LOCUS       sequence-319218-       13987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAVS1 targeting donor plasmid with Bi-DREADD expression cassette 
            (hM3Dq-mCherry-P2A-HA-KORD) and puromycin selection gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13987)
  AUTHORS   Xiong M, Tao Y, Gao Q, Feng B, Yan W, Zhou Y, Kotsonis TA, Yuan T, 
            You Z, Wu Z, Xi J, Haberman A, Graham J, Block J, Zhou W, Chen Y, 
            Zhang SC
  TITLE     Human Stem Cell-Derived Neurons Repair Circuits and Restore Neural 
            Function.
  JOURNAL   Cell Stem Cell. 2020 Sep 16. pii: S1934-5909(20)30410-0. doi: 
            10.1016/j.stem.2020.08.014.
  PUBMED    32966778
REFERENCE   2  (bases 1 to 13987)
  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"; journalName: "Cell Stem 
            Cell. 2020 Sep 16. pii: S1934-5909(20)30410-0. doi: 
            10.1016/j.stem.2020.08.014."
COMMENT     Sequence Label: AAVS1-pur-CAG-Bi-DREADD
FEATURES             Location/Qualifiers
     source          1..13987
                     /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"
     regulatory      3531..3540
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3537..4244
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3682..3700)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3950..3969
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4260..4316
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4320..4346
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(4980..4991)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     misc_feature    5523..6111
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5576..5596)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5994..6005)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6143..6619
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(6342..6361)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(6729..6748)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6794..6849
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6848..6867)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7210..7226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7234..7250
                     /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(7258..7264,7265..7282,7283..7288))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7303..7324
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    7398..8234
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(8268..8287)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8269..8287)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8294..8311)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8294..8310)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8303..8325)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             8448..8750
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     8629..8648
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(9153..9172)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9767..9786
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(10147..10938,10939..11007))
                     /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     10770..10789
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11131..11719
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11620..11639
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11873..11890
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12007..12028
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12043..12048,12049..12066,12067..12073)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12081..12097
                     /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     12105..12121
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        12142..12160
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    12199..13002
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     misc_feature    13009..13034
                     /label=SA
                     /note="splice acceptor site"
     CDS             13058..13111
                     /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             13121..13720
                     /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(13121..13140)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     13617..13637
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(13752..13769)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    13758..13982
                     /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 crggsggggs ggggsggggs grggggsggg
      541 gsggggcgag 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 ggccggggrg 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 caaagaattc gtcgacgccg
     1741 ccaccatgac cttgcacaat aacagtacaa cctcgccttt gtttccaaac atcagctcct
     1801 cctggataca cagcccctcc gatgcagggc tgcccccggg aaccgtcact catttcggca
     1861 gctacaatgt ttctcgagca gctggcaatt tctcctctcc agacggtacc accgatgacc
     1921 ctctgggagg tcataccgtc tggcaagtgg tcttcatcgc tttcttaacg ggcatcctgg
     1981 ccttggtgac catcatcggc aacatcctgg taattgtgtc atttaaggtc aacaagcagc
     2041 tgaagacggt caacaactac ttcctcttaa gcctggcctg tgccgatctg attatcgggg
     2101 tcatttcaat gaatctgttt acgacctaca tcatcatgaa tcgatgggcc ttagggaact
     2161 tggcctgtga cctctggctt gccattgact gcgtagccag caatgcctct gttatgaatc
     2221 ttctggtcat cagctttgac agatactttt ccatcacgag gccgctcacg taccgagcca
     2281 aacgaacaac aaagagagcc ggtgtgatga tcggtctggc ttgggtcatc tcctttgtcc
     2341 tttgggctcc tgccatcttg ttctggcaat actttgttgg aaagagaact gtgcctccgg
     2401 gagagtgctt cattcagttc ctcagtgagc ccaccattac ttttggcaca gccatcgctg
     2461 gtttttatat gcctgtcacc attatgacta ttttatactg gaggatctat aaggaaactg
     2521 aaaagcgtac caaagagctt gctggcctgc aagcctctgg gacagaggca gagacagaaa
     2581 actttgtcca ccccacgggc agttctcgaa gctgcagcag ttacgaactt caacagcaaa
     2641 gcatgaaacg ctccaacagg aggaagtatg gccgctgcca cttctggttc acaaccaaga
     2701 gctggaaacc cagctccgag cagatggacc aagaccacag cagcagtgac agttggaaca
     2761 acaatgatgc tgctgcctcc ctggagaact ccgcctcctc cgacgaggag gacattggct
     2821 ccgagacgag agccatctac tccatcgtgc tcaagcttcc gggtcacagc accatcctca
     2881 actccaccaa gttaccctca tcggacaacc tgcaggtgcc tgaggaggag ctggggatgg
     2941 tggacttgga gaggaaagcc gacaagctgc aggcccagaa gagcgtggac gatggaggca
