LOCUS       sequence-482886-       12897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAVS1 safe-harbor integration of dox-inducible NFIA-SOX9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12897)
  AUTHORS   Leng K, Rose IVL, Kim H, Xia W, Romero-Fernandez W, Rooney B, Koontz
            M, Li E, Ao Y, Wang S, Krawczyk M, Tcw J, Goate A, Zhang Y, Ullian 
            EM, Sofroniew MV, Fancy SPJ, Schrag MS, Lippmann ES, Kampmann M
  TITLE     CRISPRi screens in human iPSC-derived astrocytes elucidate 
            regulators of distinct inflammatory reactive states.
  JOURNAL   Nat Neurosci. 2022 Nov;25(11):1528-1542. doi: 
            10.1038/s41593-022-01180-9. Epub 2022 Oct 27.
  PUBMED    36303069
REFERENCE   2  (bases 1 to 12897)
  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: 
            "10.1038/s41593-022-01180-9"; journalName: "Nat Neurosci"; date: 
            "2022-11"; volume: "25"; issue: "11"; pages: "1528-1542"
COMMENT     Sequence Label: pKL100
FEATURES             Location/Qualifiers
     source          1..12897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..659
                     /label=chicken beta-actin promoter
     intron          660..1676
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1600..1622
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1684..1703
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1735..2481
                     /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(2572..2591)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2637..2692
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2691..2710)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     3002..3019
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3136..3157
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3172..3177,3178..3195,3196..3202)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3210..3226
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3215..3237
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3234..3250
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3234..3250
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3299..3420
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3310..3329)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3364..3383
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4967..5020)
                     /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"
     primer_bind     complement(6639..6663)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    6708..6726
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6744..6762
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6780..6798
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6816..6834
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6852..6870
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6888..6906
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6924..6942
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    6975..7811
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     rep_origin      complement(7859..8447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7939..7958)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8618..9409,9410..9478))
                     /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     9241..9260
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_feature    9532..10335
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     protein_bind    complement(10337..10370)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    10377..10402
                     /label=SA
                     /note="splice acceptor site"
     CDS             10426..10479
                     /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             10489..11088
                     /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(10489..10508)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     10985..11005
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    11109..11333
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        11369..11580
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             11593..11861
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      11899..11908
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(12050..12068)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     12318..12337
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(12613..12646)
                     /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(12662..12679)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12668..12892
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg
