LOCUS       sequence-169485-        9629 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Multi-gene expression system for purification of human box C/D RNPs 
            with U14-tRNA minigene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9629)
  AUTHORS   Peng Y, Yu G, Tian S, Li H
  TITLE     Co-expression and co-purification of archaeal and eukaryal box C/D 
            RNPs.
  JOURNAL   PLoS One. 2014 Jul 31;9(7):e103096. doi: 
            10.1371/journal.pone.0103096. eCollection 2014.
  PUBMED    25078083
REFERENCE   2  (bases 1 to 9629)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2014-07-31"; volume: "9(7):e103096. doi"; pages: " 
            10.1371/journal.pone.0103096. eCollection 2014"
COMMENT     Sequence Label: pQhsCDtRNA+
FEATURES             Location/Qualifiers
     source          1..9629
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             887..907
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     protein_bind    complement(924..948)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      970..1064
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        join(1093..1107,1108..1113,1114..1130,1131..1137)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    1113..1129
                     /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)."
     protein_bind    1145..1161
                     /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)."
     RBS             1180..1191
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    complement(2423..2447)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      2469..2563
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        join(2592..2606,2607..2612,2613..2629,2630..2636)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    2612..2628
                     /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)."
     protein_bind    2644..2660
                     /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)."
     RBS             2679..2690
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2706..2730
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3118..3142)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      3164..3258
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3352..4011
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3418..3437)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4076..4162
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4255..5337)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5299..5318
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5338..5415)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(5499..5521)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5607..5747
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5662..5681
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5762..5779)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5933..6521)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6013..6032)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6692..7483,7484..7552))
                     /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     7315..7334
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7553..7657)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7725..7743
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      8051..8145
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        join(8174..8188,8189..8194,8195..8211,8212..8218)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    8194..8210
                     /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)."
     protein_bind    8226..8242
                     /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)."
     RBS             8261..8272
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(8816..8827)
                     /codon_start=1
                     /gene="cspA"
                     /product="translation enhancing element for E. coli (Qing 
                     et al., 2004)"
                     /label=TEE
                     /translation="NHKV"
     protein_bind    complement(9003..9027)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      9049..9143
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        join(9172..9186,9187..9192,9193..9209,9210..9216)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    9192..9208
                     /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)."
     protein_bind    9224..9240
                     /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)."
     RBS             9259..9270
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 agtcatagct gagatcctgc gaggagttcg tctgcacttc cacaatctgg tgaagggtct
       61 gaccgatctg tcagcttgta aagcacagct ggggctggga cacagctatt cccgtgccaa
      121 agttaagttt aatgtgaacc gggtggacaa tatgatcatc cagtccatta gcctcctgga
      181 ccagctggat aaggacatca ataccttctc tatgcgtgtc agggagtggt acgggtatca
      241 ctttccggag ctggtgaaga tcatcaacga caatgccaca tactgccgtc ttgcccagtt
      301 tattggaaac cgaagggaac tgaatgagga caagctggag aagctggagg agctgacaat
      361 ggatggggcc aaggctaagg ctattctgga tgcctcacgg tcctccatgg gcatggacat
      421 atctgccatt gacttgataa acatcgagag cttctccagt cgtgtggtgt ctttatctga
