LOCUS       sequence-355551-       10305 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Dual fluorescent reporter construct with single core 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 10305)
  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 10305)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: 
            pMH012_AAVS1_puro-pA-9xTetO-pEF-H2B-mCitrine-core-pRSV-H2B-mCherry
FEATURES             Location/Qualifiers
     source          1..10305
                     /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    2921..2939
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2957..2975
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2993..3011
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3029..3047
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3065..3083
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3101..3119
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3137..3155
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3173..3191
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3209..3227
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     intron          3472..4410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4367..4387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        4443..4461
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             4552..4929
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      4948..4957
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4954..4956,4957..4959,4960..5670)
                     /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(4999..5020)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5260..5279)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5607..5628
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5721..5740)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5722..5740)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    6014..6135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        6436..6698
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     CDS             6712..7089
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      7102..7111
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7108..7821
                     /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(7253..7271)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7521..7540
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    7937..8018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        8061..8124
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    8211..9047
                     /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(9322..10113,10114..10182))
                     /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     9945..9964
                     /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 cgaacagctg catatagcac tgcagtttac tccctatcag tgatagagag
     2941 tcaaaaggcc atgccctccc tatcagtgat agagagagca gactcaaggt ggtccctatc
     3001 agtgatagag agattgagca agagggcctc cctatcagtg atagagaata gcactgcagt
     3061 ttactcccta tcagtgatag agagtcaaaa ggccatgccc tccctatcag tgatagagag
     3121 agcagactca aggtggtccc tatcagtgat agagagattg agcaagaggg cctccctatc
     3181 agtgatagag attgatccag tcagggcctc cctatcagtg atagagaagc tatgtgcagg
     3241 tagggctgta cagtgggaat gcatgagcgc acatcgccca cagtccccga gaagttgggg
     3301 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
     3361 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
     3421 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
     3481 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
     3541 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
     3601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
     3661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
     3721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
     3781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
     3841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
     3901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
     3961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     4021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     4081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     4141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     4201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     4261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     4321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     4381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag ctagcgatta cgccaagctc
     4441 gaaattaacc ctcactaaag ggaacaaaag ctggagctcc accgcggtgg cggccgctct
     4501 agcccgggcg gatcccccgg gctgcaggaa ttcgatatca agcttgccac catgccagag
     4561 ccagcgaagt ctgctcccgc cccgaaaaag ggctccaaga aggcggtgac taaggcgcag
     4621 aagaaaggcg gcaagaagcg caagcgcagc cgcaaggaga gctattccat ctatgtgtac
     4681 aaggttctga agcaggtcca ccctgacacc ggcatttcgt ccaaggccat gggcatcatg
     4741 aattcgtttg tgaacgacat tttcgagcgc atcgcaggtg aggcttcccg cctggcgcat
     4801 tacaacaagc gctcgaccat cacctccagg gagatccaga cggccgtgcg cctgctgctg
     4861 cctggggagt tggccaagca cgccgtgtcc gagggtacta aggccatcac caagtacacc
     4921 agcgctaagg atccccgggt accggtcgcc accatggtga gcaagggcga ggagctgttc
     4981 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     5041 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     5101 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg
     5161 atgtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     5221 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     5281 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     5341 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     5401 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     5461 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     5521 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc
     5581 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     5641 atcactctcg gcatggacga gctgtacaag taggtcgacc tcgagggggg gcccggtacc
     5701 aggtaagtgt acccaattcg ccctatagtg agtcgtatta caattcactc gatcgccctt
     5761 cccaacagtt gcgcagcctg aatggcgaat ggagatccaa tttttaagtg tataatgtgt
     5821 taaactactg attctaattg tttgtgtatt ttagattcac agtcccaagg ctcatttcag
     5881 gcccctcagt cctcacagtc tgttcatgat cataatcagc cataccacat ttgtagaggt
     5941 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     6001 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6061 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6121 catcaatgta tcttaatctc aaagtaacaa aacttttatg agggacagcc cccccccaaa
     6181 gcccccaggg atgtaattac gtccctcccc cgctaggggg cagcagcgag ccgcccgggg
     6241 ctccgctccg gtccggcgct ccccccgcat ccccgagccg gcagcgtgcg gggacagccc
     6301 gggcacgggg aaggtggcac gggatcgctt tcctctgaac gcttctcgct gctctttgag
     6361 cctgcagaca cctgggggga tacggggaaa aggcctccac ggccactagt attatgccct
     6421 agggacgtcc atatgaatgt agtcttatgc aatacacttg tagtcttgca acatggtaac
     6481 gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga
     6541 agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acaggtctga catggattgg
     6601 acgaaccact gaattgcgca ttgcagagat aattgtattt aagtgcctag ctcgatacaa
     6661 taaacgccat ttgaccattc accacattgg tgtgcaccct gcagatctag aatgccagag
     6721 ccagcgaagt ctgctcccgc cccgaaaaag ggctccaaga aggcggtgac taaggcgcag
     6781 aagaaaggcg gcaagaagcg caagcgcagc cgcaaggaga gctattccat ctatgtgtac
     6841 aaggttctga agcaggtcca ccctgacacc ggcatttcgt ccaaggccat gggcatcatg
     6901 aattcgtttg tgaacgacat tttcgagcgc atcgcaggtg aggcttcccg cctggcgcat
     6961 tacaacaagc gctcgaccat cacctccagg gagatccaga cggccgtgcg cctgctgctg
     7021 cctggggagt tggccaagca cgccgtgtcc gagggtacta aggccatcac caagtacacc
     7081 agcgctaagg atccaccggt cgccaccatg gtgagcaagg gcgaggagga taacatggcc
     7141 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     7201 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     7261 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     7321 tacggctcca aggcctacgt gaagcacccc gccgacattc ccgactactt gaagctgtcc
     7381 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     7441 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     7501 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     7561 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     7621 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     7681 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     7741 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     7801 gacgagctgt acaagtctta agcgactcta gatcataatc agccatacca catttgtaga
     7861 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     7921 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     7981 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     8041 actcatcaat gtatctcata tgtgtatccg ctcatgagac aataaccctg ataaatgctt
     8101 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     8161 taaattgtaa gcgtctcggc ctctagagtc gagatgacat tgattattga actagggaca
     8221 ggattggtga cagaaaagcc ccatccttag gcctcctcct tcctagtctc ctgatattgg
     8281 gtctaacccc cacctcctgt taggcagatt ccttatctgg tgacacaccc ccatttcctg
     8341 gagccatctc tctccttgcc agaacctcta aggtttgctt acgatggagc cagagaggat
     8401 cctgggaggg agagcttggc agggggtggg agggaagggg gggatgcgtg acctgcccgg
     8461 ttctcagtgg ccaccctgcg ctaccctctc ccagaacctg agctgctctg acgcggctgt
     8521 ctggtgcgtt tcactgatcc tggtgctgca gcttccttac acttcccaag aggagaagca
     8581 gtttggaaaa acaaaatcag aataagttgg tcctgagttc taactttggc tcttcacctt
     8641 tctagtcccc aatttatatt gttcctccgt gcgtcagttt tacctgtgag ataaggccag
     8701 tagccagccc cgtcctggca gggctgtggt gaggaggggg gtgtccgtgt ggaaaactcc
     8761 ctttgtgaga atggtgcgtc ctaggtgttc accaggtcgt ggccgcctct actccctttc
     8821 tctttctcca tccttctttc cttaaagagt ccccagtgct atctgggaca tattcctccg
     8881 cccagagcag ggtcccgctt ccctaaggcc ctgctctggg cttctgggtt tgagtccttg
     8941 gcaagcccag gagaggcgct caggcttccc tgtccccctt cctcgtccac catctcatgc
     9001 ccctggctct cctgcccctt ccctacaggg gttcctggct ctgctcttcg ccttcttgac
     9061 gagttcttct gaattattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat
     9121 ctgtgcggta tttcacaccg catcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     9181 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagat tatcaaaaag
     9241 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     9301 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     9361 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     9421 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     9481 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     9541 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     9601 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     9661 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     9721 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     9781 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     9841 gccatccgta agatgctttt ctgtgactgg tgaatactca accaagtcat tctgagaata
     9901 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     9961 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
    10021 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
    10081 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
    10141 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
    10201 ttattgaagc atttatcagg gttattgtct catgaccaaa atcccttaac gtgagttttc
    10261 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
