LOCUS       sequence-216317-       12553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DR-GFP substrate (Addgene #26475) flanked by human AAVS1 homology 
            arms. Endogenous AAVS1 promoter drives T2A-hygromycin expression 
            after integration. DR-GFP cassette contains PGK-puromycin. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12553)
  TITLE     CRISPR-mediated insertion of DR-GFP into the AAVS1 locus
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 12553)
  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: 
            "Unpublished"
COMMENT     Sequence Label: AAVS1-DRGFP
FEATURES             Location/Qualifiers
     source          1..12553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..43)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(23..41)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(62..78)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    86..102
                     /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(110..116,117..134,135..140))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    155..176
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(293..310)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(464..1052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(544..563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1223..2014,2015..2083))
                     /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     1846..1865
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2084..2188)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2214..2669)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2351..2372)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2563..2582
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2796..2818
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2810..2827
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2811..2827
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2837..2856
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2837..2855
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2878..3681
                     /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    3688..3713
                     /label=SA
                     /note="splice acceptor site"
     CDS             3737..3790
                     /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             3800..4825
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3800..4825
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     complement(4857..4874)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4863..5087
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    5094..5175
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        5195..5574
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5577..5852
                     /label=chicken beta-actin promoter
     intron          5853..6861
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     6785..6807
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     6869..6888
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             6946..6966
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6988..7008
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      7222..7231
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7228..7230,7231..7233,7234..7947)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSG*G*QGNTYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     7881..7902
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    8070..8191
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     8161..8180
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(8852..9451)
                     /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(8935..8955)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     9432..9451
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(9470..9967)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(9518..9537)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     9909..9932
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     complement(10055..10074)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    10120..10175
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(10174..10193)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     CDS             10537..10557
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10579..10599
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      10813..10822
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(10819..10821,10822..10824,10825..11283)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMA"
     primer_bind     complement(11340..11356)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    11364..11380
                     /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(11388..11394,11395..11412,11413..11418))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11433..11454
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      11559..11694
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    join(11721..12553,1..4)
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
ORIGIN
        1 ctctggtacc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
       61 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
      121 ccgggagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
      181 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
      241 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
      301 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
      361 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
      421 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
      481 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
      541 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
      601 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
      661 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
      721 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
      781 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
      841 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
      901 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
