LOCUS       sequence-355448-       14964 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual fluorescent reporter construct with 5kb lambda spacer, 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 14964)
  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 14964)
  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: 
            pSL012_AAVS1-puro-9xTetO-pEF-H2B-mCitrine-5kb,lambda-pRSV-H2B-mCherr
            y
FEATURES             Location/Qualifiers
     source          1..14964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    7..32
                     /label=SA
                     /note="splice acceptor site"
     CDS             56..109
                     /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             119..718
                     /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(119..138)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     615..635
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(750..767)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    756..980
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    1053..1071
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1089..1107
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1125..1143
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1161..1179
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1197..1215
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1233..1251
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1269..1287
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1305..1323
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1341..1359
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     intron          1604..2542
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2499..2519
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        2575..2593
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             2684..3061
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      3080..3089
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3086..3088,3089..3091,3092..3802)
                     /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(3131..3152)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3392..3411)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3739..3760
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3853..3872)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3854..3872)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4146..4267
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    4873..5104
                     /gene="attP"
                     /label=lambda attP
                     /bound_moiety="lambda integrase"
                     /note="integrase from phage lambda"
     CDS             complement(5099..6169)
                     /codon_start=1
                     /gene="int"
                     /product="integrase from phage lambda"
                     /label=lambda integrase
                     /translation="MGRRRSHERRDLPPNLYIRNNGYYCYRDPRTGKEFGLGRDRRIAI
                     TEAIQANIELFSGHKHKPLTARINSDNSVTLHSWLDRYEKILASRGIKQKTLINYMSKI
                     KAIRRGLPDAPLEDITTKEIAAMLNGYIDEGKAASAKLIRSTLSDAFREAIAEGHITTN
                     HVAATRAAKSEVRRSRLTADEYLKIYQAAESSPCWLRLAMELAVVTGQRVGDLCEMKWS
                     DIVDGYLYVEQSKTGVKIAIPTALHIDALGISMKETLDKCKEILGGETIIASTRREPLS
                     SGTVSRYFMRARKASGLSFEGDPPTFHELRSLSARLYEKQISDKFAQHLLGHKSDTMAS
                     QYRDDRGREWDKTEIK"
     terminator      complement(8390..8634)
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     promoter        9325..9587
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     CDS             9601..9978
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      9991..10000
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             9997..10710
                     /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(10142..10160)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     10410..10429
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    10826..10907
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    11002..11838
                     /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(12113..12904,12905..12973))
                     /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     12736..12755
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      13097..13685
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13586..13605
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13839..13856
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13973..13994
