LOCUS       sequence-493902-       13542 bp    DNA     circular SYN 25-NOV-2025
DEFINITION  For microglia differentiation from iPSC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13542)
  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 13542)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 25, 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: CYBL-SPI1-CEBPB-IRF5
FEATURES             Location/Qualifiers
     source          1..13542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(5..805)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="GPRIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(116..135)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     726..745
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(800..853)
                     /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    877..902
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    903..936
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             join(1794..1862,1863..2654)
                     /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(2012..2031)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2825..3413
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3314..3333
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4439..4457
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4475..4493
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4511..4529
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4547..4565
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4583..4601
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4619..4637
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4655..4673
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4718..4742
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4825..4834
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5650..5703
                     /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             6767..6820
                     /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    8334..8455
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8371..8390)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8425..8444
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8504..8520)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8504..8520)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8517..8539)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8528..8544
                     /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(8552..8558,8559..8576,8577..8582))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8597..8618
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8735..8752)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     9044..9063
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    9062..9117
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     9163..9182
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(9273..10019)
                     /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(10051..10070)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          10078..11094
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(10132..10154)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(11095..11372)
                     /label=chicken beta-actin promoter
     enhancer        11374..11753
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    11759..11983
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     11972..11989
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    12005..12038
                     /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(12314..12333)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     12583..12601
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      12743..12752
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             12790..13058
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(13071..13282)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     polyA_signal    13318..13542
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 gtttttagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg
