LOCUS       sequence-486659-       13278 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 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 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: AAVS1-MAFB-IRF8-CEBPA
FEATURES             Location/Qualifiers
     source          1..13278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    637..1473
                     /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    1506..1524
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1542..1560
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1578..1596
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1614..1632
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1650..1668
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1686..1704
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1722..1740
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1785..1809
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1892..1901
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2288..2311
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             2381..2398
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2894..2947
                     /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             4253..4306
                     /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    5409..5530
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5446..5465)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5500..5519
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5579..5595)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5579..5595)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5592..5614)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5603..5619
                     /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(5627..5633,5634..5651,5652..5657))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5672..5693
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5810..5827)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6119..6138
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    6137..6192
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     6238..6257
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(6348..7094)
                     /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(7126..7145)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          7153..8169
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(7207..7229)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(8170..8447)
                     /label=chicken beta-actin promoter
     enhancer        8449..8828
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    8834..9058
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     9047..9064
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    9080..9113
                     /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(9389..9408)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     9658..9676
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      9818..9827
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             9865..10133
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(10146..10357)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     polyA_signal    10393..10617
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(10638..11237)
                     /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(10721..10741)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     11218..11237
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(11247..11300)
                     /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    11324..11349
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    11356..11389
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    11391..12194
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     CDS             join(12248..12316,12317..13108)