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(14009..14014,14015..14032,14033..14039)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14047..14063
                     /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     14052..14074
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     14071..14087
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     14071..14087
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        14108..14126
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    14161..14964
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
ORIGIN
        1 aagcttctga cctcttctct tcctcccaca gggcctcgag agatctggca gcggagaggg
       61 cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggcccta ggctcgagat
      121 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
      181 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
      241 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
      301 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
      361 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
      421 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
      481 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
      541 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
      601 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
      661 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgatc
      721 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
      781 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
      841 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
      901 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
      961 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctgggggc tagcaaagcc
     1021 taggaacagc tgcatatagc actgcagttt actccctatc agtgatagag agtcaaaagg
     1081 ccatgccctc cctatcagtg atagagagag cagactcaag gtggtcccta tcagtgatag
     1141 agagattgag caagagggcc tccctatcag tgatagagaa tagcactgca gtttactccc
     1201 tatcagtgat agagagtcaa aaggccatgc cctccctatc agtgatagag agagcagact
     1261 caaggtggtc cctatcagtg atagagagat tgagcaagag ggcctcccta tcagtgatag
     1321 agattgatcc agtcagggcc tccctatcag tgatagagaa gctatgtgca ggtagggctg
     1381 tacagtggga atgcatgagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
     1441 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
     1501 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
     1561 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg
     1621 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac
     1681 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc
     1741 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg
     1801 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa
     1861 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag
     1921 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg
     1981 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc
     2041 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc
     2101 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg
     2161 agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg
     2221 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc
     2281 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc
     2341 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc
     2401 ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt
     2461 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca
     2521 aagttttttt cttccatttc aggtgtcgtg agctagcgat tacgccaagc tcgaaattaa
     2581 ccctcactaa agggaacaaa agctggagct ccaccgcggt ggcggccgct ctagcccggg
     2641 cggatccccc gggctgcagg aattcgatat caagcttgcc accatgccag agccagcgaa
     2701 gtctgctccc gccccgaaaa agggctccaa gaaggcggtg actaaggcgc agaagaaagg
     2761 cggcaagaag cgcaagcgca gccgcaagga gagctattcc atctatgtgt acaaggttct
     2821 gaagcaggtc caccctgaca ccggcatttc gtccaaggcc atgggcatca tgaattcgtt
     2881 tgtgaacgac attttcgagc gcatcgcagg tgaggcttcc cgcctggcgc attacaacaa
     2941 gcgctcgacc atcacctcca gggagatcca gacggccgtg cgcctgctgc tgcctgggga
     3001 gttggccaag cacgccgtgt ccgagggtac taaggccatc accaagtaca ccagcgctaa
     3061 ggatccccgg gtaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
     3121 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     3181 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     3241 caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgatgtgctt
     3301 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     3361 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     3421 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     3481 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     3541 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     3601 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     3661 ccccgtgctg ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc
     3721 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     3781 cggcatggac gagctgtaca agtaggtcga cctcgagggg gggcccggta ccaggtaagt
     3841 gtacccaatt cgccctatag tgagtcgtat tacaattcac tcgatcgccc ttcccaacag
     3901 ttgcgcagcc tgaatggcga atggagatcc aatttttaag tgtataatgt gttaaactac
     3961 tgattctaat tgtttgtgta ttttagattc acagtcccaa ggctcatttc aggcccctca
     4021 gtcctcacag tctgttcatg atcataatca gccataccac atttgtagag gttttacttg
     4081 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     4141 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4201 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4261 tatcttaacg cgtaaattgt aagcgtctca gcaatttatt tggcggcaac acaggatctc
     4321 tcttttaagt tactctctat tacatacgtt ttccatctaa aaattagtag tattgaactt
     4381 aacggggcat cgtattgtag ttttccatat ttagctttct gcttcctttt ggataaccca
     4441 ctgttattca tgttgcatgg tgcactgttt ataccaacga tatagtctat taatgcatat
     4501 atagtatcgc cgaacgatta gctcttcagg cttctgaaga agcgtttcaa gtactaataa
     4561 gccgatagat agccacggac ttcgtagcca tttttcataa gtgttaactt ccgctcatcg
     4621 ctcataacag acattcacta cagttatggc ggaaaggtat gcatgctggg tgtggggaag
     4681 tcgtgaaaga aaagaagtca gctgcgtcgt ttgacatcac tgctatcttc ttactggtta
     4741 tgcaggtcgt agtgggtggc acacaaagct ttgcactgga ttgcgaggct ttgtgcttct
     4801 ctggagtgcg acaggtttga tgacaaaaaa ttagcgcaag aagacaaaaa tcaccttgcg
     4861 ctaatgctct gttacaggtc actaatacca tctaagtagt tgattcatag tgactgcata
     4921 tgttgtgttt tacagtatta tgtagtctgt tttttatgca aaatctaatt taatatattg
     4981 atatttatat cattttacgt ttctcgttca gcttttttat actaagttgg cattataaaa
     5041 aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt
     5101 atttgatttc agttttgtcc cactccctgc ctctgtcatc acgatactgt gatgccatgg
     5161 tgtccgactt atgcccgaga agatgttgag caaacttatc gcttatctgc ttctcataga
     5221 gtcttgcaga caaactgcgc aactcgtgaa aggtaggcgg atccccttcg aaggaaagac
     5281 ctgatgcttt tcgtgcgcgc ataaaatacc ttgatactgt gccggatgaa agcggttcgc
     5341 gacgagtaga tgcaattatg gtttctccgc caagaatctc tttgcattta tcaagtgttt
     5401 ccttcattga tattccgaga gcatcaatat gcaatgctgt tgggatggca atttttacgc
     5461 ctgttttgct ttgctcgaca taaagatatc catctacgat atcagaccac ttcatttcgc
     5521 ataaatcacc aactcgttgc ccggtaacaa cagccagttc cattgcaagt ctgagccaac
     5581 atggtgatga ttctgctgct tgataaattt tcaggtattc gtcagccgta agtcttgatc
     5641 tccttacctc tgattttgct gcgcgagtgg cagcgacatg gtttgttgtt atatggcctt
     5701 cagctattgc ctctcggaat gcatcgctca gtgttgatct gattaacttg gctgacgccg
     5761 ccttgccctc gtctatgtat ccattgagca ttgccgcaat ttcttttgtg gtgatgtctt
     5821 caagtggagc atcaggcaga cccctcctta ttgctttaat tttgctcatg taatttatga
     5881 gtgtcttctg cttgattcct ctgctggcca ggattttttc gtagcgatca agccatgaat
     5941 gtaacgtaac ggaattatca ctgttgattc tcgctgtcag aggcttgtgt ttgtgtcctg
     6001 aaaataactc aatgttggcc tgtatagctt cagtgattgc gattcgcctg tctctgccta
     6061 atccaaactc tttacccgtc cttgggtccc tgtagcagta atatccattg tttcttatat
     6121 aaaggttagg gggtaaatcc cggcgctcat gacttcgcct tcttcccatt tctgatcctc
     6181 ttcaaaaggc cacctgttac tggtcgattt aagtcaacct ttaccgctga ttcgtggaac
     6241 agatactctc ttccatcctt aaccggaggt gggaatatcc tgcattcccg aacccatcga
     6301 cgaactgttt caaggcttct tggacgtcgc tggcgtgcgt tccactcctg aagtgtcaag
     6361 tacatcgcaa agtctccgca attacacgca agaaaaaacc gccatcaggc ggcttggtgt
     6421 tctttcagtt cttcaattcg aatattggtt acgtctgcat gtgctatctg cgcccatatc
     6481 atccagtggt cgtagcagtc gttgatgttc tccgcttcga taactctgtt gaatggctct
     6541 ccattccatt ctcctgtgac tcggaagtgc atttatcatc tccataaaac aaaacccgcc
     6601 gtagcgagtt cagataaaat aaatccccgc gagagcgagg attgttatgt aatattgggt
     6661 ttaatcatct atatgttttg tacagagagg gcaagtatcg tttccaccgt actcgtgata
     6721 ataattttgc acggtatcag tcatttctcg cacattgcag aatggggatt tgtcttcatt
     6781 agacttataa accttcatgg aatatttgta tgccgactct atatctatac cttcatctac
     6841 ataaacacct tcgtgatgtc tgcatggaga caagacaccg gatctgcaca acattgataa
     6901 cgcccaatct ttttgctcag actctaactc attgatactc atttataaac tccttgcaat
     6961 gtatgtcgtt tcagctaaac ggtatcagca atgtttatgt aaagaaacag taagataata
     7021 ctcaacccga tgtttgagta cggtcatcat ctgacactac agactctggc atcgctgtga
     7081 agacgacgcg aaattcagca ttttcacaag cgttatcttt tacaaaaccg atctcactct
     7141 cctttgatgc gaatgccagc gtcagacatc atatgcagat actcacctgc atcctgaacc
     7201 cattgacctc caaccccgta atagcgatgc gtaatgatgt cgatagttac taacgggtct
