LOCUS       sequence-378197-       13278 bp    DNA     circular SYN 14-APR-2025
DEFINITION  For microglia differentiation from iPSC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13278)
  AUTHORS   Drager NM, Sattler SM, Huang CT, Teter OM, Leng K, Hashemi SH, Hong 
            J, Aviles G, Clelland CD, Zhan L, Udeochu JC, Kodama L, Singleton 
            AB, Nalls MA, Ichida J, Ward ME, Faghri F, Gan L, Kampmann M
  TITLE     A CRISPRi/a platform in human iPSC-derived microglia uncovers 
            regulators of disease states.
  JOURNAL   Nat Neurosci. 2022 Aug 11. pii: 10.1038/s41593-022-01131-4. doi: 
            10.1038/s41593-022-01131-4.
  PUBMED    35953545
REFERENCE   2  (bases 1 to 13278)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Neurosci. 2022 Aug 11. pii: 10.1038/s41593-022-01131-4. doi: 
            10.1038/s41593-022-01131-4."
COMMENT     Sequence Label: AAVS1-MAFB-IRF8-CEBPA
FEATURES             Location/Qualifiers
     source          1..13278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(54..122,123..914)
                     /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(272..291)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1085..1673
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1574..1593
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1721..2557
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     protein_bind    2590..2608
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2626..2644
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2662..2680
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2698..2716
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2734..2752
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2770..2788
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2806..2824
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2869..2893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2976..2985
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3372..3395
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             3465..3482
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3978..4031
                     /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             5337..5390
                     /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"
     polyA_signal    6493..6614
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6530..6549)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6584..6603
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6663..6679)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6663..6679)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6676..6698)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6687..6703
                     /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(6711..6717,6718..6735,6736..6741))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6756..6777
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6894..6911)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7203..7222
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    7221..7276
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     7322..7341
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(7432..8178)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(8210..8229)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          8237..9253
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(8291..8313)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(9254..9531)
