LOCUS       sequence-329902-       10523 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for a sgRNA against BFP and spCas9-Ec86 RT fusion 
            protein linked to mCherry via a T2A peptide. Donor template An was 
            inserted in the Retron Ec86 msr-msd region by SpeI and AvrII..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10523)
  AUTHORS   Zhao B, Chen SA, Lee J, Fraser HB
  TITLE     Bacterial Retrons Enable Precise Gene Editing in Human Cells.
  JOURNAL   CRISPR J. 2022 Jan 24. doi: 10.1089/crispr.2021.0065.
  PUBMED    35076284
REFERENCE   2  (bases 1 to 10523)
  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: "CRISPR J. 
            2022 Jan 24. doi: 10.1089/crispr.2021.0065."
COMMENT     Sequence Label: 
            pBZ210-pU6-sgBFP-An_CBh-sv40NLS-Cas9-Ec86RT-NLS-T2A-mCherry
FEATURES             Location/Qualifiers
     source          1..10523
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..254
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6..26
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     177..196
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        275..350
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        635..920
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        922..1199
                     /label=chicken beta-actin promoter
     intron          1200..1428
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1440..1449
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1449..1514
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1521..1541
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1566..5666
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1566..5666
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6723..6770
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6774..6827
                     /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             6834..7544
                     /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
                     EGRHSTGGMDELYK"
     primer_bind     complement(6979..6997)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7247..7266
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(7569..7586)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7575..7782
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7791..7931
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7791..7920
                     /label=AAV2 ITR
     rep_origin      8006..8461
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8093..8112)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8303..8324
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8478..8497)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8597..8619
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8657..8675)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8743..8847
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8848..8916,8917..9708)
                     /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     complement(9066..9085)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9879..10467
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10368..10387
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgtgaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
       61 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
      121 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
      181 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
      241 aaggacgaaa caccgctgaa gcactgcacg ccatgtttta gagctagaaa tagcaagtta
      301 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc cgtacgatgc
      361 gcacccttag cgagaggttt atcattaagg tcaacctctg gatgttgttt cggcatcctg
      421 cattgaatct gagttactgt ctgtttccct actagtaagt cgtgctgctt catgtggtcg
      481 gggtagcggc tgaagcactg cacgccgtac gtcagggtgg tcacgagggt gggccagggc
      541 acgggcagct tgccggtggt gcagatgaac ttcaggccta ggagggaacc cgtttcttct
      601 gacgtaaggg tgcgcatttt tttctagagg tacccgttac ataacttacg gtaaatggcc
      661 cgcctggctg accgcccaac gacccccgcc cattgacgtc aatagtaacg ccaataggga
      721 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      781 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      841 ggcattgtgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      901 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      961 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
     1021 tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg
     1081 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
     1141 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
     1201 gagtcgctgc gacgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
     1261 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
     1321 ccgggctgta attagctgag caagaggtaa gggtttaagg gatggttggt tggtggggta
     1381 ttaatgttta attacctgga gcacctgcct gaaatcactt tttttcaggt tggaccggtg
     1441 ccaccatgga ctataaggac cacgacggag actacaagga tcatgatatt gattacaaag
     1501 acgatgacga taagatggcc ccaaagaaga agcggaaggt cggtatccac ggagtcccag
     1561 cagccgacaa gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg
     1621 tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc
     1681 ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg
     1741 aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct
     1801 gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca
     1861 gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg
     1921 gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa
     1981 agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc
     2041 acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg
     2101 acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc
     2161 ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca
     2221 gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa
     2281 acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg
     2341 aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg
     2401 cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca
     2461 tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct
     2521 ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc
     2581 ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg
     2641 ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc
     2701 tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc
     2761 ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc
     2821 acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga
     2881 tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca
     2941 gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg
     3001 aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata
     3061 agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg
     3121 tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc
     3181 tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga
     3241 ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa
     3301 tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa
     3361 ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg
     3421 tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg
     3481 cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg
     3541 gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc
     3601 tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg
     3661 acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc
     3721 tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga
     3781 cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg
     3841 tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga
     3901 gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc
     3961 ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc
     4021 gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc
     4081 atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa
     4141 gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga
     4201 agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc
     4261 tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac
     4321 agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga
     4381 acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt
     4441 ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca
     4501 actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa
     4561 agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga
     4621 agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca
     4681 gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc
     4741 ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt
     4801 ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg
     4861 tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga
     4921 tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg
     4981 cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg
     5041 tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg
     5101 actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta
     5161 agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac
     5221 tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca
     5281 gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg
     5341 aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag
     5401 tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata
     5461 agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag
     5521 cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca
     5581 aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga
     5641 tcgacctgtc tcagctggga ggcgactctg gaggatctag cggaggatcc tctggcagcg
     5701 agacaccagg aacaagcgag tcagcaacac cagagagcag tggcggcagc agcggcggca
     5761 gcagcaagag tgcggaatac cttaacacat ttaggcttag aaacctggga ctccccgtta
     5821 tgaataacct gcacgatatg tctaaggcaa cccgaatcag tgtagagacg cttagactct
     5881 tgatatatac cgccgacttt cgatacagaa tatataccgt cgagaagaag ggcccagaga
     5941 aacgaatgcg gaccatatac caacctagta gggagctgaa agcgcttcag ggctgggtac
     6001 ttcgaaatat ccttgacaaa ctttccagtt caccgttcag cattggtttc gagaaacacc
     6061 agagtattct gaataacgcg acacctcaca taggagccaa tttcatcctc aatattgacc
     6121 tggaggattt cttccctagc cttactgcca ataaagtgtt cggtgtattc cacagcctcg
     6181 gctacaaccg actgatttca tctgtactca caaaaatttg ttgctataag aacctgctcc
     6241 ctcaaggggc cccaagtagc ccaaaactgg caaacctcat ctgttcaaaa ttggactatc
     6301 gaattcaggg ctatgccgga tccaggggct tgatctacac gagatacgca gacgacctga
     6361 cactttcagc acaatccatg aagaaagttg ttaaagcgag ggattttctt ttttccatta
     6421 ttccgtctga aggattggtt attaattcta aaaaaacttg cattagtggt cctcggtctc
     6481 aacgaaaggt tacaggcctg gtaatctctc aggagaaggt tgggattgga agagagaagt
     6541 ataaagagat tcgcgccaaa atacatcata ttttttgcgg taaatcatct gaaatcgagc
     6601 acgtaagggg atggctttct ttcattcttt ctgtagactc caaatctcac cgacgactca
     6661 tcacatatat aagtaaactg gagaaaaaat atgggaaaaa tccgcttaac aaggctaaaa
     6721 ctaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag cttgagggca
     6781 gaggaagtct gctaacatgc ggtgacgtgg aggagaatcc cggccctgct agcatggtga
     6841 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     6901 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     6961 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     7021 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     7081 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     7141 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     7201 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     7261 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     7321 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     7381 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     7441 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     7501 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtgagaattc ctagagctcg
     7561 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     7621 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     7681 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     7741 caagggggag gattgggaag agaatagcag gcatgctggg gagcggccgc aggaacccct
     7801 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     7861 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     7921 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     7981 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     8041 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     8101 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     8161 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     8221 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     8281 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     8341 atctcgggct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     8401 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     8461 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     8521 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     8581 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     8641 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     8701 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     8761 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     8821 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     8881 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     8941 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     9001 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     9061 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     9121 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     9181 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     9241 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     9301 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     9361 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     9421 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     9481 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     9541 aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg gccagatggt
     9601 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     9661 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     9721 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     9781 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     9841 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     9901 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     9961 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
    10021 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
    10081 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
    10141 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
    10201 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
    10261 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
    10321 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
    10381 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
    10441 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
    10501 gccttttgct ggccttttgc tca
//
