LOCUS       sequence-318410-       11546 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for genetic tracing of mesenchymal Glioblastoma
            cell-state..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11546)
  AUTHORS   Schmitt MJ, Company C, Dramaretska Y, Barozzi I, Gohrig A, Kertalli 
            S, Grossmann M, Naumann H, Sanchez-Bailon MP, Hulsman D, Glass R, 
            Squatrito M, Serresi M, Gargiulo G
  TITLE     Phenotypic Mapping of Pathologic Cross-Talk between Glioblastoma and
            Innate Immune Cells by Synthetic Genetic Tracing.
  JOURNAL   Cancer Discov. 2021 Mar;11(3):754-777. doi: 
            10.1158/2159-8290.CD-20-0219. Epub 2020 Dec 23.
  PUBMED    33361384
REFERENCE   2  (bases 1 to 11546)
  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: 
            "10.1158/2159-8290.CD-20-0219"; journalName: "Cancer Discov"; date: 
            "2021-03"; volume: "11"; issue: "3"; pages: "754-777"
COMMENT     Sequence Label: FH1-MGT#2-mCherry_PGK-H2B-CFP
FEATURES             Location/Qualifiers
     source          1..11546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1052..1070
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1088..1106
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1156..1863
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1298..1316)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1566..1585
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1993..2581
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2046..2066)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2464..2475)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2584..2600)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2585..2601)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        2725..3235
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3154..3172
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3247..3624
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      3637..3646
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3643..3645,3646..3648,3649..4362)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3688..3709)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3949..3968)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4296..4317
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4753..4933
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4959..4976)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4965..5189
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5235..5663
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5322..5341)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5532..5553
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5672..5692)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5677..6006
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5857..5992
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5919..5938
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6054..6101
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6120..6494
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    6624..6745
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6661..6680)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6715..6734
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6794..6810)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6794..6810)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6807..6829)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6818..6834
                     /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)."
     promoter        complement(join(6842..6848,6849..6866,6867..6872))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6887..6908
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7025..7042)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7196..7784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7276..7295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7955..8746,8747..8815))
                     /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     8578..8597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8816..8920)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8995..9014)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        9186..9565
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9567..9765
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9720..9740
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             9783..9963
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    10010..10135
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10628..10861
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11046..11090
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    11388..11505
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 taatgcatct agaacagcag ggccacctcc ttctctcccc cgcgggcatg ggcccccacc
       61 cccactgccg gcagagtgct gaggactcgt gcaccatgag aacttctgac catgagaact
      121 ttgacttccg gatttggggg atctgcccag gtgaacacaa tgcaaggggc tgcatgacct
      181 accaggacag aactttcccc aattacaggg tgactcacag ccgcattggt gactcacttc
      241 aatgtgtcat ttccggctgc tgtgtgtgag cagtggacac gtgagggggg ggtgggtgag
      301 agagacaggc cacattgtgc aacagatctc tagagctttt tcatcttgca aaactgaaac
      361 tgtataccca tggaacaaca gctccctgct cccctccccc tcagctcctg ggtagtgaca
      421 tttcttgatt ctcagtaaac tatcacaaga acaaaaaacc aaacaccgca tattctcact
      481 cataggtggg aattgaacaa tgagatcaca tggacacagg aaggggaata tcacactctg
      541 gggactgtgg tggggtgggg ggagtgggga gggatagcac tgggatgtcc caagagaagg
      601 ggaagagggg gaggtgttag agaacttgtg tgttcaaccg aaacatgatg aaaacaggga
      661 aagcccccaa gatacctgtc attcccgatg atgtcagatt cagcaaattc aatgataaca
      721 aaacattatg aaaaaattag taattaaaat aatacagcaa tgtgtatgaa caaaataatc
      781 aatgaaagtg aaacctaata gtaattccac aaacttatta caaagctatt aatttaaaga
      841 gtagtggcaa ttgaaaacca caaccaacac cagtgcttac agcagcaata cttttactca
      901 gacttcctgt ttctggaact tgccttcttt tttgctgtgt ttatacttcc cttgtctgtg
      961 gttagataag tataaagccc tagatctaag cttctctgtc ttcctccctc cctcccttcc
     1021 tcttactctc attcattgta cactagttta ctccctatca gtgatagaga acgtatatcc
     1081 agtttactcc ctatcagtga tagagaattc atgtatggca ggaagaagcg gagacagcga
     1141 cgaagaaagc ttggggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     1201 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     1261 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     1321 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     1381 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     1441 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     1501 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1561 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1621 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1681 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1741 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     1801 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctatacaag
     1861 taaaccggta tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     1921 tgctttaaaa aacctcccac acgtacaagt aaagcggccg cgactctaca attcgatatc
     1981 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2041 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2101 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     2161 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     2221 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     2281 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     2341 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     2401 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     2461 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     2521 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     2581 catcgatacc gtcgacctcg atcgaggacc tagaaaaaca tggagcaatc acaagtagca
     2641 atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg
     2701 gttttccagt cacacctcag gtacggggtt ggggttgcgc cttttccaag gcagccctgg
     2761 gtttgcgcag ggacgcggct gctctgggcg tggttccggg aaacgcagcg gcgccgaccc
     2821 tgggtctcgc acattcttca cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct
     2881 tgtgggcccc ccggcgacgc ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt
     2941 cgcggcgtgc cggacgtgac aaacggaagc cgcacgtctc actagtaccc tcgcagacgg
     3001 acagcgccag ggagcaatgg cagcgcgccg accgcgatgg gctgtggcca atagcggctg
     3061 ctcagcaggg cgcgccgaga gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg
     3121 ggcggtagtg tgggccctgt tcctgcccgc gcggtgttcc gcattctgca agcctccgga
     3181 gcgcacgtcg gcagtcggct ccctcgttga ccgaatcacc gacctctctc cccaggggga
     3241 tctaccatgc cagagccagc gaagtctgct cccgccccga aaaagggctc caagaaggcg
     3301 gtgactaagg cgcagaagaa aggcggcaag aagcgcaagc gcagccgcaa ggagagctat
     3361 tccatctatg tgtacaaggt tctgaagcag gtccaccctg acaccggcat ttcgtccaag