                     /label=chicken beta-actin promoter
     enhancer        9533..9912
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    9918..10142
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     10131..10148
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    10164..10197
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(10473..10492)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     10742..10760
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      10902..10911
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             10949..11217
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(11230..11441)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     polyA_signal    11477..11701
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(11722..12321)
                     /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(11805..11825)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     12302..12321
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(12331..12384)
                     /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"
     misc_feature    12408..12433
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    12440..12473
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    12475..13278
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
ORIGIN
        1 aagggcgaat tcaaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
       61 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      121 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      181 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      241 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
      301 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
      361 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
      421 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
      481 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
      541 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
      601 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
      661 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
      721 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
      781 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
      841 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
      901 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
      961 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1021 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1081 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1141 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1201 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1261 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1321 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1381 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     1441 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1501 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1561 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1621 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     1681 gcaacgcggc ctttttacgg ttcctggcga attcgccctt agagcagagc caggaacccc
     1741 tgtagggaag gggcaggaga gccaggggca tgagatggtg gacgaggaag ggggacaggg
     1801 aagcctgagc gcctctcctg ggcttgccaa ggactcaaac ccagaagccc agagcagggc
     1861 cttagggaag cgggaccctg ctctgggcgg aggaatatgt cccagatagc actggggact
     1921 ctttaaggaa agaaggatgg agaaagagaa agggagtaga ggcggccacg acctggtgaa
     1981 cacctaggac gcaccattct cacaaaggga gttttccaca cggacacccc cctcctcacc