      961 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     1021 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     1081 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     1141 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     1201 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     1261 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     1321 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     1381 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     1441 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     1501 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     1561 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     1621 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     1681 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     1741 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     1801 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     1861 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     1921 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     1981 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     2041 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     2101 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     2161 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     2221 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2281 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     2341 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     2401 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     2461 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     2521 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     2581 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     2641 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
     2701 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     2761 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     2821 ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttggagctcc accgcggtgc
     2881 tttctctgac cagcattctc tcccctgggc ctgtgccgct ttctgtctgc agcttgtggc
     2941 ctgggtcacc tctacggctg gcccagatcc ttccctgccg cctccttcag gttccgtctt
     3001 cctccactcc ctcttcccct tgctctctgc tgtgttgctg cccaaggatg ctctttccgg
     3061 agcacttcct tctcggcgct gcaccacgtg atgtcctctg agcggatcct ccccgtgtct
     3121 gggtcctctc cgggcatctc tcctccctca cccaacccca tgccgtcttc actcgctggg
     3181 ttcccttttc cttctccttc tggggcctgt gccatctctc gtttcttagg atggccttct
     3241 ccgacggatg tctcccttgc gtcccgcctc cccttcttgt aggcctgcat catcaccgtt
     3301 tttctggaca accccaaagt accccgtctc cctggcttta gccacctctc catcctcttg
     3361 ctttctttgc ctggacaccc cgttctcctg tggattcggg tcacctctca ctcctttcat
     3421 ttgggcagct cccctacccc ccttacctct ctagtctgtg ctagctcttc cagccccctg
     3481 tcatggcatc ttccaggggt ccgagagctc agctagtctt cttcctccaa cccgggcccc
     3541 tatgtccact tcaggacagc atgtttgctg cctccaggga tcctgtgtcc ccgagctggg
     3601 accaccttat attcccaggg ccggttaatg tggctctggt tctgggtact tttatctgtc
     3661 ccctccaccc cacagtgggg cagatccctg acctcttctc ttcctcccac agggcctcga
     3721 gagatctggc agcggagagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa
     3781 tcccggccct aggctgcaga tgaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt
     3841 tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc
     3901 tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc
     3961 cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat
     4021 tccggaagtg cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg
     4081 tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc
     4141 ggtcgcggag gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg
     4201 cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat
     4261 tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt
     4321 cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct
     4381 cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt
     4441 cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt
     4501 ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc
     4561 ggagcttgca ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact
     4621 ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga
     4681 cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc
     4741 ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag
     4801 cactcgtccg agggcaaagg aatagtctag agggcccgtt taaacccgct gatcagcctc
     4861 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     4921 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     4981 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     5041 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatgggtc gaggcagtga
     5101 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     5161 tgcaataaac aagttaacaa caacaattct ggtcgacatt gattattgac tagttattaa
     5221 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     5281 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     5341 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     5401 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc
     5461 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     5521 cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catgggtcga
     5581 ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt
     5641 gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggcgcg
     5701 cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg
     5761 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc
     5821 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgttgcct tcgccccgtg
     5881 ccccgctccg cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac
     5941 aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac
     6001 ggctcgtttc ttttctgtgg ctgcgtgaaa gccttaaagg gctccgggag ggccctttgt
     6061 gcggggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc
     6121 ggcccgcgct gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc
     6181 gcgtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg gggctgcgag
     6241 gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg
     6301 gcggtcgggc tgtaaccccc ccctgcaccc ccctccccga gttgctgagc acggcccggc
     6361 ttcgggtgcg gggctccgtg cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg
     6421 gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga
     6481 ggggcgcggc ggccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc
     6541 cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctggcggagc
     6601 cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cgggcgaagc ggtgcggcgc
     6661 cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct
     6721 ccatctccag cctcggggct gccgcagggg gacggctgcc ttcggggggg acggggcagg
     6781 gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat
     6841 gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat
     6901 tttggcaaag aattcagatc cgccgccact atgggatcaa gatcgccaaa aaagaagaga
     6961 aaggtgccga agaagcatgc agcaccacca aaaaaaaaac gaaaagtaga agacccacga
     7021 ggcaacacca gcggcgtgct gagcaccccc aaggccaaga gggccaagca cccccccggc