     3001 gttttccaaa aagcttctcc aagcttccca tccagctaga gtcagccgtg gacacagcta
     3061 agacttctga cgtcaactcc tcagtgggta agagcacggc cactctacct ctgtccttca
     3121 aggaagccac tctggccaag aggtttgctc tgaagaccag aagtcagatc actaagcgga
     3181 aaaggatgtc cctggtcaag gagaagaaag cggcccagac cctcagtgcg atcttgcttg
     3241 ccttcatcat cacttggacc ccatacaaca tcatggttct ggtgaacacc ttttgtgaca
     3301 gctgcatacc caaaaccttt tggaatctgg gctactggct gtgctacatc aacagcaccg
     3361 tgaaccccgt gtgctatgct ctgtgcaaca aaacattcag aaccactttc aagatgctgc
     3421 tgctgtgcca gtgtgacaaa aaaaagaggc gcaagcagca gtaccagcag agacagtcgg
     3481 tcatttttca caagcgcgca cccgagcagg ccttgaagga tcccccggtc gccaccatgg
     3541 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     3601 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     3661 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     3721 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     3781 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     3841 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     3901 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     3961 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     4021 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     4081 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     4141 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     4201 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtccgga ggatccggcg
     4261 caacaaactt ctctctgctg aaacaagccg gagatgtcga agagaatcct ggaccgatgt
     4321 acccatacga tgttccagat tacgctatgg aatccccgat tcagatcttc cgcggggagc
     4381 cgggccctac ctgcgccccg agcgcctgcc tgccccccaa cagcagcgcc tggtttcccg
     4441 gctgggccga gcccgacagc aacggcagcg ccggctcgga ggacgcgcag ctggagcccg
     4501 cgcacatctc cccggccatc ccggtcatca tcacggcggt ctactccgta gtgttcgtcg
     4561 tgggcttggt gggcaactcg ctggtcatgt tcgtgatcat ccgatacaca aagatgaaga
     4621 cagcaaccaa catttacata tttaacctgg ctttggcaga tgctttagtt actacaacca
     4681 tgccctttca gagtacggtc tacttgatga attcctggcc ttttggggat gtgctgtgca
     4741 agatagtaat ttccattaat tactacaaca tgttcaccag catcttcacc ttgaccatga
     4801 tgagcgtgga ccgctacatt gccgtgtgcc accccgtgaa ggctttggac ttccgcacac
     4861 ccttgaaggc aaagatcatc aatatctgca tctggctgct gtcgtcatct gttggcatct
     4921 ctgcaatagt ccttggaggc accaaagtca gggaagacgt cgatgtcatt gagtgctcct
     4981 tgcagttccc agatgatgac tactcctggt gggacctctt catgaagatc tgcgtcttca
     5041 tctttgcctt cgtgatccct gtcctcatca tcatcgtctg ctacaccctg atgatcctgc
     5101 gtctcaagag cgtccggctc ctttctggct cccgagagaa agatcgcaac ctgcgtagga
     5161 tcaccagact ggtcctggtg gtggtggcag tcttcgtcgt ctgctggact cccattcaca
     5221 tattcatcct ggtggaggct ctggggagca cctcccacag cacagctgct ctctccagct
     5281 attacttctg catcgcctta ggctatacca acagtagcct gaatcccatt ctctacgcct
     5341 ttcttgatga aaacttcaag cggtgtttcc gggacttctg ctttccactg aagatgagga
     5401 tggagcggca gagcactagc agagtccgaa atacagttca ggatcctgct tacctgaggg
     5461 acatcgatgg gatgaataaa ccagtatgat agacgcgtga attcgatatc aagcttatcg
     5521 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     5581 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     5641 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     5701 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     5761 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     5821 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     5881 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     5941 tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     6001 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     6061 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc
     6121 gagcgctgct cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct
     6181 ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat
     6241 cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag
     6301 caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg
     6361 gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct
     6421 cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt
     6481 tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta
     6541 atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac
     6601 tgctcccttc cctgtccttc tgattttgta ggtaaccagg cgcgtgaatt cactcctcag
     6661 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
     6721 cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     6781 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     6841 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     6901 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta
     6961 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     7021 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     7081 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     7141 ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc
     7201 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     7261 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     7321 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat
     7381 cgacagtact aagctttact agggacagga ttggtgacag aaaagcccca tccttaggcc
     7441 tcctccttcc tagtctcctg atattgggtc taacccccac ctcctgttag gcagattcct
     7501 tatctggtga cacaccccca tttcctggag ccatctctct ccttgccaga acctctaagg
     7561 tttgcttacg atggagccag agaggatcct gggagggaga gcttggcagg gggtgggagg
     7621 gaaggggggg atgcgtgacc tgcccggttc tcagtggcca ccctgcgcta ccctctccca
     7681 gaacctgagc tgctctgacg cggctgtctg gtgcgtttca ctgatcctgg tgctgcagct
     7741 tccttacact tcccaagagg agaagcagtt tggaaaaaca aaatcagaat aagttggtcc
     7801 tgagttctaa ctttggctct tcacctttct agtccccaat ttatattgtt cctccgtgcg
     7861 tcagttttac ctgtgagata aggccagtag ccagccccgt cctggcaggg ctgtggtgag
     7921 gaggggggtg tccgtgtgga aaactccctt tgtgagaatg gtgcgtccta ggtgttcacc
     7981 aggtcgtggc cgcctctact ccctttctct ttctccatcc ttctttcctt aaagagtccc
     8041 cagtgctatc tgggacatat tcctccgccc agagcagggt cccgcttccc taaggccctg
     8101 ctctgggctt ctgggtttga gtccttggca agcccaggag aggcgctcag gcttccctgt
     8161 cccccttcct cgtccaccat ctcatgcccc tggctctcct gccccttccc tacaggggtt
     8221 cctggctctg ctctaagggc gaattcgcgg ccgctaaatt caattcgccc tatagtgagt
     8281 cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     8341 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     8401 cccgcaccga tcgcccttcc caacagttgc gcagcctata cgtacggcag tttaaggttt