     2041 acagccctgc caggacgggg ctggctactg gccttatctc acaggtaaaa ctgacgcacg
     2101 gaggaacaat ataaattggg gactagaaag gtgaagagcc aaagttagaa ctcaggacca
     2161 acttattctg attttgtttt tccaaactgc ttctcctctt gggaagtgta aggaagctgc
     2221 agcaccagga tcagtgaaac gcaccagaca gccgcgtcag agcagctcag gttctgggag
     2281 agggtagcgc agggtggcca ctgagaaccg ggcaggtcac gcatcccccc cttccctccc
     2341 accccctgcc aagctctccc tcccaggatc ctctctggct ccatcgtaag caaaccttag
     2401 aggttctggc aaggagagag atggctccag gaaatggggg tgtgtcacca gataaggaat
     2461 ctgcctaaca ggaggtgggg gttagaccca atatcaggag actaggaagg aggaggccta
     2521 aggatggggc ttttctgtca ccaatcctgt ccctagtaaa gctgatatct ccaccccaca
     2581 gtggtttact ccctatcagt gatagagaac gtatgaagag tttactccct atcagtgata
     2641 gagaacgtat gcagacttta ctccctatca gtgatagaga acgtataagg agtttactcc
     2701 ctatcagtga tagagaacgt atgaccagtt tactccctat cagtgataga gaacgtatct
     2761 acagtttact ccctatcagt gatagagaac gtatatccag tttactccct atcagtgata
     2821 gagaacgtat aagctttagg cgtgtacggt gggcgcctat aaaagcagag ctcgtttagt
     2881 gaaccgtcag atcgcctgga gcaattccac aacacttttg tcttatacca actttccgta
     2941 ccacttccta ccctcgtaaa cttaaggtta attaagccac catggccgct gagctgagca
     3001 tgggccctga gctccctacc tcccctctcg ccatggagta cgtgaacgac ttcgacctgc
     3061 tgaagttcga cgtcaaaaag gaacccctgg gcagggccga aaggcctgga aggccttgta
     3121 caaggctgca acccgccgga tccgtgtcca gcacacccct ctccacacct tgttccagcg
     3181 tgccttccag ccccagcttt tcccctaccg agcagaagac acacctggag gacctgtact
     3241 ggatggccag caactaccag cagatgaacc ccgaagctct gaatctgaca cctgaagacg
     3301 ctgtggaggc cctcattggc tcccaccccg tgcctcagcc cctgcagtcc tttgacagct
     3361 tcagaggcgc ccatcaccac catcatcatc accatcccca ccctcaccat gcttaccctg
     3421 gcgccggagt ggcccatgac gaactcggac ctcatgccca tcctcaccac caccatcacc
     3481 accaggcttc ccctcctcct tccagcgctg cttcccccgc tcagcaactc cctaccagcc
     3541 accccggacc tggccctcat gccacagcta gcgccacagc tgctggcggc aatggatccg
     3601 tggaggacag attcagcgac gatcagctgg tgagcatgtc cgtgagggaa ctgaacaggc
     3661 acctgagggg ctttaccaaa gatgaggtga tcaggctgaa gcagaagaga aggaccctca
     3721 aaaacagggg ctatgcccag agctgcagat ataagagggt ccaacaaaaa caccatctgg
     3781 agaatgagaa gacccagctg attcagcagg tggagcaact caagcaggag gtgagcagac
     3841 tcgccagaga gagggatgcc tataaagtca aatgtgagaa actggccaac tccggcttca
     3901 gggaggccgg aagcaccagc gattccccca gctcccctga gttctttctc agcaggatgc
     3961 actccagggg atccggagag ggcaggggct ccctgctgac atgcggcgac gtggaagaga
     4021 accccggccc catgtgcgac aggaatggag gcagaaggct gaggcagtgg ctgatcgagc
     4081 agattgattc cagcatgtat cccggcctga tctgggagaa tgaggagaag agcatgttca
     4141 ggatcccctg gaagcacgct ggcaagcagg actataatca ggaagtggac gcctccatct
     4201 ttaaggcctg ggccgtcttc aagggcaagt ttaaggaagg agacaaggct gaacccgcca
     4261 catggaagac cagactgagg tgcgccctga acaaatcccc cgattttgaa gaggtgaccg
     4321 acaggtccca gctggacatt tccgagcctt acaaagtcta taggatcgtg cccgaggagg
     4381 aacaaaagtg taagctcgga gtcgctaccg ctggatgcgt gaacgaggtc acagagatgg
     4441 agtgcggcag gagcgagatc gacgaactca tcaaggaacc tagcgtggat gactacatgg
     4501 gcatgatcaa aaggagccct agcccccctg aggcctgcag aagccagctc ctgcctgatt
     4561 ggtgggccca gcaacccagc acaggcgtcc ctctcgtgac cggctacacc acctatgatg
     4621 cccaccattc cgccttcagc cagatggtga tctccttcta ctacggcgga aagctcgtgg
     4681 gacaggccac caccacctgt cccgaaggct gtaggctcag cctgtcccag cctggcctgc
     4741 ccggaaccaa actgtacgga cctgagggac tggagctcgt gagatttccc cctgccgatg
     4801 ccatccccag cgagagacag aggcaggtga ccagaaagct gttcggccac ctcgaaaggg