       61 cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat
      121 cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga
      181 tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg
      241 tcacgacgag atcctcgccg tcgggcatgc gcgccttgag cctggcgaac agttcggctg
      301 gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc
      361 gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat
      421 caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa
      481 ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg
      541 cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata
      601 gccgcgctgc ctcgtcctgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa
      661 gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct
      721 gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca
      781 atccatcttg ttcaatccta gggccgggat tctcctccac gtcaccgcat gttagaagac
      841 ttcctctgcc ctctccgctg ccagatctct cgaggccctg tgggaggaag agaagaggtc
      901 agataacttc gtataatgta tgctatacga agttatgcgg ccgctgggag aagggtaaga
      961 gagctcaatc ctcatcttac atagaaggaa gtcagtagat aatatcttac atttaaaagt
     1021 caagaaataa cagtctgacc atactatcta gaaatatgga agttaacagt aaggaagtta
     1081 aaacagttga aagtagtttc gtctagaaag aaagacttgg ggaagaacaa agaaggctaa
     1141 ttttcattat gagctttatt gtaacagaaa agagaccact aactcagcta ggttttccca
     1201 attcctcaac actttggggc acagtgttta gtcagagctg tgatcactct ctgagcacat
     1261 ggattgtatc ttaattttca tctcattgaa aatatgtacc atttgtccaa gtggaaactg
     1321 ggctgaagca tgactgtgac caccaccaaa gaaatggcta gactcgctaa ggtctgctgg
     1381 ggtgacccgt gattgtgttg tgcttgatcc tgttgagcat ggctctttta agtcaccata
     1441 cctgcccctg ccaatgcacc aaacaccaac ttaaagggac tttcagcttt tgtaagtcag
     1501 tctcagcctt acttctgaac catttctttg acttctttca tgactctgat cctgtcccag
     1561 ttttaagcct cccagccacc agtcatgctg ttttgctctg ctatttacat tcttctctac
     1621 ctactttgga cttgatggta tgcctactat attagttatt tattgctgtg taacaaacca
     1681 ctctgaaaca tagtggctta caaccacaac cattttattt tattatgaaa ctgtgggtca
     1741 gaatgtatat tggcccattg gcatgattct cctgctattg aaaaaggaag agtatgagta
     1801 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1861 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1921 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1981 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     2041 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2101 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2161 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     2221 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2281 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     2341 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     2401 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     2461 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     2521 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     2581 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     2641 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     2701 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2761 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2821 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2881 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2941 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3001 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3061 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3121 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3181 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3241 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3301 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3361 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3421 gcaacgcggc ctttttacgg ttcctggcga attcgccctt agagcagagc caggaacccc
     3481 tgtagggaag gggcaggaga gccaggggca tgagatggtg gacgaggaag ggggacaggg
     3541 aagcctgagc gcctctcctg ggcttgccaa ggactcaaac ccagaagccc agagcagggc
     3601 cttagggaag cgggacggct ctgttggaga gtccagtatt gaattagtga tagatacaca