     3421 gccatgggca tcatgaactc gtttgtgaac gacattttcg agcgcatcgc aggtgaggct
     3481 tcccgcctgg cgcattacaa caagcgctcg accatcacct ccagggagat ccagacggcc
     3541 gtgcgcctgc tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg tactaaggcc
     3601 atcaccaagt acaccagcgc taaggatcca ccggtcgcca ccatggtgag caagggcgag
     3661 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     3721 aggttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     3781 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     3841 tggggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     3901 tccgccatgc ccgaaggcta cgtccaggag cgtaccatct tcttcaagga cgacggcaac
     3961 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     4021 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     4081 atcagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggcccacttc
     4141 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     4201 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     4261 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     4321 gccgccggga tcactctcgg catggacgag ctgtacaagt aggtaccttt aagaccaatg
     4381 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     4441 ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc
     4501 tacttccctg attggcagaa ctacacacca gggccaggga tcagatatcc actgaccttt
     4561 ggatggtgct acaagctagt accagttgag caagagaagg tagaagaagc caatgaagga
     4621 gagaacaccc gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa
     4681 gtattagagt ggaggtttga cagccgccta gcatttcatc acatggcccg agagctgcat
     4741 ccggactgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     4801 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     4861 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     4921 aaaatctcta gcagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     4981 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     5041 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     5101 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     5161 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     5221 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     5281 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     5341 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     5401 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     5461 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     5521 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     5581 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     5641 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     5701 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5761 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5821 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     5881 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     5941 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     6001 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca
     6061 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca
     6121 tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg
     6181 agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac ttcgccggtg
     6241 tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca
     6301 acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag tggtcggagg
     6361 tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc ggcgagcagc
     6421 cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg
     6481 aggagcagga ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt
     6541 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     6601 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     6661 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6721 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     6781 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6841 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6901 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6961 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     7021 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     7081 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     7141 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     7201 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     7261 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     7321 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     7381 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     7441 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7501 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7561 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7621 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7681 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     7741 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     7801 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7861 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     7921 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7981 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     8041 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     8101 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     8161 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     8221 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     8281 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     8341 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     8401 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     8461 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     8521 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     8581 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     8641 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     8701 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     8761 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     8821 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     8881 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     8941 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     9001 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     9061 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     9121 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     9181 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     9241 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     9301 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     9361 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     9421 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     9481 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     9541 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     9601 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     9661 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     9721 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagcgcgt tttgcctgta
     9781 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     9841 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     9901 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     9961 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg
    10021 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
    10081 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
    10141 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
    10201 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
    10261 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
    10321 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
    10381 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga
    10441 gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag
    10501 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
    10561 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
    10621 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
    10681 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
    10741 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
    10801 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
    10861 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
    10921 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
    10981 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
    11041 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
    11101 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
    11161 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
    11221 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa
    11281 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg
    11341 cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa
    11401 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
    11461 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
    11521 gacagcagag atccagtttg gttaat
//