     8461 acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg
     8521 acacgccggg gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag
     8581 tctcccgtga actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca
     8641 ccgatatggc cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc
     8701 gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggca
     8761 tgagattatc aaaaaggatc ttcacctaga tccttttcac gtagaaagcc agtccgcaga
     8821 aacggtgctg accccggatg aatgtcagct actgggctat ctggacaagg gaaaacgcaa
     8881 gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg gcgatagcta gactgggcgg
     8941 ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga
     9001 agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat
     9061 caagctctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     9121 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     9181 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     9241 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     9301 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     9361 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     9421 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     9481 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     9541 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     9601 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     9661 atgcatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     9721 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     9781 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     9841 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgaatt
     9901 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     9961 caccgcatca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
    10021 ctaaatacat tcaaatatgt atccgctcat gagattatca aaaaggatct tcacctagat
    10081 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
    10141 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
    10201 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
    10261 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
    10321 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
    10381 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
    10441 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
    10501 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
    10561 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
    10621 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
    10681 cttttctgtg actggtgaat actcaaccaa gtcattctga gaatagtgta tgcggcgacc
    10741 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
    10801 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
    10861 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
    10921 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
    10981 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
    11041 tcagggttat tgtctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
    11101 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
    11161 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
    11221 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
    11281 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
    11341 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
    11401 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
    11461 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
    11521 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
    11581 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
    11641 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
    11701 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
    11761 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
    11821 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
    11881 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
    11941 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
    12001 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
    12061 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
    12121 ccatgattac gccaagctca gaattaaccc tcactaaagg gactagctag tcctgcaggt
    12181 ttaaacgaat tcgccctttg ctttctctga ccagcattct ctcccctggg cctgtgccgc
    12241 tttctgtctg cagcttgtgg cctgggtcac ctctacggct ggcccagatc cttccctgcc
    12301 gcctccttca ggttccgtct tcctccactc cctcttcccc ttgctctctg ctgtgttgct
    12361 gcccaaggat gctctttccg gagcacttcc ttctcggcgc tgcaccacgt gatgtcctct
    12421 gagcggatcc tccccgtgtc tgggtcctct ccgggcatct ctcctccctc acccaacccc
    12481 atgccgtctt cactcgctgg gttccctttt ccttctcctt ctggggcctg tgccatctct
    12541 cgtttcttag gatggccttc tccgacggat gtctcccttg cgtcccgcct ccccttcttg
    12601 taggcctgca tcatcaccgt ttttctggac aaccccaaag taccccgtct ccctggcttt
    12661 agccacctct ccatcctctt gctttctttg cctggacacc ccgttctcct gtggattcgg
    12721 gtcacctctc actcctttca tttgggcagc tcccctaccc cccttacctc tctagtctgt
    12781 gctagctctt ccagccccct gtcatggcat cttccagggg tccgagagct cagctagtct
    12841 tcttcctcca acccgggccc ctatgtccac ttcaggacag catgtttgct gcctccaggg
    12901 atcctgtgtc cccgagctgg gaccacctta tattcccagg gccggttaat gtggctctgg
    12961 ttctgggtac ttttatctgt cccctccacc ccacagtggg gcaagcttct gacctcttct
    13021 cttcctccca cagggcctcg agagatctgg cagcggagag ggcagaggaa gtcttctaac
    13081 atgcggtgac gtggaggaga atcccggccc taggctcgag atgaccgagt acaagcccac
    13141 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
    13201 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
    13261 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
    13321 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
    13381 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
    13441 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
    13501 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
    13561 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
    13621 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
    13681 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga tctagagggc ccgtttaaac
    13741 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
    13801 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
    13861 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
    13921 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
    13981 gggtctc
//