     3661 gaaaaaaagc aaattgaata ggtgaatgga taaacaaact gtggaacatc cagacaatga
     3721 aatattcacc aataaaaaga aatgagctat caagccacat tggctatgcc cttcacattt
     3781 aaggaggaac cttaaatgca tattgccaaa ttttaaaagc tagtctgaaa atgctacaga
     3841 ctgtatgatt ccaattatat gacattttag aaaaggcaaa actatagaga tcagtggttg
     3901 ccagcggctc atggaggcag gatgaattgg tggaacacag ggatgctttc agggtaatga
     3961 aaatatcttg tgtgatcctg taataatgga tacgtggcat aatgcatttg ttaaaacctg
     4021 taaactagcc aggtatagtg gcttttttgc acctatagtc tcagctactt ggtaggctga
     4081 gacaggaaga tcacttgagc ccaggacttc aagaccagcc tgggtaatat agtaagaccc
     4141 catctctatt ttttaaagaa attaagaaag aaaagaaacc catcaactgt ataacacaaa
     4201 gaatcgtaat gtaaactaca gtcctgagta atgaaacatt tttaaagaaa taaataaatt
     4261 caaggcaatt attaacttca ggaaaaacag taaagtaaac ataagtatac aaatcaaaca
     4321 tggctcagtt gtgaaaatat ttacaaggtc actttaagta acattgaatg ttgatataat
     4381 cccaaaatat atttatgttg gaaggatgag gagatatctc caccccacag tggtttactc
     4441 cctatcagtg atagagaacg tatgaagagt ttactcccta tcagtgatag agaacgtatg
     4501 cagactttac tccctatcag tgatagagaa cgtataagga gtttactccc tatcagtgat
     4561 agagaacgta tgaccagttt actccctatc agtgatagag aacgtatcta cagtttactc
     4621 cctatcagtg atagagaacg tatatccagt ttactcccta tcagtgatag agaacgtata
     4681 agctttaggc gtgtacggtg ggcgcctata aaagcagagc tcgtttagtg aaccgtcaga
     4741 tcgcctggag caattccaca acacttttgt cttataccaa ctttccgtac cacttcctac
     4801 cctcgtaaac ttaaggttaa ttaagccacc atggaaggat tccccctggt gccccctccc
     4861 tccgaagatc tcgtgcccta cgacaccgat ctgtaccaga gacagacaca tgagtattac
     4921 ccctacctgt ccagcgacgg agagtcccac agcgaccact attgggattt tcacccccac
     4981 catgtgcact ccgagttcga gagctttgct gagaacaact tcaccgagct ccagtccgtg
     5041 cagcctcctc aactccagca gctctataga cacatggagc tggagcagat gcatgtgctg
     5101 gataccccca tggtgcctcc ccaccctagc ctgggacacc aagtctccta cctgcccagg
     5161 atgtgcctcc agtacccttc cctgtccccc gctcagccca gcagcgatga ggaagaaggc
     5221 gaaaggcagt cccctcctct ggaggtgtcc gacggagagg ctgatggcct cgagcctgga
     5281 cccggactgc tgcccggcga gaccggcagc aaaaagaaga ttaggctgta ccagttcctc
     5341 ctggatctgc tcagatccgg cgacatgaag gacagcattt ggtgggtcga caaggacaag
     5401 ggcacctttc agttctccag caagcacaaa gaggctctgg ctcacaggtg gggcatccag
     5461 aagggcaaca ggaagaagat gacctatcag aagatggcca gggccctcag aaactacggc
     5521 aagaccggag aggtgaaaaa agtcaagaag aagctcacat accaattctc cggcgaggtg
     5581 ctgggaagag gaggactggc tgagaggaga caccctcctc actccaggat gcatagcagg
     5641 ggcagcggcg agggaagggg ttctctgctc acctgtggag atgtcgagga gaaccccggc
     5701 cccatgcaga ggctggtggc ttgggaccct gcctgcctcc ctctccctcc tccccctcct
     5761 gctttcaaga gcatggaggt cgccaacttt tactacgagg ccgattgtct ggccgccgct
     5821 tatggcggaa aagctgcccc cgccgccccc cccgccgcca gacccggccc caggcccccc
     5881 gctggcgaac tgggaagcat cggcgaccac gagagggcca tcgactttag cccctacctg
     5941 gagcctctcg gcgcccccca agcccccgcc cctgccacag ctaccgacac attcgaggcc
     6001 gccccccctg cccccgctcc cgcccccgcc tccagcggcc agcaccatga ctttctgtcc
     6061 gacctgttct ccgatgacta cggcggcaaa aactgcaaga aacccgccga gtacggctat
     6121 gtgtccctgg gcaggctggg agccgctaaa ggagccctgc accctggctg ttttgcccct
     6181 ctgcaccctc ctcctccccc cccccccccc ccccgccgag ctcaaagctg agcctggctt
     6241 cgagcccgcc gattgcaaga gaaaagagga ggctggcgcc cctggaggag gcgccggcat
     6301 ggctgctgga ttcccctatg ctctgagggc ctacctgggc taccaagccg tccctagcgg
     6361 aagcagcggc tccctgtcca catcctcctc ctccagccct cctggaacac cttcccccgc
     6421 cgatgctaaa gcccccccca cagcctgtta cgccggcgcc gctcccgccc cttcccaagt
     6481 gaaaagcaag gccaagaaga ccgtggacaa gcactccgac gagtataaaa tcagaaggga
     6541 aagaaacaac atcgccgtca gaaagtccag ggataaggcc aagatgagga acctcgagac
     6601 acagcacaag gtgctcgagc tcaccgccga gaacgaaaga ctgcaaaaga aagtcgagca
     6661 gctctccagg gaactgtcca ccctcagaaa cctgttcaaa cagctgcccg agcccctcct
     6721 ggccagcagc ggccactgtg gaagcatgca ttccagaggt tctggcgaag gaaggggcag
     6781 cctgctgaca tgcggcgacg tggaagaaaa tcccggcccc atgaatcaga gcattcccgt
     6841 ggcccctacc ccccctagaa gagtcagact gaaaccttgg ctcgtcgccc aggtgaatag