     4861 gcgtcctgct ccactcctcc agacagggcg tcttcgtgaa gaggctctgc cagggaagag
     4921 tgttttgcag cggaaacgct gtggtgtgta aaggcagacc caacaagctc gagagggacg
     4981 aggtggtgca ggtgttcgac accagccagt tctttaggga gctccaacag ttctacaaca
     5041 gccagggcag actccctgat ggcagggtcg tcctgtgctt cggagaggag tttcccgata
     5101 tggctcctct cagatccaag ctgatcctgg tccagattga gcagctgtac gtgagacagc
     5161 tggccgagga agctggcaag agctgtggag ccggcagcgt gatgcaggct cccgaggaac
     5221 ctcctcctga ccaagtgttc aggatgttcc ccgatatttg tgcttcccac caaaggtcct
     5281 ttttcaggga gaaccagcag atcaccgtgg tcaacatgca cagcagggga tccggcgagg
     5341 gaaggggaag cctgctgaca tgtggcgatg tggaggagaa ccctggccct atggagagcg
     5401 ctgatttcta cgaagccgaa cccagacctc ccatgtccag ccacctccag agccctcccc
     5461 atgctcccag ctccgctgcc tttggatttc ctagaggcgc tggacctgcc cagcctcctg
     5521 cccctcccgc cgctcccgag cccctgggcg gcatttgcga acacgaaacc tccatcgaca
     5581 tctccgccta catcgacccc gccgctttca acgacgagtt tctggccgac ctgttccaac
     5641 acagcaggca gcaggaaaag gccaaggccg ccgtcggacc cacaggcggc ggcggcggag
     5701 gagacttcga ctatcctggc gctcccgccg gccctggcgg cgctgtgatg cctggaggag
     5761 ctcacggacc tcctcctggc tatggctgcg ccgctgccgg atatctggac ggcagactgg
     5821 aacctctcta tgagagggtc ggagctcccg ccctcagacc tctggtgatc aaacaggagc
     5881 ctagggagga ggacgaggcc aaacagctcg ccctggccgg cctgttccct tatcagcccc
     5941 ctcctcctcc ccctcctagc caccctcacc cccatcctcc tcccgctcat ctcgctgccc
     6001 ctcacctgca attccaaatc gctcattgcg gccagaccac catgcacctg caacccggcc
     6061 accccacccc tccccctaca cctgtccctt cccctcaccc cgctcctgct ctgggagccg
     6121 ccggactgcc tggccctgga tccgctctga aaggactggg agccgctcac cctgacctca
     6181 gagctagcgg cggaagcgga gctggcaagg ctaagaagag cgtcgataag aacagcaacg
     6241 aatacagggt gaggagagag agaaataaca tcgccgtcag gaagtccaga gacaaggcta
     6301 agcagagaaa tgtggagacc cagcagaaag tcctggaact cacatccgac aacgacaggc
     6361 tcagaaagag ggtcgagcaa ctctccaggg agctggacac cctgagagga attttcaggc
     6421 agctgcccga aagctccctc gtcaaggcca tgggcaattg tgcctgagcg gccgcaatcg
     6481 atcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6541 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6601 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     6661 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6721 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6781 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6841 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6901 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggac
     6961 ggccgcacat gtaaggaaaa ttttagggat gttaaagaaa aaaataacac aaaacaaaat
     7021 ataaaaaaaa tctaacctca agtcaaggct tttctatgga ataaggaatg gacagcaggg
     7081 ggctgtttca tatactgatg acctctttat agccaacctt tgttcatggc agccagcata
     7141 tgggcatatg ttgccaaact ctaaaccaaa tactcattct gatgttttaa atgatttgcc
     7201 ctcccatatg tccttccgag tgagagacac aaaaaattcc aacacactat tgcaatgaaa
     7261 ataaatttcc tttattagcc agaagtcaga tgctcaaggg gcttcatgat gtccccataa
     7321 tttttggcag agggaaaaag atctcagtgg tatttgtgag ccagggcatt ggccacacca
     7381 gccaccacct tctgataggc agcctgcacc tgaggagtga attcacgcgt tttagcctgg
     7441 cagcatatcg agatcgaagt catcgagagc gtcagcaggg agcatatcca gatcaaaatc
     7501 atccagggca tcggcgggca gcatgtccag gtcgaagtcg tccagggcgt cggtgggtcc
     7561 gccgctctcg cacttcagct gcttctcgag gccgcagatg atcagttcca ggccgaacag
     7621 gaaggcgggc tcggcgccct gtctgtcgaa cagctcgatg gcctgcttca gcaggggggg
     7681 catgctgtcg gtggtgggtg tctctctttc ctctttggcg acctggtgtt cctgttcttc