      481 ataccgccag agcctacaca cttacctgcg ctccaagatg agccaagtag cccccagcct
      541 gtcagcccta attggggaag cggtaggtgc acgtctcatc gcacatgctg gcagcctcac
      601 caacctggcc aagtatccag catccacagt gcagatcctt ggggctgaaa aggccctgtt
      661 cagagccctg aagacaaggg gtaacactcc aaaatatgga ctcattttcc actccacctt
      721 cattggccga gcagctgcca agaacaaagg ccgcatctcc cgatacctgg caaacaaatg
      781 cagtattgcc tcacgaatcg attgcttctc tgaggtgccc acgagtgtat tcggggagaa
      841 gcttcgagaa caagttgaag agcgactgtc cttctatgag actggacatc accatcacca
      901 tcaccattag gcggccgcct aggacccagc tttcttgtac aaagtggtaa gcttaattag
      961 ctgagcttgg actcctgttg atagatccag taatgacctc agaactccat ctggatttgt
     1021 tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagt aacaacacca
     1081 tttgtcgaga aatcataaaa aatttatttg ctttgtgagc ggataacaat tataatagat
     1141 tcaattgtga gcggataaca atttcacaca gaattcatta aagaggagaa attaactatg
     1201 ggatccttgg tgctgtttga aacgtctgtg ggttacgcca tctttaaggt tctaaatgag
     1261 aagaaacttc aagaggttga tagtttatgg aaagaatttg aaactccaga gaaagcaaac
     1321 aaaatagtaa agctaaaaca ttttgagaaa tttcaggata cagcagaagc attagcagca
     1381 ttcacagctc tgatggaggg caaaatcaat aagcagctga aaaaagttct gaagaaaata
     1441 gtaaaagaag cccatgaacc gctggcagta gctgatgcta aactaggagg ggtcataaag
     1501 gaaaagctga atctcagttg tatccatagt cctgttgtta atgaacttat gagaggaatt
     1561 cgttcacaaa tggatggatt aatccctggg gtagaaccac gtgaaatggc agctatgtgt
     1621 cttggattgg ctcacagcct gtctcgatat agattgaagt ttagcgctga taaagtagac
     1681 acaatgattg ttcaggcaat ttccttgtta gatgacttgg ataaagaact aaacaactac
     1741 attatgcgat gtagagaatg gtatggctgg catttccctg aattaggaaa aattatttca
     1801 gataatttaa catactgcaa gtgtttacag aaagttggcg ataggaagaa ctatgcctct
     1861 gccaagcttt ctgagttgct gccagaagaa gttgaagcag aagtgaaagc agctgcagag
     1921 atatcaatgg gaacagaggt ttcagaagaa gatatttgca atattctgca tctttgcacc
     1981 caggtgattg aaatctctga atatcgaacc cagctctatg aatatctaca aaatcgaatg
     2041 atggccattg cacccaatgt tacagtcatg gttggggaat tagttggagc acggcttatt
     2101 gctcatgcag gttctctttt aaatttggcc aagcatgcag cttctaccgt tcagattctt
     2161 ggagctgaaa aggcactttt cagagccctc aaatctagac gggatacccc taagtatggt
     2221 ctcatttatc atgcttcact cgtgggccag acaagtccca aacacaaagg aaagatttct
     2281 cgaatgctgg cagccaaaac cgttttggct atccgttatg atgcttttgg tgaggattca
     2341 agttctgcaa tgggagttga gaacagagcc aaattagagg ccaggttgag aactttggaa
     2401 gacagatagg cggccgccta ggacccagct ttcttgtaca aagtggtaag cttaattagc
     2461 tgagcttgga ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt
     2521 cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagta acaacaccat
     2581 ttgtcgagaa atcataaaaa atttatttgc tttgtgagcg gataacaatt ataatagatt
     2641 caattgtgag cggataacaa tttcacacag aattcattaa agaggagaaa ttaactatgg
     2701 gatctacaag tttgtacaaa aaagttggca ccatgactga ggctgatgtg aatccaaagg
     2761 cctatcccct tgccgatgcc cacctcacca agaagctact ggacctcgtt cagcagtcat
     2821 gtaactataa gcagcttcgg aaaggagcca atgaggccac caaaaccctc aacaggggca
     2881 tctctgagtt catcgtgatg gctgcagacg ccgagccact ggagatcatt ctgcacctgc
     2941 cgctgctgtg tgaagacaag aatgtgccct acgtgtttgt gcgctccaag caggccctgg
     3001 ggagagcctg tggggtctcc aggcctgtca tcgcctgttc tgtcaccatc aaagaaggct
     3061 cgcagctgaa acagcagatc caatccattc agcagtccat tgaaaggctc ttagtctaac
     3121 caactttctt gtacaaagtg gtaagcttaa ttagctgagc ttggactcct gttgatagat
     3181 ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg
     3241 ttttttattg gtgagaatcc aagtaacaac accatttaaa tggagtggtt acaaatggag
     3301 tggttaatta aggctagctt ggcgagattt tcaggagcta aggaagctaa aatggagaaa
     3361 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag
     3421 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc
     3481 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
     3541 gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg
     3601 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
     3661 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
     3721 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
     3781 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     3841 gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg
     3901 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga
     3961 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta atttttttaa
     4021 ggcagttatt ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat
     4081 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     4141 cgctctcctg agtaggacaa atccgccctc tagattacgt gcagtcgatg ataagctgtc
     4201 aaacatgaga attgtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     4261 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4321 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     4381 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     4441 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     4501 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     4561 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     4621 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     4681 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     4741 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     4801 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     4861 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     4921 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     4981 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     5041 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     5101 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     5161 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     5221 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     5281 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     5341 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgca
     5401 ccattcgatg gtgtcggaat ttcgggcagc gttgggtcct ggccacgggt gcgcatgatc
     5461 tagagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     5521 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     5581 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     5641 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccacatgc
     5701 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     5761 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5821 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5881 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5941 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6001 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6061 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6121 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6181 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6241 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6301 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6361 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6421 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6481 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6541 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6601 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6661 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6721 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6781 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6841 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6901 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6961 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7021 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7081 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7141 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7201 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7261 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7321 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7381 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7441 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7501 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7561 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7621 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7681 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     7741 ggccctttcg tcttcacctc gagcttaatt aagtcgcccc atcaaaaaaa tattctcaac
     7801 ataaaaaact ttgtgtaata cttgtaacgc tgaattcgcc cggatagctc agtcggtaga
     7861 gcagcggtca ctgtgatgat ggttttccaa cattcgcagt ttccaccaga aaggttttcc
     7921 ttagtgttgg gtaaaccttc cttggatgtc tgagtgaccg cgggtccagg gttcaagtcc
     7981 ctgttcgggc gccactgcag atccttagcg aaagctaagg atttttttta agcttaatta
     8041 gctgagcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
     8101 ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag taacaacacc
     8161 atttgtcgag aaatcataaa aaatttattt gctttgtgag cggataacaa ttataataga
     8221 ttcaattgtg agcggataac aatttcacac agaattcatt aaagaggaga aattaactat
     8281 gggatccggg aagaatgtga tggtggagcc gcatcggcat gagggtgtct tcatttgtcg
     8341 aggaaaggaa gatgcactgg tcaccaagaa cctggtccct ggggaatcag tttatggaga
     8401 gaagagagtc tcgatttcgg aaggagatga caaaattgag taccgagcct ggaacccctt
     8461 ccgctccaag ctagcagcag caatcctggg tggtgtggac cagatccaca tcaaaccggg
     8521 ggctaaggtt ctctacctcg gggctgcctc gggcaccacg gtctcccatg tctctgacat
     8581 cgttggtccg gatggtctag tctatgcagt cgagttctcc caccgctctg gccgtgacct
     8641 cattaacttg gccaagaaga ggaccaacat cattcctgtg atcgaggatg ctcgacaccc
     8701 acacaaatac cgcatgctca tcgcaatggt ggatgtgatc tttgctgatg tggcccagcc
     8761 agaccagacc cggattgtgg ccctgaatgc ccacaccttc ctgcgtaatg gaggacactt
     8821 tgtgatttcc attaaggcca actgcattga ctccacagcc tcagccgagg ccgtgtttgc
     8881 ctccgaagtg aaaaagatgc aacaggagaa catgaagccg caggagcagt tgacccttga
     8941 gccatatgaa agagaccatg ccgtggtcgt gggagtgtac aggccatagg cggccgccta
     9001 ggacccagct ttcttgtaca aagtggtaag cttaattagc tgagcttgga ctcctgttga
     9061 tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc
     9121 gggcgttttt tattggtgag aatccaagta acaacaccat ttgtcgagaa atcataaaaa
     9181 atttatttgc tttgtgagcg gataacaatt ataatagatt caattgtgag cggataacaa
     9241 tttcacacag aattcattaa agaggagaaa ttaactatgg gatctgtgct gttgcacgtg
     9301 ctgtttgagc acgcggtcgg ctacgcgctg ctggcgctga aggaagtgga ggagatcagt
     9361 ctgctgcagc cgcaggtgga ggagtccgtg ctcaacctgg gcaaattcca cagcatcgtt
     9421 cgtctggtgg ccttttgtcc ctttgcctca tcccaggttg ccttggaaaa tgccaacgcc
     9481 gtgtctgaag gggttgttca tgaggacctc cgcctgctct tggagaccca cctgccgtcc
     9541 aaaaagaaga aagtactctt gggagttggg gatcccaaga ttggtgccgc aatacaggag
     9601 gagttagggt acaactgcca gactggagg
//