     6901 ctgtcagtac cctggcctgc aatgggtgaa cggagagaag aagctctttt gcatcccctg
     6961 gagacacgct accaggcacg gaccttccca ggatggcgac aacaccattt tcaaggcctg
     7021 ggccaaggag accggaaaat acaccgaggg cgtcgacgaa gccgaccctg ccaagtggaa
     7081 agccaacctc agatgtgccc tcaacaagag cagggatttc aggctgattt acgacggccc
     7141 cagggacatg cctcctcaac cttacaaaat ctatgaggtg tgcagcaatg gccctgcccc
     7201 cacagacagc cagcctcccg aggactattc cttcggagcc ggagaagagg aagaagaaga
     7261 agaggagctc cagaggatgc tccccagcct gagcctcacc gaagacgtga agtggccccc
     7321 tacactccag cctcccacac tgaggccccc cacactccaa ccccctacac tgcagccccc
     7381 cgtggtgctg ggcccccccg cccccgaccc cagccctctg gctcctcctc ccggcaatcc
     7441 tgccggcttt agggaactgc tgagcgaagt cctggagccc ggccccctgc ctgctagcct
     7501 gccccctgct ggagagcagc tgctgcctga tctcctcatt agcccccaca tgctgcccct
     7561 caccgatctg gagatcaagt tccagtacag aggcaggccc cccagagccc tgaccatctc
     7621 caaccctcac ggctgtaggc tcttctactc ccagctggag gccacacaag agcaggtgga
     7681 actgttcggc cccatcagcc tggaacaggt gaggttcccc tcccccgagg atatcccctc
     7741 cgacaagcag aggttttaca ccaatcagct cctggatgtg ctggacagag gcctgatcct
     7801 gcagctgcag ggccaggatc tctacgctat caggctctgc cagtgcaagg tgttttggag
     7861 cggaccttgt gctagcgccc acgactcctg tcccaacccc attcaaagag aggtgaagac
     7921 caaactcttc agcctcgagc acttcctcaa tgagctcatc ctgttccaga aaggccaaac
     7981 caataccccc ccccctttcg agatcttctt ctgcttcgga gaggagtggc ctgacaggaa
     8041 gcccagagaa aaaaagctca ttacagtgca ggtcgtgccc gtcgctgcca gactcctcct
     8101 ggagatgttt agcggcgagc tcagctggag cgccgacagc atcaggctgc agatcagcaa
     8161 ccccgacctc aaagatagga tggtggagca gtttaaagag ctccaccaca tctggcaatc
     8221 ccagcagaga ctgcagcctg tcgctcaggc tccccccggc gccggcctgg gcgtgggcca
     8281 aggcccctgg cctatgcacc ctgctggaat gcagtgaatc gatcgcccac cccaacttgt
     8341 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     8401 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     8461 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     8521 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     8581 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     8641 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     8701 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     8761 tcgttcggct gcggcgagcg gtatcagctc actcaaagga cggccgcaca tgtaaggaaa
     8821 attttaggga tgttaaagaa aaaaataaca caaaacaaaa tataaaaaaa atctaacctc
     8881 aagtcaaggc ttttctatgg aataaggaat ggacagcagg gggctgtttc atatactgat
     8941 gacctcttta tagccaacct ttgttcatgg cagccagcat atgggcatat gttgccaaac
     9001 tctaaaccaa atactcattc tgatgtttta aatgatttgc cctcccatat gtccttccga
     9061 gtgagagaca caaaaaattc caacacacta ttgcaatgaa aataaatttc ctttattagc
     9121 cagaagtcag atgctcaagg ggcttcatga tgtccccata atttttggca gagggaaaaa
     9181 gatctcagtg gtatttgtga gccagggcat tggccacacc agtcaccacc ttctgatagg
     9241 cagcctgcac ctgaggagtg aattcacgcg ttttagcctg gcagcatatc gagatcgaag
     9301 tcatcgagag cgtcagcagg gagcatatcc agatcaaaat catccagggc atcggcgggc
     9361 agcatgtcca ggtcgaagtc gtccagggcg tcggtgggtc cgccgctctc gcacttcagc
     9421 tgcttctcga ggccgcagat gatcagttcc aggccgaaca ggaaggcggg ctcggcgccc
     9481 tgtctgtcga acagctcgat ggcctgcttc agcagggggg gcatgctgtc ggtggtgggt
     9541 gtctctcttt cctctttggc gacctggtgt tcctgttctt ccagcacgca gcccagggtg
     9601 aagtggccca cggcgctcag ggcgtacagg gcgttttcca ggctgaagcc ctgctggcac
     9661 aggaaggcca gctggttttc cagtgtctcg tactgcttct cggtgggtct ggtgcccagg
     9721 tgcactttgg cgccgtcccg gtggctcagc agggcgcatc tgtagctctt ggcgttgttc
     9781 cgcaggaagt cctgccagct ctcgccttcc agagggcagc tgtgggtgtg gtgccggtcc
     9841 agcatctcga tgggcagggc gtccagcagg gcccgcttgt tcttcacgtg ccagtacagg
     9901 gtgggctgtt ccacgcccag tttctgggcc agcttccggg tggtcaggcc ctcgatgccc
     9961 acgccgttca gcagttccag ggcgctgttg atcactttgc tcttgtccag ccggctcatg
    10021 tcgacattct ttgccaaaat gatgagacag cacaataacc agcacgttgc ccaggagctg
    10081 taggaaaaag aagaaggcat gaacatggtt agcagaggct ctagagccgc cggtcacacg
    10141 ccagaagccg aaccccgccc tgccccgtcc cccccgaagg cagccgtccc cccgcggaca