                     /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(12466..12485)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggcgaattc gcccttagag cagagccagg aacccctgta
      661 gggaaggggc aggagagcca ggggcatgag atggtggacg aggaaggggg acagggaagc
      721 ctgagcgcct ctcctgggct tgccaaggac tcaaacccag aagcccagag cagggcctta
      781 gggaagcggg accctgctct gggcggagga atatgtccca gatagcactg gggactcttt
      841 aaggaaagaa ggatggagaa agagaaaggg agtagaggcg gccacgacct ggtgaacacc
      901 taggacgcac cattctcaca aagggagttt tccacacgga cacccccctc ctcaccacag
      961 ccctgccagg acggggctgg ctactggcct tatctcacag gtaaaactga cgcacggagg
     1021 aacaatataa attggggact agaaaggtga agagccaaag ttagaactca ggaccaactt
     1081 attctgattt tgtttttcca aactgcttct cctcttggga agtgtaagga agctgcagca
     1141 ccaggatcag tgaaacgcac cagacagccg cgtcagagca gctcaggttc tgggagaggg
     1201 tagcgcaggg tggccactga gaaccgggca ggtcacgcat cccccccttc cctcccaccc
     1261 cctgccaagc tctccctccc aggatcctct ctggctccat cgtaagcaaa ccttagaggt
     1321 tctggcaagg agagagatgg ctccaggaaa tgggggtgtg tcaccagata aggaatctgc
     1381 ctaacaggag gtgggggtta gacccaatat caggagacta ggaaggagga ggcctaagga
     1441 tggggctttt ctgtcaccaa tcctgtccct agtaaagctg atatctccac cccacagtgg
     1501 tttactccct atcagtgata gagaacgtat gaagagttta ctccctatca gtgatagaga
     1561 acgtatgcag actttactcc ctatcagtga tagagaacgt ataaggagtt tactccctat
     1621 cagtgataga gaacgtatga ccagtttact ccctatcagt gatagagaac gtatctacag
     1681 tttactccct atcagtgata gagaacgtat atccagttta ctccctatca gtgatagaga
     1741 acgtataagc tttaggcgtg tacggtgggc gcctataaaa gcagagctcg tttagtgaac
     1801 cgtcagatcg cctggagcaa ttccacaaca cttttgtctt ataccaactt tccgtaccac
     1861 ttcctaccct cgtaaactta aggttaatta agccaccatg gccgctgagc tgagcatggg
     1921 ccctgagctc cctacctccc ctctcgccat ggagtacgtg aacgacttcg acctgctgaa
     1981 gttcgacgtc aaaaaggaac ccctgggcag ggccgaaagg cctggaaggc cttgtacaag
     2041 gctgcaaccc gccggatccg tgtccagcac acccctctcc acaccttgtt ccagcgtgcc
     2101 ttccagcccc agcttttccc ctaccgagca gaagacacac ctggaggacc tgtactggat
     2161 ggccagcaac taccagcaga tgaaccccga agctctgaat ctgacacctg aagacgctgt
     2221 ggaggccctc attggctccc accccgtgcc tcagcccctg cagtcctttg acagcttcag
     2281 aggcgcccat caccaccatc atcatcacca tccccaccct caccatgctt accctggcgc
     2341 cggagtggcc catgacgaac tcggacctca tgcccatcct caccaccacc atcaccacca
     2401 ggcttcccct cctccttcca gcgctgcttc ccccgctcag caactcccta ccagccaccc
     2461 cggacctggc cctcatgcca cagctagcgc cacagctgct ggcggcaatg gatccgtgga
     2521 ggacagattc agcgacgatc agctggtgag catgtccgtg agggaactga acaggcacct
     2581 gaggggcttt accaaagatg aggtgatcag gctgaagcag aagagaagga ccctcaaaaa
     2641 caggggctat gcccagagct gcagatataa gagggtccaa caaaaacacc atctggagaa
     2701 tgagaagacc cagctgattc agcaggtgga gcaactcaag caggaggtga gcagactcgc
     2761 cagagagagg gatgcctata aagtcaaatg tgagaaactg gccaactccg gcttcaggga
     2821 ggccggaagc accagcgatt cccccagctc ccctgagttc tttctcagca ggatgcactc
     2881 caggggatcc ggagagggca ggggctccct gctgacatgc ggcgacgtgg aagagaaccc
     2941 cggccccatg tgcgacagga atggaggcag aaggctgagg cagtggctga tcgagcagat
     3001 tgattccagc atgtatcccg gcctgatctg ggagaatgag gagaagagca tgttcaggat
     3061 cccctggaag cacgctggca agcaggacta taatcaggaa gtggacgcct ccatctttaa
     3121 ggcctgggcc gtcttcaagg gcaagtttaa ggaaggagac aaggctgaac ccgccacatg
     3181 gaagaccaga ctgaggtgcg ccctgaacaa atcccccgat tttgaagagg tgaccgacag
     3241 gtcccagctg gacatttccg agccttacaa agtctatagg atcgtgcccg aggaggaaca
     3301 aaagtgtaag ctcggagtcg ctaccgctgg atgcgtgaac gaggtcacag agatggagtg
     3361 cggcaggagc gagatcgacg aactcatcaa ggaacctagc gtggatgact acatgggcat
     3421 gatcaaaagg agccctagcc cccctgaggc ctgcagaagc cagctcctgc ctgattggtg
     3481 ggcccagcaa cccagcacag gcgtccctct cgtgaccggc tacaccacct atgatgccca
     3541 ccattccgcc ttcagccaga tggtgatctc cttctactac ggcggaaagc tcgtgggaca
     3601 ggccaccacc acctgtcccg aaggctgtag gctcagcctg tcccagcctg gcctgcccgg