     7261 tgttcgatta actgccgcag aaactcttcc aggtcaccag tgcagcgctt gataacagga
     7321 gtcttcccag gatggcgaac aacaagaaac tggtttccgt cttcacggac ttcgttgctt
     7381 tccagtttag caatacgctt actcccatcc gagataacac cttcgtaata ctcacgctgc
     7441 tcgttgagtt ttgattttgc tgtttcaagc tcaacacgca gtttccctac tgttagcgca
     7501 atatcctcgt tctcctggtc gcggcgtttg atgtattgct ggtttctttc ccgttcatcc
     7561 agcagttcca gcacaatcga tggtgttacc aattcatgga aaaggtctgc gtcaaatccc
     7621 cagtcgtcat gcattgcctg ctctgccgct tcacgcagtg cctgagagtt aatttcgctc
     7681 acttcgaacc tctctgttta ctgataagtt ccagatcctc ctggcaactt gcacaagtcc
     7741 gacaaccctg aacgaccagg cgtcttcgtt catctatcgg atcgccacac tcacaacaat
     7801 gagtggcaga tatagcctgg tggttcaggc ggcgcatttt tattgctgtg ttgcgctgta
     7861 attcttctat ttctgatgct gaatcaatga tgtctgccat ctttcattaa tccctgaact
     7921 gttggttaat acgcttgagg gtgaatgcga ataataaaaa aggagcctgt agctccctga
     7981 tgattttgct tttcatgttc atcgttcctt aaagacgccg tttaacatgc cgattgccag
     8041 gcttaaatga gtcggtgtga atcccatcag cgttaccgtt tcgcggtgct tcttcagtac
     8101 gctacggcaa atgtcatcga cgtttttatc cggaaactgc tgtctggctt tttttgattt
     8161 cagaattagc ctgacgggca atgctgcgaa gggcgttttc ctgctgaggt gtcattgaac
     8221 aagtcccatg tcggcaagca taagcacaca gaatatgaag cccgctgcca gaaaaatgca
     8281 ttccgtggtt gtcatacctg gtttctctca tctgcttctg ctttcgccac catcatttcc
     8341 agcttttgtg aaagggatgc ggctaacgta tgaaattctt cgtctgtttc tactggtatt
     8401 ggcacaaacc tgattccaat ttgagcaagg ctatgtgcca tctcgatact cgttcttaac
     8461 tcaacagaag atgctttgtg catacagccc ctcgtttatt atttatctcc tcagccagcc
     8521 gctgtgcttt cagtggattt cggataacag aaaggccggg aaatacccag cctcgctttg
     8581 taacggagta gacgaaagtg attgcgccta cccggatatt atcgtgagga tgcgtcatcg
     8641 ccattgctcc ccaaatacaa aaccaatttc agccagtgcc tcgtccattt tttcgatgaa
     8701 ctccggcacg atctcgtcaa aactcgccat gtacttttca tcccgctcaa tcacgacata
     8761 atgcaggcct tcacgcttca tacgcgggtc atagttggca aagtaccagg cattttttcg
     8821 cgtcacccac atgctgtact gcacctgggc catgtaagct gactttatgg cctcgaaacc
     8881 accgagccgg aacttcatga aatcccggga ggtaaacggg catttcagtt caaggccgtt
     8941 gccgtcactg cataaaccat cgggagagca ggcggtacgc atactttcgt cgcgatagat
     9001 gatcggggat tcagtaacat tcacgccgga agtgaattca aacagggttc tggcgtcgtt
     9061 ctcgtactgt tttccccagg ccagtgcttt agcgttaact tccggagcca caccggtgca
     9121 aacctcagca agcagggtgt ggaagtagga cattttcatg tcaggccact tctttccgga
     9181 gcggggtttt gctatcacgt tgtgaacttc tgaagcggtg atgacgccga gccgtaattt
     9241 gtgccacgca tcatccccct gttcgacagc tctcacatcg atcccggtac gctgcaggat
     9301 aatgtcccta gggacgtcca tatgaatgta gtcttatgca atacacttgt agtcttgcaa
     9361 catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc gtgcatgccg
     9421 attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga caggtctgac
     9481 atggattgga cgaaccactg aattgcgcat tgcagagata attgtattta agtgcctagc
     9541 tcgatacaat aaacgccatt tgaccattca ccacattggt gtgcaccctg cagatctaga
     9601 atgccagagc cagcgaagtc tgctcccgcc ccgaaaaagg gctccaagaa ggcggtgact
     9661 aaggcgcaga agaaaggcgg caagaagcgc aagcgcagcc gcaaggagag ctattccatc
     9721 tatgtgtaca aggttctgaa gcaggtccac cctgacaccg gcatttcgtc caaggccatg
     9781 ggcatcatga attcgtttgt gaacgacatt ttcgagcgca tcgcaggtga ggcttcccgc
     9841 ctggcgcatt acaacaagcg ctcgaccatc acctccaggg agatccagac ggccgtgcgc
     9901 ctgctgctgc ctggggagtt ggccaagcac gccgtgtccg agggtactaa ggccatcacc
     9961 aagtacacca gcgctaagga tccaccggtc gccaccatgg tgagcaaggg cgaggaggat
    10021 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
    10081 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
    10141 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
    10201 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacattcc cgactacttg
    10261 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
    10321 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
    10381 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
    10441 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
    10501 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
    10561 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
    10621 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
    10681 ggcggcatgg acgagctgta caagtcttaa gcgactctag atcataatca gccataccac
    10741 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
    10801 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
    10861 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
    10921 tttgtccaaa ctcatcaatg tatctcatat gtaaattgta agcgtctcgg cctctagagt