     7081 accgagaagc ccaggagcag gagccagagc gagcagcccg ccacctgccc catctgctac
     7141 gccgtgatca ggcagagcag gaacctgagg aggcacctgg agctgaggca cttcgccaag
     7201 cccggcgtgg atccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     7261 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     7321 ggctagggat aacagggtaa tacctacggc aagctgaccc tgaagttcat ctgcaccacc
     7381 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     7441 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     7501 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     7561 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     7621 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     7681 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     7741 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     7801 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     7861 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     7921 ctcggcatgg acgagctgta caagtaaagc ggccgcgact ctagatcata atcagccata
     7981 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     8041 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     8101 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     8161 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatctgaag
     8221 cttctgatgg aattagaact tggcaaaaca atactgagaa tgaagtgtat gtggaacaga
     8281 ggctgctgat ctcgttcttc aggctatgaa actgacacat ttggaaacca cagtacttag
     8341 aaccacaaag tgggaatcaa gagaaaaaca atgatcccac gagagatcta tagatctata
     8401 gatcatgagt gggaggaatg agctggccct taatttggtt ttgcttgttt aaattatgat
     8461 atccaactat gaaacattat cataaagcaa tagtaaagag ccttcagtaa agagcaggca
     8521 tttatctaat cccaccccac ccccaccccc gtagctccaa tccttccatt caaaatgtag
     8581 gtactctgtt ctcacccttc ttaacaaagt atgacaggaa aaacttccat tttagtggac
     8641 atctttattg tttaatagat catcaatttc gatccgctcc tgggcaccga actgcgccgc
     8701 gtgttcagca gggtcggcgt gttcggtgtg tcccccgcgg tgggcctcgg gggcgggtgc
     8761 ggggtcggcg gggccgcccc gggtggcttc ggtcggagcc atggggtcgt gcgctccttt
     8821 cggtcgggcg ctgcgggtcg tggggcgggc gtcaggcacc gggcttgcgg gtcatgcacc
     8881 aggtgcgcgg tccttcgggc acctcgacgt cggcggtgac ggtgaagccg agccgctcgt
     8941 agaaggggag gttgcggggc gcggaggtct ccaggaaggc gggcaccccg gcgcgctcgg
     9001 ccgcctccac tccggggagc acgacggcgc tgcccagacc cttgccctgg tggtcgggcg
     9061 agacgccgac ggtggccagg aaccacgcgg gctccttggg ccggtgcggc gccaggaggc
     9121 cttccatctg ttgctgcgcg gccagccggg aaccgctcaa ctcggccatg cgcgggccga
     9181 tctcggcgaa caccgccccc gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg
     9241 cgccgtcgtc cgcgacccac accttgccga tgtcgagccc gacgcgcgtg aggaagagtt
     9301 cttgcagctc ggtgacccgc tcgatgtggc ggtccgggtc gacggtgtgg cgcgtggcgg
     9361 ggtagtcggc gaacgcggcg gcgagggtgc gtacggcccg ggggacgtcg tcgcgggtgg
     9421 cgaggcgcac cgtgggcttg tactcggtca tggtggcggc tggatcggtc gaaaggcccg
     9481 gagatgagga agaggagaac agcgcggcag acgtgcgctt ttgaagcgtg cagaatgccg
     9541 ggcctccgga ggaccttcgg gcgcccgccc cgcccctgag cccgcccctg agcccgcccc
     9601 cggacccacc ccttcccagc ctctgagccc agaaagcgaa ggagcaaagc tgctattggc
     9661 cgctgcccca aaggcctacc cgcttccatt gctcagcggt gctgtccatc tgcacgagac
     9721 tagtgagacg tgctacttcc atttgtcacg tcctgcacga cgcgagctgc ggggcggggg
     9781 ggaacttcct gactagggga ggagtagaag gtggcgcgaa ggggccacca aagaacggag
     9841 ccggttggcg cctaccggtg gatgtggaat gtgtgaggcc agaggccact tgtgtagcgc
     9901 caagtgccca gcggggctgc taaagcgcat gctccagact gccttgggaa aagcgcctcc
     9961 cctacccggt agaattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg
    10021 tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg
    10081 gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca
    10141 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
    10201 atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat gccatatgct
    10261 ggctgccatg aacaaaggtg gctataaaga ggtcatcagt atatgaaaca gccccctgct
    10321 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
    10381 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
    10441 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atgaagatcc
    10501 ctcgacctgc agcccaagct ttagggatca agatcgccaa aaaagaagag aaaggtgccg
    10561 aagaagcatg cagcaccacc aaaaaaaaaa cgaaaagtag aagacccacg aggcaacacc
    10621 agcggcgtgc tgagcacccc caaggccaag agggccaagc acccccccgg caccgagaag
    10681 cccaggagca ggagccagag cgagcagccc gccacctgcc ccatctgcta cgccgtgatc
    10741 aggcagagca ggaacctgag gaggcacctg gagctgaggc acttcgccaa gcccggcgtg
    10801 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
    10861 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
    10921 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
    10981 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
    11041 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
    11101 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
    11161 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
    11221 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
    11281 gccgacaagc agaagaacgg catcaaggtg aacttcaata aaagcttggc gtaatcatgg
    11341 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
    11401 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
    11461 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
    11521 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
    11581 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
    11641 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagtac
    11701 catggaatgc atccctcgag actagggaca ggattggtga cagaaaagcc ccatccttag
    11761 gcctcctcct tcctagtctc ctgatattgg gtctaacccc cacctcctgt taggcagatt
    11821 ccttatctgg tgacacaccc ccatttcctg gagccatctc tctccttgcc agaacctcta
    11881 aggtttgctt acgatggagc cagagaggat cctgggaggg agagcttggc agggggtggg
    11941 agggaagggg gggatgcgtg acctgcccgg ttctcagtgg ccaccctgcg ctaccctctc
    12001 ccagaacctg agctgctctg acgcggctgt ctggtgcgtt tcactgatcc tggtgctgca
    12061 gcttccttac acttcccaag aggagaagca gtttggaaaa acaaaatcag aataagttgg
    12121 tcctgagttc taactttggc tcttcacctt tctagtcccc aatttatatt gttcctccgt
    12181 gcgtcagttt tacctgtgag ataaggccag tagccagccc cgtcctggca gggctgtggt
    12241 gaggaggggg gtgtccgtgt ggaaaactcc ctttgtgaga atggtgcgtc ctaggtgttc
    12301 accaggtcgt ggccgcctct actccctttc tctttctcca tccttctttc cttaaagagt
    12361 ccccagtgct atctgggaca tattcctccg cccagagcag ggtcccgctt ccctaaggcc
    12421 ctgctctggg cttctgggtt tgagtccttg gcaagcccag gagaggcgct caggcttccc
    12481 tgtccccctt cctcgtccac catctcatgc ccctggctct cctgcccctt ccctacaggg
    12541 gttcctggct ctg
//