     3661 aaccaaactg tacggacctg agggactgga gctcgtgaga tttccccctg ccgatgccat
     3721 ccccagcgag agacagaggc aggtgaccag aaagctgttc ggccacctcg aaaggggcgt
     3781 cctgctccac tcctccagac agggcgtctt cgtgaagagg ctctgccagg gaagagtgtt
     3841 ttgcagcgga aacgctgtgg tgtgtaaagg cagacccaac aagctcgaga gggacgaggt
     3901 ggtgcaggtg ttcgacacca gccagttctt tagggagctc caacagttct acaacagcca
     3961 gggcagactc cctgatggca gggtcgtcct gtgcttcgga gaggagtttc ccgatatggc
     4021 tcctctcaga tccaagctga tcctggtcca gattgagcag ctgtacgtga gacagctggc
     4081 cgaggaagct ggcaagagct gtggagccgg cagcgtgatg caggctcccg aggaacctcc
     4141 tcctgaccaa gtgttcagga tgttccccga tatttgtgct tcccaccaaa ggtccttttt
     4201 cagggagaac cagcagatca ccgtggtcaa catgcacagc aggggatccg gcgagggaag
     4261 gggaagcctg ctgacatgtg gcgatgtgga ggagaaccct ggccctatgg agagcgctga
     4321 tttctacgaa gccgaaccca gacctcccat gtccagccac ctccagagcc ctccccatgc
     4381 tcccagctcc gctgcctttg gatttcctag aggcgctgga cctgcccagc ctcctgcccc
     4441 tcccgccgct cccgagcccc tgggcggcat ttgcgaacac gaaacctcca tcgacatctc
     4501 cgcctacatc gaccccgccg ctttcaacga cgagtttctg gccgacctgt tccaacacag
     4561 caggcagcag gaaaaggcca aggccgccgt cggacccaca ggcggcggcg gcggaggaga
     4621 cttcgactat cctggcgctc ccgccggccc tggcggcgct gtgatgcctg gaggagctca
     4681 cggacctcct cctggctatg gctgcgccgc tgccggatat ctggacggca gactggaacc
     4741 tctctatgag agggtcggag ctcccgccct cagacctctg gtgatcaaac aggagcctag
     4801 ggaggaggac gaggccaaac agctcgccct ggccggcctg ttcccttatc agccccctcc
     4861 tcctccccct cctagccacc ctcaccccca tcctcctccc gctcatctcg ctgcccctca
     4921 cctgcaattc caaatcgctc attgcggcca gaccaccatg cacctgcaac ccggccaccc
     4981 cacccctccc cctacacctg tcccttcccc tcaccccgct cctgctctgg gagccgccgg
     5041 actgcctggc cctggatccg ctctgaaagg actgggagcc gctcaccctg acctcagagc
     5101 tagcggcgga agcggagctg gcaaggctaa gaagagcgtc gataagaaca gcaacgaata
     5161 cagggtgagg agagagagaa ataacatcgc cgtcaggaag tccagagaca aggctaagca
     5221 gagaaatgtg gagacccagc agaaagtcct ggaactcaca tccgacaacg acaggctcag
     5281 aaagagggtc gagcaactct ccagggagct ggacaccctg agaggaattt tcaggcagct
     5341 gcccgaaagc tccctcgtca aggccatggg caattgtgcc tgagcggccg caatcgatcg
     5401 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5461 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5521 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5581 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5641 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5701 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5761 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5821 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggacggcc
     5881 gcacatgtaa ggaaaatttt agggatgtta aagaaaaaaa taacacaaaa caaaatataa
     5941 aaaaaatcta acctcaagtc aaggcttttc tatggaataa ggaatggaca gcagggggct
     6001 gtttcatata ctgatgacct ctttatagcc aacctttgtt catggcagcc agcatatggg
     6061 catatgttgc caaactctaa accaaatact cattctgatg ttttaaatga tttgccctcc
     6121 catatgtcct tccgagtgag agacacaaaa aattccaaca cactattgca atgaaaataa
     6181 atttccttta ttagccagaa gtcagatgct caaggggctt catgatgtcc ccataatttt
     6241 tggcagaggg aaaaagatct cagtggtatt tgtgagccag ggcattggcc acaccagcca
     6301 ccaccttctg ataggcagcc tgcacctgag gagtgaattc acgcgtttta gcctggcagc
     6361 atatcgagat cgaagtcatc gagagcgtca gcagggagca tatccagatc aaaatcatcc
     6421 agggcatcgg cgggcagcat gtccaggtcg aagtcgtcca gggcgtcggt gggtccgccg
     6481 ctctcgcact tcagctgctt ctcgaggccg cagatgatca gttccaggcc gaacaggaag
     6541 gcgggctcgg cgccctgtct gtcgaacagc tcgatggcct gcttcagcag ggggggcatg
     6601 ctgtcggtgg tgggtgtctc tctttcctct ttggcgacct ggtgttcctg ttcttccagc
     6661 acgcagccca gggtgaagtg gcccacggcg ctcagggcgt acagggcgtt ttccaggctg
     6721 aagccctgct ggcacaggaa ggccagctgg ttttccagtg tctcgtactg cttctcggtg
     6781 ggtctggtgc ccaggtgcac tttggcgccg tcccggtggc tcagcagggc gcatctgtag
