LOCUS       sequence-355459-       12573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual fluorescent reporter construct with double HS4 insulator, 
            upstream TRE (Tetracycline Response Element) recruitment site, arms 
            for integration at AAVS1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12573)
  AUTHORS   Lensch S, Herschl MH, Ludwig CH, Sinha J, Hinks MM, Mukund A, 
            Fujimori T, Bintu L
  TITLE     Dynamic spreading of chromatin-mediated gene silencing and 
            reactivation between neighboring genes in single cells.
  JOURNAL   Elife. 2022 Jun 9;11. pii: 75115. doi: 10.7554/eLife.75115.
  PUBMED    35678392
REFERENCE   2  (bases 1 to 12573)
  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: "Elife."; 
            date: "2022-06-9"; volume: "11. pii: 75115. doi"; pages: " 
            10.7554/eLife.75115"
COMMENT     Sequence Label: 
            pMH014_AAVS1_puro-pA-HS4-9xTetO-pEF-H2B-mCitrine-HS4-pRSV-H2B-mCherr
            y
FEATURES             Location/Qualifiers
     source          1..12573
                     /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"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    1065..1868
                     /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    1875..1900
                     /label=SA
                     /note="splice acceptor site"
     CDS             1924..1977
                     /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             1987..2586
                     /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(1987..2006)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2483..2503
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2618..2635)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2624..2848
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    4199..4217
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4235..4253
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4271..4289
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4307..4325
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4343..4361
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4379..4397
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4415..4433
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4451..4469
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4487..4505
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     intron          4750..5688
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5645..5665
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        5721..5739
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             5830..6207
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      6226..6235
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6232..6234,6235..6237,6238..6948)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6277..6298)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6538..6557)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6885..6906
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6999..7018)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7000..7018)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    7292..7413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        8712..8974
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     CDS             8988..9365
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      9378..9387
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             9384..10097
                     /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
                     EGRHSTGGMDELYKS"
     primer_bind     complement(9529..9547)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     9797..9816
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    10213..10294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        10337..10400
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    10479..11315
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     CDS             complement(join(11590..12381,12382..12450))
                     /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     12213..12232
                     /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 tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctca gaattaaccc
     1021 tcactaaagg gactagtcct gcaggtttaa acgaattcgc cctttgcttt ctctgaccag
     1081 cattctctcc cctgggcctg tgccgctttc tgtctgcagc ttgtggcctg ggtcacctct
     1141 acggctggcc cagatccttc cctgccgcct ccttcaggtt ccgtcttcct ccactccctc