      541 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      961 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
     1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
     1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
     1201 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
     1261 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
     1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
     1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     1621 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     1681 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaatgt cgacatgagc
     1741 cggctggaca agagcaaagt gatcaacagc gccctggaac tgctgaacgg cgtgggcatc
     1801 gagggcctga ccacccggaa gctggcccag aaactgggcg tggaacagcc caccctgtac
     1861 tggcacgtga agaacaagcg ggccctgctg gacgccctgc ccatcgagat gctggaccgg
     1921 caccacaccc acagctgccc tctggaaggc gagagctggc aggacttcct gcggaacaac
     1981 gccaagagct acagatgcgc cctgctgagc caccgggacg gcgccaaagt gcacctgggc
     2041 accagaccca ccgagaagca gtacgagaca ctggaaaacc agctggcctt cctgtgccag
     2101 cagggcttca gcctggaaaa cgccctgtac gccctgagcg ccgtgggcca cttcaccctg
     2161 ggctgcgtgc tggaagaaca ggaacaccag gtcgccaaag aggaaagaga gacacccacc
     2221 accgacagca tgccccccct gctgaagcag gccatcgagc tgttcgacag acagggcgcc
     2281 gagcccgcct tcctgttcgg cctggaactg atcatctgcg gcctcgagaa gcagctgaag
     2341 tgcgagagcg gcggacccac cgacgccctg gacgacttcg acctggacat gctgcccgcc
     2401 gatgccctgg atgattttga tctggatatg ctccctgctg acgctctcga tgacttcgat
     2461 ctcgatatgc tgccaggcta aaacgcgtga attcactcct caggtgcagg ctgcctatca
     2521 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc
     2581 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata
     2641 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     2701 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc
     2761 aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta
     2821 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta
     2881 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt
     2941 acatgtgcgg ccgtcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3001 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     3061 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     3121 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     3181 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     3241 cagctatgac catgattacg ccaagctcta gctagaggtc gacggtatac agacatgata
     3301 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     3361 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     3421 ggggtgggcg atcgattcaa ggtcgagtga gctgtgtgta gacgggttgt tcccagtgct
     3481 gggggctgtg ggtctgcggg atggaaggga ccccagaggt gtcggcgatg ggggtgtaca
     3541 tggggcgctg agcggggttc atgtaggtga aggtggagta gaggccggtg ccctggcctg
     3601 ccgcgtggct gtagtaggag ctggagttct ggtggtcggt gtagtcgtac tgtgagcggg
     3661 tgatgggcgg gtaggagggg ctgtagtgtg ggaggttgaa ggggctgtag gcgatctgtt
     3721 ggggcgagtg ctgctgctgc tcgctgtagt ggctggggct cagctgctcc gtcttgatgt
     3781 gcgttcgctg ggactggccc ggctcgctgc tcagcgtggt cagcgtgtgc gcctgtggct
     3841 gctgcggggg tgccgccggc tgctgtgggg gcgccgcctg cggctgcggg ggcgcctgcg
     3901 gggccggcgg ggcctgtggg ggctgctgcg ggggtggcgg cggcgcctgc tgcttggaca
     3961 tccacacgtg gcccgcgctc gccggggtgg ccgcggtgct gctgatgccg tagctgcccg
     4021 tgtaggtgac ctggccgtgc gtggccggca cccccgggtg gccgttgggc ggcaggtact
     4081 ggtcaaactc gttgacatcg aaggtctcga tgttggagat gacgtcgctg ctcagctcgc
     4141 cgatgtccac gtcgcggaag tcgatagggg gctgtctgcc cccctctggc aaggggcgcc
     4201 cctctcgctt caggtcagcc ttgcccggct gcacgtcggt tttgggggtg gtgggtgggg
     4261 tcggtgggcc ctgggattgc cccgagtgct cgccggggga gtgcacctcg ctcatgccgg
     4321 aggaggagtg tggcgagtcg gcctgcagcg ccttgaagat ggcgttgggg gagatgtgcg
     4381 tctgctccgt ggcctcctct gcctccgcct gcccgttctt caccgacttc ctccgccgcg
     4441 gctggtactt gtaatccggg tggtccttct tgtgctgcac gcgcagccgc tccgcctcct
     4501 ccacgaaggg ccgcttctcg ctctcgttca gaagtctcca gagcttgccc agcgtcttgc
     4561 tgagctcggc gttgtgcaag tgcgggtact ggtccgcgag cttcctgcgc gccgcctgcg
     4621 cccacaccat gaaggcgttc atgggccgct tgacgtgcgg cttgttcttg ctggagccgt
     4681 tgacgcgcac cggcatgggc accagcgtcc agtcgtagcc tttgagcacc tggctgaccg
     4741 cctcgcggat gcacacgggg aacttgtcct cctcgctctc cttcttcaga tcgggctcgc
     4801 ccttggggaa cgtgttctcc tggggccgcg tgttctcggt gtccgagccg gagcccgacg
     4861 ggcagggcga gcccgcggag tcctcggaca tggtggggct gggggcgccg gacaggccct
     4921 tctcctgctc gtcggtcatc ttcatgaagg ggtccaggag attcataggg ccgggattct