     6841 ctcttggcgt tgttccgcag gaagtcctgc cagctctcgc cttccagagg gcagctgtgg
     6901 gtgtggtgcc ggtccagcat ctcgatgggc agggcgtcca gcagggcccg cttgttcttc
     6961 acgtgccagt acagggtggg ctgttccacg cccagtttct gggccagctt ccgggtggtc
     7021 aggccctcga tgcccacgcc gttcagcagt tccagggcgc tgttgatcac tttgctcttg
     7081 tccagccggc tcatgtcgac attctttgcc aaaatgatga gacagcacaa taaccagcac
     7141 gttgcccagg agctgtagga aaaagaagaa ggcatgaaca tggttagcag aggctctaga
     7201 gccgccggtc acacgccaga agccgaaccc cgccctgccc cgtccccccc gaaggcagcc
     7261 gtccccccgc ggacagcccc gaggctggag agggagaagg ggacggcggc gcggcgacgc
     7321 acgaaggccc tccccgccca tttccttcct gccggcgccg caccgcttcg ccccgcgccc
     7381 gctagagggg gtgcggcggc gcctcccaga tttcggctcc gcacagattt gggacaaagg
     7441 aagtccctgc gccctctcgc acgattacca taaaaggcaa tggctgcggc tcgccgcgcc
     7501 tcgacagccg ccggcgctcc gggggccgcc gcgcccctcc cccgagccct ccccggcccg
     7561 aggcggcccc gccccgcccg gcacccccac ctgccgccac cccccgcccg gcacggcgag
     7621 ccccgcgcca cgccccgtac ggagccccgc acccgaagcc gggccgtgct cagcaactcg
     7681 gggagggggg tgcagggggg gttgcagccc gaccgacgcg cccacacccc ctgctcaccc
     7741 ccccacgcac acaccccgca cgcagccttt gttcccctcg cagccccccc cgcaccgcgg
     7801 ggcaccgccc ccggccgcgc tcccctcgcg cacactgcgg agcgcacaaa gccccgcgcc
     7861 gcgcccgcag cgctcacagc cgccgggcag cgcggagccg cacgcggcgc tccccacgca
     7921 cacacacacg cacgcacccc ccgagccgct ccccccgcac aaagggccct cccggagccc
     7981 ctcaaggctt tcacgcagcc acagaaaaga aacaagccgt cattaaacca agcgctaatt
     8041 acagcccgga ggagaagggc cgtcccgccc gctcacctgt gggagtaacg cggtcagtca
     8101 gagccggggc gggcggcgcg aggcggcggc ggagcggggc acggggcgaa ggcagcgcgc
     8161 agcgactccc gcccgccgcg cgcttcgctt tttatagggc cgccgccgcc gccgcctcgc
     8221 cataaaagga aactttcgga gcgcgccgct ctgattggct gccgccgcac ctctccgcct
     8281 cgccccgccc cgcccctcgc cccgccccgc cccgcctggc gcgcgccccc cccccccccc
     8341 cgcccccatc gctgcacaaa ataattaaaa aataaataaa tacaaaattg ggggtgggga
     8401 ggggggggag atggggagag tgaagcagaa cgtggggctc acctcgacca tggtaatagc
     8461 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     8521 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     8581 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     8641 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg
     8701 ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc
     8761 catatatggg ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa
     8821 tcaatgtcga gacccataga gcccaccgca tccccagcat gcctgctatt gtcttcccaa
     8881 tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga
     8941 caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc accttccagg
     9001 gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagtc
     9061 gaggctgatc agcgggttta taacttcgta taatgtatgc tatacgaagt tatggtacct
     9121 tactcgtcca tgccgccggt ggagtggcgg ccctcggcgc gttcgtactg ttccacgatg
     9181 gtgtagtcct cgttgtggga ggtgatgtcc aacttgatgt tgacgttgta ggcgccgggc
     9241 agctgcacgg gcttcttggc cttgtaggtg gtcttgacct cagcgtcgta gtggccgccg
     9301 tccttcagct tcagcctctg cttgatctcg cccttcaggg cgccgtcctc ggggtacatc
     9361 cgctcggagg aggcctccca gcccatggtc ttcttctgca ttacggggcc gtcggagggg
     9421 aagttggtgc cgcgcagctt caccttgtag atgaactcgc cgtcctgcag ggaggagtcc
     9481 tgggtcacgg tcaccacgcc gccgtcctcg aagttcatca cgcgctccca cttgaagccc
     9541 tcggggaagg acagcttcaa gtagtcgggg atgtcggcgg ggtgcttcac gtaggccttg
     9601 gagccgtaca tgaactgagg ggacaggatg tcccaggcga agggcagggg gccacccttg
     9661 gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg
     9721 atctcgaact cgtggccgtt cacggagccc tccatgtgca ccttgaagcg catgaactcc
     9781 ttgatgatgg ccatgttatc ctcctcgccc ttgctcacca tggtggcgac cggtagcgct
     9841 agcgtaggcg ccggtcacag cttggatctg taacggcgca gaacagaaaa cgaaacaaag
     9901 acgtagagtt gagcaagcag ggtcaggcaa agcgtggaga gccggctgag tctaggtagg
     9961 ctccaaggga gcgccggaca aaggcccggt ctcgacctga gctttaaact tacctagacg