    10981 cgagatgaca ttgattattg aactagggac aggattggtg acagaaaagc cccatcctta
    11041 ggcctcctcc ttcctagtct cctgatattg ggtctaaccc ccacctcctg ttaggcagat
    11101 tccttatctg gtgacacacc cccatttcct ggagccatct ctctccttgc cagaacctct
    11161 aaggtttgct tacgatggag ccagagagga tcctgggagg gagagcttgg cagggggtgg
    11221 gagggaaggg ggggatgcgt gacctgcccg gttctcagtg gccaccctgc gctaccctct
    11281 cccagaacct gagctgctct gacgcggctg tctggtgcgt ttcactgatc ctggtgctgc
    11341 agcttcctta cacttcccaa gaggagaagc agtttggaaa aacaaaatca gaataagttg
    11401 gtcctgagtt ctaactttgg ctcttcacct ttctagtccc caatttatat tgttcctccg
    11461 tgcgtcagtt ttacctgtga gataaggcca gtagccagcc ccgtcctggc agggctgtgg
    11521 tgaggagggg ggtgtccgtg tggaaaactc cctttgtgag aatggtgcgt cctaggtgtt
    11581 caccaggtcg tggccgcctc tactcccttt ctctttctcc atccttcttt ccttaaagag
    11641 tccccagtgc tatctgggac atattcctcc gcccagagca gggtcccgct tccctaaggc
    11701 cctgctctgg gcttctgggt ttgagtcctt ggcaagccca ggagaggcgc tcaggcttcc
    11761 ctgtccccct tcctcgtcca ccatctcatg cccctggctc tcctgcccct tccctacagg
    11821 ggttcctggc tctgctcttc gccttcttga cgagttcttc tgaattatta acgcttacaa
    11881 tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatcaggtg
    11941 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
    12001 atatgtatcc gctcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
    12061 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
    12121 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
    12181 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
    12241 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
    12301 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
    12361 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
    12421 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
    12481 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
    12541 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
    12601 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
    12661 gtgaatactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
    12721 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
    12781 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
    12841 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
    12901 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
    12961 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
    13021 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
    13081 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
    13141 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
    13201 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
    13261 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
    13321 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
    13381 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
    13441 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
    13501 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
    13561 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
    13621 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
    13681 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
    13741 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
    13801 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
    13861 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
    13921 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
    13981 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
    14041 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
    14101 agctcagaat taaccctcac taaagggact agtcctgcag gtttaaacga attcgccctt
    14161 tgctttctct gaccagcatt ctctcccctg ggcctgtgcc gctttctgtc tgcagcttgt
    14221 ggcctgggtc acctctacgg ctggcccaga tccttccctg ccgcctcctt caggttccgt
    14281 cttcctccac tccctcttcc ccttgctctc tgctgtgttg ctgcccaagg atgctctttc
    14341 cggagcactt ccttctcggc gctgcaccac gtgatgtcct ctgagcggat cctccccgtg
    14401 tctgggtcct ctccgggcat ctctcctccc tcacccaacc ccatgccgtc ttcactcgct
    14461 gggttccctt ttccttctcc ttctggggcc tgtgccatct ctcgtttctt aggatggcct
    14521 tctccgacgg atgtctccct tgcgtcccgc ctccccttct tgtaggcctg catcatcacc
    14581 gtttttctgg acaaccccaa agtaccccgt ctccctggct ttagccacct ctccatcctc
    14641 ttgctttctt tgcctggaca ccccgttctc ctgtggattc gggtcacctc tcactccttt
    14701 catttgggca gctcccctac cccccttacc tctctagtct gtgctagctc ttccagcccc
    14761 ctgtcatggc atcttccagg ggtccgagag ctcagctagt cttcttcctc caacccgggc
    14821 ccctatgtcc acttcaggac agcatgtttg ctgcctccag ggatcctgtg tccccgagct
    14881 gggaccacct tatattccca gggccggtta atgtggctct ggttctgggt acttttatct
    14941 gtcccctcca ccccacagtg gggc
//