     7741 cagcacgcag cccagggtga agtggcccac ggcgctcagg gcgtacaggg cgttttccag
     7801 gctgaagccc tgctggcaca ggaaggccag ctggttttcc agtgtctcgt actgcttctc
     7861 ggtgggtctg gtgcccaggt gcactttggc gccgtcccgg tggctcagca gggcgcatct
     7921 gtagctcttg gcgttgttcc gcaggaagtc ctgccagctc tcgccttcca gagggcagct
     7981 gtgggtgtgg tgccggtcca gcatctcgat gggcagggcg tccagcaggg cccgcttgtt
     8041 cttcacgtgc cagtacaggg tgggctgttc cacgcccagt ttctgggcca gcttccgggt
     8101 ggtcaggccc tcgatgccca cgccgttcag cagttccagg gcgctgttga tcactttgct
     8161 cttgtccagc cggctcatgt cgacattctt tgccaaaatg atgagacagc acaataacca
     8221 gcacgttgcc caggagctgt aggaaaaaga agaaggcatg aacatggtta gcagaggctc
     8281 tagagccgcc ggtcacacgc cagaagccga accccgccct gccccgtccc ccccgaaggc
     8341 agccgtcccc ccgcggacag ccccgaggct ggagagggag aaggggacgg cggcgcggcg
     8401 acgcacgaag gccctccccg cccatttcct tcctgccggc gccgcaccgc ttcgccccgc
     8461 gcccgctaga gggggtgcgg cggcgcctcc cagatttcgg ctccgcacag atttgggaca
     8521 aaggaagtcc ctgcgccctc tcgcacgatt accataaaag gcaatggctg cggctcgccg
     8581 cgcctcgaca gccgccggcg ctccgggggc cgccgcgccc ctcccccgag cccyccccgg
     8641 cccgaggcgg ccccgccccg cccggcaccc ccacctgccg ccaccccccg cccggcacgg
     8701 cgagccccgc gccacgcccc gtacggagcc ccgcacccga agccgggccg tgctcagcaa
     8761 ctcggggagg ggggtgcagg gggggttgca gcccgaccga cgcgcccaca ccccctgctc
     8821 acccccccac gcacacaccc cgcacgcagc ctttgttccc ctcgcagccc cccccgcacc
     8881 gcggggcacc gcccccggcc gcgctcccct cgcgcacact gcggagcgca caaagccccg
     8941 cgccgcgccc gcagcgctca cagccgccgg gcagcgcgga gccgcacgcg gcgctcccca
     9001 cgcacacaca cacgcacgca ccccccgagc cgctcccccc gcacaaaggg ccctcccgga
     9061 gcccctcaag gctttcacgc agccacagaa aagaaacaag ccgtcattaa accaagcgct
     9121 aattacagcc cggaggagaa gggccgtccc gcccgctcac ctgtgggagt aacgcggtca
     9181 gtcagagccg gggcgggcgg cgcgaggcgg cggcggagcg gggcacgggg cgaaggcagc
     9241 gcgcagcgac tcccgcccgc cgcgcgcttc gctttttata gggccgccgc cgccgccgcc
     9301 tcgccataaa aggaaacttt cggagcgcgc cgctctgatt ggctgccgcc gcacctctcc
     9361 gcctcscccc sccccscccc ycsccccscc ccsccccscc yggcgcgcgc cccccccccc
     9421 cccccgcccc catcgctgca caaaataatt aaaaaataaa taaatacaaa attgggggtg
     9481 gggagggggg ggagatgggg agagtgaagc agaacgtggg gctcacctcg accatggtaa
     9541 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact
     9601 gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca
     9661 tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac
     9721 gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg
     9781 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga
     9841 actccatata tgggctatga actaatgacc ccgtaattga ttactattaa taactagtca
     9901 ataatcaatg tcgagaccca tagagcccac cgcatcccca gcatgcctgc tattgtcttc
     9961 ccaatcctcc cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact
    10021 cagacaatgc gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc
    10081 cagggtcaag gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac
    10141 agtcgaggct gatcagcggg tttataactt cgtataatgt atgctatacg aagttatggt
    10201 accttactcg tccatgccgc cggtggagtg gcggccctcg gcgcgttcgt actgttccac
    10261 gatggtgtag tcctcgttgt gggaggtgat gtccaacttg atgttgacgt tgtaggcgcc
    10321 gggcagctgc acgggcttct tggccttgta ggtggtcttg acctcagcgt cgtagtggcc
    10381 gccgtccttc agcttcagcc tctgcttgat ctcgcccttc agggcgccgt cctcggggta
    10441 catccgctcg gaggaggcct cccagcccat ggtcttcttc tgcattacgg ggccgtcgga
    10501 ggggaagttg gtgccgcgca gcttcacctt gtagatgaac tcgccgtcct gcagggagga