    10021 gcggacgcag ttcaggaggc accacaggcg ggaggcggca gaacgcgact caaccggcgt
    10081 ggatggcggc ctcaggtagg gcggcgggcg cgtgaaggag agatgcgagc ccctcgaagc
    10141 ttcagctgtg ttctggcggc aaacccgttg cgaaaaagaa cgttcacggc gactactgca
    10201 cttatatacg gttctccccc accctcggga aaaaggcgga gccagtacac gacatcactt
    10261 tcccagttta ccccgcgcca ccttctctag gcaccggttc aattgccgac ccctcccccc
    10321 aacttctcgg ggactgtggg cgatgtgcgc tctgcccact gacgggcacc ggagcgatcg
    10381 cagatcatgc atccatagag cccaccgcat ccccagcatg cctgctattg tcttcccaat
    10441 cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac
    10501 aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg
    10561 tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacaggtt
    10621 taaacgggcc ctctagatca ggcaccgggc ttgcgggtca tgcaccaggt gcgcggtcct
    10681 tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc gctcgtagaa ggggaggttg
    10741 cggggcgcgg aggtctccag gaaggcgggc accccggcgc gctcggccgc ctccactccg
    10801 gggagcacga cggcgctgcc cagacccttg ccctggtggt cgggcgagac gccgacggtg
    10861 gccaggaacc acgcgggctc cttgggccgg tgcggcgcca ggaggccttc catctgttgc
    10921 tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg ggccgatctc ggcgaacacc
    10981 gcccccgctt cgacgctctc cggcgtggtc cagaccgcca ccgcggcgcc gtcgtccgcg
    11041 acccacacct tgccgatgtc gagcccgacg cgcgtgagga agagttcttg cagctcggtg
    11101 acccgctcga tgtggcggtc cggatcgacg gtgtggcgcg tggcggggta gtcggcgaac
    11161 gcggcggcga gggtgcgtac ggccctgggg acgtcgtcgc gggtggcgag gcgcaccgtg
    11221 ggcttgtact cggtcatctc gagcctaggg ccgggattct cctccacgtc accgcatgtt
    11281 agaagacttc ctctgccctc tccgctgcca gatctctcga ggccctgtgg gaggaagaga
    11341 agaggtcaga agcttataac ttcgtataat gtatgctata cgaagttatt gccccactgt
    11401 ggggtggagg ggacagataa aagtacccag aaccagagcc acattaaccg gccctgggaa
    11461 tataaggtgg tcccagctcg gggacacagg atccctggag gcagcaaaca tgctgtcctg
    11521 aagtggacat aggggcccgg gttggaggaa gaagactagc tgagctctcg gacccctgga
    11581 agatgccatg acagggggct ggaagagcta gcacagacta gagaggtaag gggggtaggg
    11641 gagctgccca aatgaaagga gtgagaggtg acccgaatcc acaggagaac ggggtgtcca
    11701 ggcaaagaaa gcaagaggat ggagaggtgg ctaaagccag ggagacgggg tactttgggg
    11761 ttgtccagaa aaacggtgat gatgcaggcc tacaagaagg ggaggcggga cgcaagggag
    11821 acatccgtcg gagaaggcca tcctaagaaa cgagagatgg cacaggcccc agaaggagaa
    11881 ggaaaaggga acccagcgag tgaagacggc atggggttgg gtgagggagg agagatgccc
    11941 ggagaggacc cagacacggg gaggatccgc tcagaggaca tcacgtggtg cagcgccgag
    12001 aaggaagtgc tccggaaaga gcatccttgg gcagcaacac agcagagagc aaggggaaga
    12061 gggagtggag gaagacggaa cctgaaggag gcggcaggga aggatctggg ccagccgtag
    12121 aggtgaccca ggccacaagc tgcagacaga aagcggcaca ggcccagggg agagaatgct
    12181 ggtcagagaa agcaaagggc gaattcaacc ctgataaatg cttcaataat attgaaaaag
    12241 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
    12301 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
    12361 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
    12421 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
    12481 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
    12541 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
    12601 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
    12661 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
    12721 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
    12781 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
    12841 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
    12901 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
    12961 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
    13021 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
    13081 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
    13141 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
    13201 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
    13261 aaagatcaaa ggatcttc
//