     1201 ttccccttgc tctctgctgt gttgctgccc aaggatgctc tttccggagc acttccttct
     1261 cggcgctgca ccacgtgatg tcctctgagc ggatcctccc cgtgtctggg tcctctccgg
     1321 gcatctctcc tccctcaccc aaccccatgc cgtcttcact cgctgggttc ccttttcctt
     1381 ctccttctgg ggcctgtgcc atctctcgtt tcttaggatg gccttctccg acggatgtct
     1441 cccttgcgtc ccgcctcccc ttcttgtagg cctgcatcat caccgttttt ctggacaacc
     1501 ccaaagtacc ccgtctccct ggctttagcc acctctccat cctcttgctt tctttgcctg
     1561 gacaccccgt tctcctgtgg attcgggtca cctctcactc ctttcatttg ggcagctccc
     1621 ctacccccct tacctctcta gtctgtgcta gctcttccag ccccctgtca tggcatcttc
     1681 caggggtccg agagctcagc tagtcttctt cctccaaccc gggcccctat gtccacttca
     1741 ggacagcatg tttgctgcct ccagggatcc tgtgtccccg agctgggacc accttatatt
     1801 cccagggccg gttaatgtgg ctctggttct gggtactttt atctgtcccc tccaccccac
     1861 agtggggcaa gcttctgacc tcttctcttc ctcccacagg gcctcgagag atctggcagc
     1921 ggagagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctagg
     1981 ctcgagatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     2041 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     2101 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     2161 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     2221 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     2281 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     2341 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     2401 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     2461 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     2521 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     2581 gcctgatcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     2641 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2701 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2761 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2821 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctgggggcta
     2881 gcaaagcctt cgtttcgata agctctagag ggacagcccc ccccccaaag cccccaggga
     2941 tgtaattacg tccctccccc gctagggggc agcagcgagc cgcccggggc tccgctccgg
     3001 tccggcgctc cccccgcatc cccgagccgg cagcgtgcgg ggacagcccg ggcacgggga
     3061 aggtggcacg ggatcgcttt cctctgaacg cttctcgctg ctctttgagc ctgcagacac
     3121 ctggggggat acggggaaaa agctttaggc tgaaagagag atttagaatg acagaatcat
     3181 agaacggcct gggttgcaaa ggagcacagt gctcatccag atccaacccc ctgctatgtg
     3241 cagggtcatc aaccagcagc ccaggctgcc cagagccaca tccagcctgg ccttgaatgc
     3301 ctgcagggat ggggcatcca cagcctcctt gggcaacctg ttcagtgcgt caccaccctc
     3361 tgggggaaaa actgcctcct catatccaac ccaaacctcc cctgtctcag tgtaaagcca
     3421 ttcccccttg tcctatcaag ggggagtttg ctgtgacatt gttggtctgg ggtgacacat
     3481 gtttgccaat tcagtgcatc acggagaggc agatcttggg gataaggaag tgcaggacag
     3541 catggacgtg ggacatgcag gtgttgaggg ctctgggaca ctctccaagt cacagcgttc
     3601 agaacagcct taaggataag aagataggat agaaggacaa agagcaagtt aaaacccagc
     3661 atggagagga gcacaaaaag gccacagaca ctgctggtcc ctgtgtctga gcctgcatgt
     3721 ttgatggtgt ctggatgcaa gcagaagggg tggaagagct tgcctggaga gatacagctg
     3781 ggtcagtagg actgggacag gcagctggag aattgccatg tagatgttca tacaatcgtc
     3841 aaatcatgaa ggctggaaaa gccctccaag atccccaaga ccaaccccaa cccacccacc
     3901 gtgcccactg gccatgtccc tcagtgccac atccccacag ttcttcatca cctccaggga
     3961 cggtgacccc cccacctccg tgggcagctg tgccactgca gcaccgctct ttggagaagg
     4021 taaatcttgc taaatccagc ccgaccctcc cctggcacaa cgtaaggcca ttatctctca
     4081 tccaactcca ggacggagtc agtgaggatg gggctctagc ggggggatca aacggccggc
     4141 ctctagagtc gagatgacat tgattattga aacagctgca tatagcactg cagtttactc
     4201 cctatcagtg atagagagtc aaaaggccat gccctcccta tcagtgatag agagagcaga
     4261 ctcaaggtgg tccctatcag tgatagagag attgagcaag agggcctccc tatcagtgat
     4321 agagaatagc actgcagttt actccctatc agtgatagag agtcaaaagg ccatgccctc
     4381 cctatcagtg atagagagag cagactcaag gtggtcccta tcagtgatag agagattgag
     4441 caagagggcc tccctatcag tgatagagat tgatccagtc agggcctccc tatcagtgat
     4501 agagaagcta tgtgcaggta gggctgtaca gtgggaatgc atgagcgcac atcgcccaca
     4561 gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc
     4621 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
     4681 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
     4741 agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc
     4801 ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg tgattcttga tcccgagctt
     4861 cgggttggaa gtgggtggga gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt
     4921 gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt
     4981 cgcgcctgtc tcgctgcttt cgataagtct ctagccattt aaaatttttg atgacctgct
     5041 gcgacgcttt ttttctggca agatagtctt gtaaatgcgg gccaagatct gcacactggt
     5101 atttcggttt ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg
     5161 gcgaggcggg gcctgcgagc gcggccaccg agaatcggac gggggtagtc tcaagctggc
     5221 cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg
     5281 ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca
     5341 gggagctcaa aatggaggac gcggcgctcg ggagagcggg cgggtgagtc acccacacaa
     5401 aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg actccacgga gtaccgggcg
     5461 ccgtccaggc acctcgatta gttctcgagc ttttggagta cgtcgtcttt aggttggggg
     5521 gaggggtttt atgcgatgga gtttccccac actgagtggg tggagactga agttaggcca
     5581 gcttggcact tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg atcttggttc
     5641 attctcaagc ctcagacagt ggttcaaagt ttttttcttc catttcaggt gtcgtgagct
     5701 agcgattacg ccaagctcga aattaaccct cactaaaggg aacaaaagct ggagctccac
     5761 cgcggtggcg gccgctctag cccgggcgga tcccccgggc tgcaggaatt cgatatcaag
     5821 cttgccacca tgccagagcc agcgaagtct gctcccgccc cgaaaaaggg ctccaagaag
     5881 gcggtgacta aggcgcagaa gaaaggcggc aagaagcgca agcgcagccg caaggagagc
     5941 tattccatct atgtgtacaa ggttctgaag caggtccacc ctgacaccgg catttcgtcc
     6001 aaggccatgg gcatcatgaa ttcgtttgtg aacgacattt tcgagcgcat cgcaggtgag
     6061 gcttcccgcc tggcgcatta caacaagcgc tcgaccatca cctccaggga gatccagacg
     6121 gccgtgcgcc tgctgctgcc tggggagttg gccaagcacg ccgtgtccga gggtactaag
     6181 gccatcacca agtacaccag cgctaaggat ccccgggtac cggtcgccac catggtgagc
     6241 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     6301 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     6361 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     6421 accttcggct acggcctgat gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     6481 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     6541 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     6601 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     6661 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     6721 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     6781 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     6841 taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     6901 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta ggtcgacctc
     6961 gagggggggc ccggtaccag gtaagtgtac ccaattcgcc ctatagtgag tcgtattaca
     7021 attcactcga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg agatccaatt
     7081 tttaagtgta taatgtgtta aactactgat tctaattgtt tgtgtatttt agattcacag
     7141 tcccaaggct catttcaggc ccctcagtcc tcacagtctg ttcatgatca taatcagcca
     7201 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     7261 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     7321 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     7381 ttgtggtttg tccaaactca tcaatgtatc ttaacgtttc gataagctct agagggacag
     7441 ccccccccca aagcccccag ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg
     7501 agccgcccgg ggctccgctc cggtccggcg ctccccccgc atccccgagc cggcagcgtg
     7561 cggggacagc ccgggcacgg ggaaggtggc acgggatcgc tttcctctga acgcttctcg
     7621 ctgctctttg agcctgcaga cacctggggg gatacgggga aaaagcttta ggctgaaaga
     7681 gagatttaga atgacagaat catagaacgg cctgggttgc aaaggagcac agtgctcatc
     7741 cagatccaac cccctgctat gtgcagggtc atcaaccagc agcccaggct gcccagagcc
     7801 acatccagcc tggccttgaa tgcctgcagg gatggggcat ccacagcctc cttgggcaac
     7861 ctgttcagtg cgtcaccacc ctctggggga aaaactgcct cctcatatcc aacccaaacc
     7921 tcccctgtct cagtgtaaag ccattccccc ttgtcctatc aagggggagt ttgctgtgac
     7981 attgttggtc tggggtgaca catgtttgcc aattcagtgc atcacggaga ggcagatctt
     8041 ggggataagg aagtgcagga cagcatggac gtgggacatg caggtgttga gggctctggg
     8101 acactctcca agtcacagcg ttcagaacag ccttaaggat aagaagatag gatagaagga
     8161 caaagagcaa gttaaaaccc agcatggaga ggagcacaaa aaggccacag acactgctgg
     8221 tccctgtgtc tgagcctgca tgtttgatgg tgtctggatg caagcagaag gggtggaaga
     8281 gcttgcctgg agagatacag ctgggtcagt aggactggga caggcagctg gagaattgcc
     8341 atgtagatgt tcatacaatc gtcaaatcat gaaggctgga aaagccctcc aagatcccca
     8401 agaccaaccc caacccaccc accgtgccca ctggccatgt ccctcagtgc cacatcccca
     8461 cagttcttca tcacctccag ggacggtgac ccccccacct ccgtgggcag ctgtgccact
     8521 gcagcaccgc tctttggaga aggtaaatct tgctaaatcc agcccgaccc tcccctggca
     8581 caacgtaagg ccattatctc tcatccaact ccaggacgga gtcagtgagg atggggctct
     8641 agcgggggga tcaaacggcc ggcctctaga gtcgagatga cattgattat tgacctaggg