     4981 cttccacgtc accgcatgtc agcagactgc ctctgccctc tccgctacct cccaggtacc
     5041 aggactgtgt ctgttgaggg atatttacaa agcttggatc ccgtggtcca acactgacga
     5101 atcggtttgc gggggaggtg ctgggtgtcg agtaggttgg tgacgtgggg gaggctgcac
     5161 ctccttctgt tgacgtggtt ggaggctttg tgtctggcac cggcagaggc acaggcctgg
     5221 ccatcggtgg tggcggtggc ggtggcaaca ttggggtggg aaggaatggg ttgtgcacct
     5281 tgccttggct gctcccattg ggattgagga accccacctg tccagcctgc tgaccagcat
     5341 cagggcagac aagttggaca aattctttca gcgtctcctg agggtgatag cggatggctg
     5401 ggtgtgagaa gtaaggccca ggctgctgga taatgggtga tgtcgggaaa tgaagagtcg
     5461 atggtgttgc atgcggactt gctctgggtc ctgtaatgac aggtcggtga tgctgtgtga
     5521 acgccgttcc cagggaggag gtctgtgaag gggaggggct gctgaaacca gacttctccg
     5581 acttcttcag tgtggttgga gatggcattc ctggctccac ttcatgccat ccacttgact
     5641 gactgccact tcctggggag cgcccttggc ctgtataaaa tggctcctca ccaggactgt
     5701 ccatttcatc ctccacagac ttgaggcgct ttgtggagct cgtagaggat gtgctgggta
     5761 aagacctcct cattgctcct ggactcatgc tgtagtatga agaactttcc aaatctgaga
     5821 gagaaaaatt tgggccagtt cctgcagcta ttggtgtctg tgacactctt actagctcag
     5881 tgacactaaa aacacctgat gtgacaaaac tgtcctggaa gcccaaatgt ccattttctg
     5941 gctggtcctt aatgtcagcg tcacttggct ggctgggact ttcagattga cttgaatctg
     6001 ctgcatgcac aaagtatgcc aaatataaat cgagttcctt aacagaaacc cctatgtgat
     6061 ggggttggac acagagccct ggattagagc attgtgggga ctttacaagg cgctcgccat
     6121 cagtactttc cagcggaata cctttaaaca aaatcaccat aacaaggtcc aacctccaga
     6181 ctttatctgc ctggcggagg cagtcaattc ttcgcatctt gcctttctgg tctgggttgg
     6241 aaagaacaca acatggaggt tttttccctg taactgtaag aacaaaatcc tctcgatatt
     6301 cgggtcggat atctttccgc aactttgcca gaagtcgaga tgcccacttc tgcttgacct
     6361 ctggtttttc acttagcaat tcatccttca cggctctctc ttcttctttt gacatacgct
     6421 tttcatgttt tttgaagtat tttcgttttc gggcctgcag gttgaaccat gtgtaggcaa
     6481 aggctcggac gtggggcaga agtgcttcga tgaaaggatg aaattcatcc tgggtgagac
     6541 agagcggaga atacatggtg gcggccttaa gtttacgagg gtaggaagtg gtacggaaag
     6601 ttggtataag acaaaagtgt tgtggaattg ctccaggcga tctgacggtt cactaaacga
     6661 gctctgcttt tataggcgcc caccgtacac gcctaaagct tatacgttct ctatcactga
     6721 tagggagtaa actggatata cgttctctat cactgatagg gagtaaactg tagatacgtt
     6781 ctctatcact gatagggagt aaactggtca tacgttctct atcactgata gggagtaaac
     6841 tccttatacg ttctctatca ctgataggga gtaaagtctg catacgttct ctatcactga
     6901 tagggagtaa actcttcata cgttctctat cactgatagg gagtaaacca ctgtggggtg
     6961 gagatatcag ctttactagg gacaggattg gtgacagaaa agccccatcc ttaggcctcc
     7021 tccttcctag tctcctgata ttgggtctaa cccccacctc ctgttaggca gattccttat
     7081 ctggtgacac acccccattt cctggagcca tctctctcct tgccagaacc tctaaggttt
     7141 gcttacgatg gagccagaga ggatcctggg agggagagct tggcaggggg tgggagggaa
     7201 gggggggatg cgtgacctgc ccggttctca gtggccaccc tgcgctaccc tctcccagaa
     7261 cctgagctgc tctgacgcgg ctgtctggtg cgtttcactg atcctggtgc tgcagcttcc
     7321 ttacacttcc caagaggaga agcagtttgg aaaaacaaaa tcagaataag ttggtcctga
     7381 gttctaactt tggctcttca cctttctagt ccccaattta tattgttcct ccgtgcgtca
     7441 gttttacctg tgagataagg ccagtagcca gccccgtcct ggcagggctg tggtgaggag
     7501 gggggtgtcc gtgtggaaaa ctccctttgt gagaatggtg cgtcctaggt gttcaccagg
     7561 tcgtggccgc ctctactccc tttctctttc tccatccttc tttccttaaa gagtccccag
     7621 tgctatctgg gacatattcc tccgcccaga gcagggtccc gcttccctaa ggccctgctc
     7681 tgggcttctg ggtttgagtc cttggcaagc ccaggagagg cgctcaggct tccctgtccc
     7741 ccttcctcgt ccaccatctc atgcccctgg ctctcctgcc ccttccctac aggggttcct
     7801 ggctctgctc taagggcgaa ttcgccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7861 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     7921 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     7981 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     8041 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     8101 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     8161 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     8221 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     8281 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     8341 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     8401 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     8461 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     8521 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     8581 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     8641 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     8701 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     8761 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     