    10561 gtcctgggtc acggtcacca cgccgccgtc ctcgaagttc atcacgcgct cccacttgaa
    10621 gccctcgggg aaggacagct tcaagtagtc ggggatgtcg gcggggtgct tcacgtaggc
    10681 cttggagccg tacatgaact gaggggacag gatgtcccag gcgaagggca gggggccacc
    10741 cttggtcacc ttcagcttgg cggtctgggt gccctcgtag gggcggccct cgccctcgcc
    10801 ctcgatctcg aactcgtggc cgttcacgga gccctccatg tgcaccttga agcgcatgaa
    10861 ctccttgatg atggccatgt tatcctcctc gcccttgctc accatggtgg cgaccggtag
    10921 cgctagcgta ggcgccggtc acagcttgga tctgtaacgg cgcagaacag aaaacgaaac
    10981 aaagacgtag agttgagcaa gcagggtcag gcaaagcgtg gagagccggc tgagtctagg
    11041 taggctccaa gggagcgccg gacaaaggcc cggtctcgac ctgagcttta aacttaccta
    11101 gacggcggac gcagttcagg aggcaccaca ggcgggaggc ggcagaacgc gactcaaccg
    11161 gcgtggatgg cggcctcagg tagggcggcg ggcgcgtgaa ggagagatgc gagcccctcg
    11221 aagcttcagc tgtgttctgg cggcaaaccc gttgcgaaaa agaacgttca cggcgactac
    11281 tgcacttata tacggttctc ccccaccctc gggaaaaagg cggagccagt acacgacatc
    11341 actttcccag tttaccccgc gccaccttct ctaggcaccg gttcaattgc cgacccctcc
    11401 ccccaacttc tcggggactg tgggcgatgt gcgctctgcc cactgacggg caccggagcg
    11461 atcgcagatc atgcatccat agagcccacc gcatccccag catgcctgct attgtcttcc
    11521 caatcctccc ccttgctgtc ctgccccacc ccacccccca gaatagaatg acacctactc
    11581 agacaatgcg atgcaatttc ctcattttat taggaaagga cagtgggagt ggcaccttcc
    11641 agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact agaaggcaca
    11701 ggtttaaacg ggccctctag atcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg
    11761 tccttcgggc acctcgacgt cggcggtgac ggtgaagccg agccgctcgt agaaggggag
    11821 gttgcggggc gcggaggtct ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac
    11881 tccggggagc acgacggcgc tgcccagacc cttgccctgg tggtcgggcg agacgccgac
    11941 ggtggccagg aaccacgcgg gctccttggg ccggtgcggc gccaggaggc cttccatctg
    12001 ttgctgcgcg gccagccggg aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa
    12061 caccgccccc gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc
    12121 cgcgacccac accttgccga tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc
    12181 ggtgacccgc tcgatgtggc ggtccggatc gacggtgtgg cgcgtggcgg ggtagtcggc
    12241 gaacgcggcg gcgagggtgc gtacggccct ggggacgtcg tcgcgggtgg cgaggcgcac
    12301 cgtgggcttg tactcggtca tctcgagcct agggccggga ttctcctcca cgtcaccgca
    12361 tgttagaaga cttcctctgc cctctccgct gccagatctc tcgaggccct gtgggaggaa
    12421 gagaagaggt cagaagctta taacttcgta taatgtatgc tatacgaagt tattgcccca
    12481 ctgtggggtg gaggggacag ataaaagtac ccagaaccag agccacatta accggccctg
    12541 ggaatataag gtggtcccag ctcggggaca caggatccct ggaggcagca aacatgctgt
    12601 cctgaagtgg acataggggc ccgggttgga ggaagaagac tagctgagct ctcggacccc
    12661 tggaagatgc catgacaggg ggctggaaga gctagcacag actagagagg taaggggggt
    12721 aggggagctg cccaaatgaa aggagtgaga ggtgacccga atccacagga gaacggggtg
    12781 tccaggcaaa gaaagcaaga ggatggagag gtggctaaag ccagggagac ggggtacttt
    12841 ggggttgtcc agaaaaacgg tgatgatgca ggcctacaag aaggggaggc gggacgcaag
    12901 ggagacatcc gtcggagaag gccatcctaa gaaacgagag atggcacagg ccccagaagg
    12961 agaaggaaaa gggaacccag cgagtgaaga cggcatgggg ttgggtgagg gaggagagat
    13021 gcccggagag gacccagaca cggggaggat ccgctcagag gacatcacgt ggtgcagcgc
    13081 cgagaaggaa gtgctccgga aagagcatcc ttgggcagca acacagcaga gagcaagggg
    13141 aagagggagt ggaggaagac ggaacctgaa ggaggcggca gggaaggatc tgggccagcc
    13201 gtagaggtga cccaggccac aagctgcaga cagaaagcgg cacaggccca ggggagagaa
    13261 tgctggtcag agaaagca
//
