacgagattc
     6181 tacaagcctc tgtagatggt ccagaagaaa tgaaggaaga ctatgacact atggggccag
     6241 aagccacgat ccagaccgca attaacaatg gtacagtgaa gaatgcaaat tgcacatcag
     6301 attttgagga atactttgcc aaaagaaaac tggaggaacg cgatggtcat gcagtcagca
     6361 tcgaggagta ccttcagcgc agtgacacag ccattattta cccagaagcc cctgaggagc
     6421 tgtctcgcct tggcacgcca gaggccaatg ggcaagaaga aaatgacctg ccacctggaa
     6481 ctccagatgc ttttgcccaa ctgctgacct gcccctactg cgaccggggc tacaagcgct
     6541 tgacatcact gaaggagcac atcaagtacc gccacgagaa gaatgaagag aacttttcct
     6601 gccctctctg tagctacacg tttgcctacc gcacccagct cgagcggcat atggtgacac
     6661 acaagccagg gacagatcag caccaaatgc taacccaagg agcaggtaat cgcaagttca
     6721 aatgcacaga gtgtggcaag gccttcaaat ataaacacca tctgaaagaa cacctgcgaa
     6781 ttcacagtgg tgaaaaacct tacgagtgcc caaactgcaa gaaacgtttc tcccattctg
     6841 gttcctacag ttcgcacatc agcagcaaga aatgtattgg tttaatctct gtaaatggcc
     6901 gaatgagaaa caatatcaag acgggttctt cccctaattc tgtttcttct tctcctacta
     6961 attcagccat tacccagtta agaaacaagt tggagaatgg aaaaccactt agtatgtctg
     7021 aacagacagg cttacttaaa attaaaacag aaccactaga cttcaatgac tataaagttc
     7081 ttatggctac acacgggttt agtggcacta gtccctttat gaatggtggg cttggagcca
     7141 ccagcccttt aggagttcat ccatctgctc agagtccaat gcagcactta ggtgtaggga
     7201 tggaagcccc tttacttggg tttcccacca tgaatagtaa tttaagtgag gtacaaaagg
     7261 ttctacagat tgtggacaat actgtttcca ggcaaaaaat ggactgcaag gctgaagaaa
     7321 tttcaaagtt gaaaggttat cacatgaagg atccatgctc tcaacctgag gaacaaggag
     7381 ttacttctcc taatattccg cctgtcggtc ttccggtagt gagtcataat ggtgccacta
     7441 aaagtattat tgactatacg ttggaaaaag tcaatgaagc caaagcttgc ctccagagct
     7501 tgactactga ctcaaggaga cagatcagta atataaagaa agagaagcta cgtactttaa
     7561 tagatttggt cactgatgac aaaatgattg agaaccacaa catatccact ccattttcat
     7621 gccagttctg taaagaaagt tttcctggcc ccatcccttt gcatcagcat gaacgttacc
     7681 tttgtaagat gaatgaagag atcaaggcgg tcctgcagcc tcatgaaaac atagtcccca
     7741 acaaagccgg agtttttgtt gataataaag ccctcctctt gtcatctgta ctttctgaga
     7801 aaggaatgac aagccccatc aacccataca aggaccacat gtctgtactc aaagcatact
     7861 atgctatgaa catggagccc aactccgatg aactgctgaa aatttccatt gctgtgggcc
     7921 ttcctcagga atttgtgaag gaatggtttg aacaacgaaa agtctaccag tactcaaatt
     7981 ccaggtcccc atccctggaa agaagctcca agccgttagc tcccaacagt aaccctccca
     8041 caaaagactc tttattaccc aggtctcctg taaaacctat ggactccata acatcaccat
     8101 ctatagcaga actccacaac agtgttacga attgtgatcc tcctctcagg ctaacaaaac
     8161 cttcccattt taccaatatt aaaccagttg aaaaattgga ccactccagg agtaatactc
     8221 cttctccctt aaatctttcc tccacatctt ctaaaaactc ccacagtagt tcatacactc
     8281 caaacagctt ctcttctgag gagctccagg ctgagccttt agacttgtca ttaccaaaac
     8341 aaatgaaaga acccaaaagt attatagcca caaagaacaa aacaaaagct agtagcatca
     8401 gtttagatca taacagtgtt tcttcctcat ctgaaaactc agatgagcct ctgaacttga
     8461 cttttatcaa gaaggaattt tcaaattcaa ataatctgga caacaaaagc actaacccag
     8521 tgttcagcat gaacccattt agtgccaaac ctttatacac agctcttcca cctcaaagcg
     8581 catttccccc tgctactttc atgccaccag tccagaccag tattcctggg ctacgaccat
     8641 acccaggact ggatcagatg agcttcctac cacatatggc ctacacctac ccaactggag
     8701 cagctacttt tgctgatatg cagcaaagga gaaagtacca gcggaaacaa ggatttcagg
     8761 gagaattgct tgatggagca caagactaca tgtcaggcct agatgatatg acagactccg
     8821 actcctgtct gtctcgcaaa aagatcaaga agacagagag tggcatgtat gcatgtgact
     8881 tatgtgacaa gacattccag aaaagcagtt cccttctgcg acataaatac gaacacacag
     8941 gaaaaagacc acatcagtgt cagatttgta agaaagcgtt taaacacaag caccacctta
     9001 tcgagcactc aaggcttcac tcgggcgaga agccctatca gtgtgataaa tgtggcaagc
     9061 gcttctcaca ctcgggctcg tactcgcagc acatgaatca caggtattcc tactgcaagc
     9121 gggaggcgga ggagcgggaa gcggcggagc gcgaggcgcg cgagaaaggg cacttggaac
     9181 ccaccgagct gctgatgaac cgggcttact tgcagagcat tacccctcag gggtactctg
     9241 actcggagga gagggagagt atgccgaggg atggcgagag cgagaaggag cacgagaaag
     9301 aaggcgagga tggctacggg aagctgggca gacaggatgg cgacgaggag ttcgaggagg
     9361 aagaggaaga aagtgaaaat aaaagtatgg atacggatcc cgaaacgata cgagatgaag
     9421 aagagactgg agatcactcc atggacgata gttcggagga tgggaaaatg gaaaccaaat
     9481 cagaccacga ggaagacaat atggaagatg gcatgggtgg cggtggatcc ggtggcggtg
     9541 gatccggtgg cggtggatcc ggtggcggtg gatccggtgg cggtggatcc ggtggcggtg
     9601 gatccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     9661 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     9721 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     9781 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     9841 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     9901 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     9961 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
    10021 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
    10081 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
    10141 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
    10201 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
    10261 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
    10321 agggtggcgg tggatccgat tacaaggatg acgacgataa gtaaggtacc gcggccgcct
    10381 cgagaagctt cggggactag tcatatgata atcaacctct ggattacaaa atttgtgaaa
    10441 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
    10501 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