8821 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     8881 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     8941 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     9001 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     9061 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     9121 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     9181 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     9241 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     9301 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     9361 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     9421 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     9481 tcttcctttt tcaatattat tgaagcattt atcagggttg aattcgccct ttgctttctc
     9541 tgaccagcat tctctcccct gggcctgtgc cgctttctgt ctgcagcttg tggcctgggt
     9601 cacctctacg gctggcccag atccttccct gccgcctcct tcaggttccg tcttcctcca
     9661 ctccctcttc cccttgctct ctgctgtgtt gctgcccaag gatgctcttt ccggagcact
     9721 tccttctcgg cgctgcacca cgtgatgtcc tctgagcgga tcctccccgt gtctgggtcc
     9781 tctccgggca tctctcctcc ctcacccaac cccatgccgt cttcactcgc tgggttccct
     9841 tttccttctc cttctggggc ctgtgccatc tctcgtttct taggatggcc ttctccgacg
     9901 gatgtctccc ttgcgtcccg cctccccttc ttgtaggcct gcatcatcac cgtttttctg
     9961 gacaacccca aagtaccccg tctccctggc tttagccacc tctccatcct cttgctttct
    10021 ttgcctggac accccgttct cctgtggatt cgggtcacct ctcactcctt tcatttgggc
    10081 agctccccta ccccccttac ctctctagtc tgtgctagct cttccagccc cctgtcatgg
    10141 catcttccag gggtccgaga gctcagctag tcttcttcct ccaacccggg cccctatgtc
    10201 cacttcagga cagcatgttt gctgcctcca gggatcctgt gtccccgagc tgggaccacc
    10261 ttatattccc agggccggtt aatgtggctc tggttctggg tacttttatc tgtcccctcc
    10321 accccacagt ggggcaataa cttcgtatag catacattat acgaagttat aagcttctga
    10381 cctcttctct tcctcccaca gggcctcgag agatctggca gcggagaggg cagaggaagt
    10441 cttctaacat gcggtgacgt ggaggagaat cccggcccta ggctcgagat gaccgagtac
    10501 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
    10561 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
    10621 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
    10681 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
    10741 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
    10801 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
    10861 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
    10921 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
    10981 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
    11041 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgatc tagagggccc
    11101 gtttaaacct gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
    11161 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
    11221 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
    11281 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggatgcatg
    11341 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
    11401 aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac
    11461 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
    11521 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
    11581 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc
    11641 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg
    11701 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct
    11761 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc
    11821 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct
    11881 acgctagcgc taccggtcgc caccatggtg agcaagggcg aggaggataa catggccatc
    11941 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
    12001 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
    12061 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
    12121 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
    12181 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
    12241 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
    12301 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
    12361 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
    12421 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
    12481 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
    12541 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
    12601 gagtaaggta ccataacttc gtatagcata cattatacga agttataaac ccgctgatca
    12661 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
    12721 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
    12781 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
    12841 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat gggtctc
//