    10201 gccccgaggc tggagaggga gaaggggacg gcggcgcggc gacgcacgaa ggccctcccc
    10261 gcccatttcc ttcctgccgg cgccgcaccg cttcgccccg cgcccgctag agggggtgcg
    10321 gcggcgcctc ccagatttcg gctccgcaca gatttgggac aaaggaagtc cctgcgccct
    10381 ctcgcacgat taccataaaa ggcaatggct gcggctcgcc gcgcctcgac agccgccggc
    10441 gctccggggg ccgccgcgcc cctcccccga gccctccccg gcccgaggcg gccccgcccc
    10501 gcccggcacc cccacctgcc gccacccccc gcccggcacg gcgagccccg cgccacgccc
    10561 cgtacggagc cccgcacccg aagccgggcc gtgctcagca actcggggag gggggtgcag
    10621 ggggggttgc agcccgaccg acgcgcccac accccctgct caccccccca cgcacacacc
    10681 ccgcacgcag cctttgttcc cctcgcagcc ccccccgcac cgcggggcac cgcccccggc
    10741 cgcgctcccc tcgcgcacac tgcggagcgc acaaagcccc gcgccgcgcc cgcagcgctc
    10801 acagccgccg ggcagcgcgg agccgcacgc ggcgctcccc acgcacacac acacgcacgc
    10861 accccccgag ccgctccccc cgcacaaagg gccctcccgg agcccctcaa ggctttcacg
    10921 cagccacaga aaagaaacaa gccgtcatta aaccaagcgc taattacagc ccggaggaga
    10981 agggccgtcc cgcccgctca cctgtgggag taacgcggtc agtcagagcc ggggcgggcg
    11041 gcgcgaggcg gcggcggagc ggggcacggg gcgaaggcag cgcgcagcga ctcccgcccg
    11101 ccgcgcgctt cgctttttat agggccgccg ccgccgccgc ctcgccataa aaggaaactt
    11161 tcggagcgcg ccgctctgat tggctgccgc cgcacctctc cgcctcgccc cgccccgccc
    11221 ctcgccccgc cccgccccgc ctggcgcgcg cccccccccc ccccccgccc ccatcgctgc
    11281 acaaaataat taaaaaataa ataaatacaa aattgggggt ggggaggggg gggagatggg
    11341 gagagtgaag cagaacgtgg ggctcacctc gaccatggta atagcgatga ctaatacgta
    11401 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
    11461 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
    11521 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
    11581 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
    11641 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
    11701 aactaatgac cccgtaattg attactatta ataactagtc aataatcaat gtcgagaccc
    11761 atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc ccccttgctg
    11821 tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt
    11881 tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg
    11941 ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaggc tgatcagcgg
    12001 gtttataact tcgtataatg tatgctatac gaagttatgg taccttactc gtccatgccg
    12061 ccggtggagt ggcggccctc ggcgcgttcg tactgttcca cgatggtgta gtcctcgttg
    12121 tgggaggtga tgtccaactt gatgttgacg ttgtaggcgc cgggcagctg cacgggcttc
    12181 ttggccttgt aggtggtctt gacctcagcg tcgtagtggc cgccgtcctt cagcttcagc
    12241 ctctgcttga tctcgccctt cagggcgccg tcctcggggt acatccgctc ggaggaggcc
    12301 tcccagccca tggtcttctt ctgcattacg gggccgtcgg aggggaagtt ggtgccgcgc
    12361 agcttcacct tgtagatgaa ctcgccgtcc tgcagggagg agtcctgggt cacggtcacc
    12421 acgccgccgt cctcgaagtt catcacgcgc tcccacttga agccctcggg gaaggacagc
    12481 ttcaagtagt cggggatgtc ggcggggtgc ttcacgtagg ccttggagcc gtacatgaac
    12541 tgaggggaca ggatgtccca ggcgaagggc agggggccac ccttggtcac cttcagcttg
    12601 gcggtctggg tgccctcgta ggggcggccc tcgccctcgc cctcgatctc gaactcgtgg
    12661 ccgttcacgg agccctccat gtgcaccttg aagcgcatga actccttgat gatggccatg
    12721 ttatcctcct cgcccttgct caccatggtg gcgaccggta gcgctagcgt aggcgccggt
    12781 cacagcttgg atctgtaacg gcgcagaaca gaaaacgaaa caaagacgta gagttgagca
    12841 agcagggtca ggcaaagcgt ggagagccgg ctgagtctag gtaggctcca agggagcgcc
    12901 ggacaaaggc ccggtctcga cctgagcttt aaacttacct agacggcgga cgcagttcag
    12961 gaggcaccac aggcgggagg cggcagaacg cgactcaacc ggcgtggatg gcggcctcag
    13021 gtagggcggc gggcgcgtga aggagagatg cgagcccctc gaagcttcag ctgtgttctg
    13081 gcggcaaacc cgttgcgaaa aagaacgttc acggcgacta ctgcacttat atacggttct
    13141 cccccaccct cgggaaaaag gcggagccag tacacgacat cactttccca gtttaccccg
    13201 cgccaccttc tctaggcacc ggttcaattg ccgacccctc cccccaactt ctcggggact
    13261 gtgggcgatg tgcgctctgc ccactgacgg gcaccggagc gatcgcaaat catgcatcca
    13321 tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt
    13381 cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt
    13441 cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg
    13501 gggaggggca aacaacagat ggctggcaac tagaaggcac ag
//