    10561 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
    10621 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
    10681 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
    10741 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
    10801 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
    10861 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
    10921 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
    10981 tttgggccgc ctccccgcat cggtacaatt cactcctcag gtgcaggctg cctatcagaa
    11041 ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc
    11101 tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag
    11161 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggggccgg
    11221 ccaggcaaaa aagaaaaagt aatgaattca ctcctcaggt gcaggctgcc tatcagaagg
    11281 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
    11341 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
    11401 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
    11461 atgggagggc aaatggtacc ttactaggga caggattggt gacagaaaag ccccatcctt
    11521 aggcctcctc cttcctagtc tcctgatatt gggtctaacc cccacctcct gttaggcaga
    11581 ttccttatct ggtgacacac ccccatttcc tggagccatc tctctccttg ccagaacctc
    11641 taaggtttgc ttacgatgga gccagagagg atcctgggag ggagagcttg gcagggggtg
    11701 ggagggaagg gggggatgcg tgacctgccc ggttctcagt ggccaccctg cgctaccctc
    11761 tcccagaacc tgagctgctc tgacgcggct gtctggtgcg tttcactgat cctggtgctg
    11821 cagcttcctt acacttccca agaggagaag cagtttggaa aaacaaaatc agaataagtt
    11881 ggtcctgagt tctaactttg gctcttcacc tttctagtcc ccaatttata ttgttcctcc
    11941 gtgcgtcagt tttacctgtg agataaggcc agtagccagc cccgtcctgg cagggctgtg
    12001 gtgaggaggg gggtgtccgt gtggaaaact ccctttgtga gaatggtgcg tcctaggtgt
    12061 tcaccaggtc gtggccgcct ctactccctt tctctttctc catccttctt tccttaaaga
    12121 gtccccagtg ctatctggga catattcctc cgcccagagc agggtcccgc ttccctaagg
    12181 ccctgctctg ggcttctggg tttgagtcct tggcaagccc aggagaggcg ctcaggcttc
    12241 cctgtccccc ttcctcgtcc accatctcat gcccctggct ctcctgcccc ttccctacag
    12301 gggttcctgg ctctgctcta agggcgaatt cgcggccgct aaattcaatt cgccctatag
    12361 tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
    12421 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
    12481 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctatacgtac ggcagtttaa
    12541 ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat
    12601 tattgacacg ccggggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga
    12661 taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat
    12721 gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg ctgatctcag
    12781 ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc
    12841 aggcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttcacgtaga aagccagtcc
    12901 gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga caagggaaaa
    12961 cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat agctagactg
    13021 ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct ctggtaaggt
    13081 tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct gatggcgcag
    13141 gggatcaagc tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
    13201 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
    13261 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
    13321 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
    13381 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
    13441 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
    13501 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
    13561 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
    13621 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
    13681 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
    13741 gacccatgca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
    13801 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
    13861 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
    13921 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
    13981 gaattattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
    14041 tttcacaccg catcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
    14101 tttttctaaa tacattcaaa tatgtatccg ctcatgagat tatcaaaaag gatcttcacc
    14161 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
    14221 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
    14281 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
    14341 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
    14401 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
    14461 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
    14521 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
    14581 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
    14641 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
    14701 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
    14761 agatgctttt ctgtgactgg tgaatactca accaagtcat tctgagaata gtgtatgcgg
    14821 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
    14881 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
    14941 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
    15001 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
    15061 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
    15121 atttatcagg gttattgtct catgaccaaa atcccttaac gtgagttttc gttccactga
    15181 gcgtcagacc ccg
//