     8701 acgtccatat gaatgtagtc ttatgcaata cacttgtagt cttgcaacat ggtaacgatg
     8761 agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt ggtggaagta
     8821 aggtggtacg atcgtgcctt attaggaagg caacagacag gtctgacatg gattggacga
     8881 accactgaat tgcgcattgc agagataatt gtatttaagt gcctagctcg atacaataaa
     8941 cgccatttga ccattcacca cattggtgtg caccctgcag atctagaatg ccagagccag
     9001 cgaagtctgc tcccgccccg aaaaagggct ccaagaaggc ggtgactaag gcgcagaaga
     9061 aaggcggcaa gaagcgcaag cgcagccgca aggagagcta ttccatctat gtgtacaagg
     9121 ttctgaagca ggtccaccct gacaccggca tttcgtccaa ggccatgggc atcatgaatt
     9181 cgtttgtgaa cgacattttc gagcgcatcg caggtgaggc ttcccgcctg gcgcattaca
     9241 acaagcgctc gaccatcacc tccagggaga tccagacggc cgtgcgcctg ctgctgcctg
     9301 gggagttggc caagcacgcc gtgtccgagg gtactaaggc catcaccaag tacaccagcg
     9361 ctaaggatcc accggtcgcc accatggtga gcaagggcga ggaggataac atggccatca
     9421 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
     9481 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     9541 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
     9601 gctccaaggc ctacgtgaag caccccgccg acattcccga ctacttgaag ctgtccttcc
     9661 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     9721 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     9781 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     9841 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     9901 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     9961 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
    10021 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
    10081 agctgtacaa gtcttaagcg actctagatc ataatcagcc ataccacatt tgtagaggtt
    10141 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
    10201 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
    10261 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
    10321 atcaatgtat ctcatatgtg tatccgctca tgagacaata accctgataa atgcttcaat
    10381 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctaaa
    10441 ttgtaagcgt ctcggcctct agagacattg attattgaac tagggacagg attggtgaca
    10501 gaaaagcccc atccttaggc ctcctccttc ctagtctcct gatattgggt ctaaccccca
    10561 cctcctgtta ggcagattcc ttatctggtg acacaccccc atttcctgga gccatctctc
    10621 tccttgccag aacctctaag gtttgcttac gatggagcca gagaggatcc tgggagggag
    10681 agcttggcag ggggtgggag ggaagggggg gatgcgtgac ctgcccggtt ctcagtggcc
    10741 accctgcgct accctctccc agaacctgag ctgctctgac gcggctgtct ggtgcgtttc
    10801 actgatcctg gtgctgcagc ttccttacac ttcccaagag gagaagcagt ttggaaaaac
    10861 aaaatcagaa taagttggtc ctgagttcta actttggctc ttcacctttc tagtccccaa
    10921 tttatattgt tcctccgtgc gtcagtttta cctgtgagat aaggccagta gccagccccg
    10981 tcctggcagg gctgtggtga ggaggggggt gtccgtgtgg aaaactccct ttgtgagaat
    11041 ggtgcgtcct aggtgttcac caggtcgtgg ccgcctctac tccctttctc tttctccatc
    11101 cttctttcct taaagagtcc ccagtgctat ctgggacata ttcctccgcc cagagcaggg
    11161 tcccgcttcc ctaaggccct gctctgggct tctgggtttg agtccttggc aagcccagga
    11221 gaggcgctca ggcttccctg tcccccttcc tcgtccacca tctcatgccc ctggctctcc
    11281 tgccccttcc ctacaggggt tcctggctct gctcttcgcc ttcttgacga gttcttctga
    11341 attattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
    11401 tcacaccgca tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
    11461 tttctaaata cattcaaata tgtatccgct catgagatta tcaaaaagga tcttcaccta
    11521 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
    11581 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
    11641 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
    11701 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
    11761 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
    11821 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
    11881 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
    11941 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
    12001 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
    12061 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
    12121 atgcttttct gtgactggtg aatactcaac caagtcattc tgagaatagt gtatgcggcg
    12181 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
    12241 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
    12301 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
    12361 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
    12421 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
    12481 ttatcagggt tattgtctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
    12541 gtcagacccc gtagaaaaga tcaaaggatc ttc
//
