LOCUS       sequence-340901-       11247 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses spNW100 in Bl21.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11247)
  AUTHORS   Zhang S, Ren Q, Novick SJ, Strutzenberg TS, Griffin PR, Bao H
  TITLE     One-step construction of circularized nanodiscs using 
            SpyCatcher-SpyTag.
  JOURNAL   Nat Commun. 2021 Sep 14;12(1):5451. doi: 10.1038/s41467-021-25737-7.
  PUBMED    34521837
REFERENCE   2  (bases 1 to 11247)
  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: "Nat 
            Commun."; date: "2021-09-14"; volume: "12(1):5451. doi"; pages: " 
            10.1038/s41467-021-25737-7"
COMMENT     Sequence Label: spNW100
FEATURES             Location/Qualifiers
     source          1..11247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(99..118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     309..330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(559..1374)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(559..1374)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1282..1301
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1366..1385
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1496..2084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1985..2004
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2238..2255
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2270..2409
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2335..2354)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2495..2517
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2511..2702)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3511..4593)
                     /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     4555..4574
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4594..4671)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4877..4896
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4980..4999
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4980..4998
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4999..5023
                     /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             5038..5060
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5079..5096
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5106..5123
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5127..5159
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             5178..5516
                     /codon_start=1
                     /product="improved SpyCatcher variant that shows rapid 
                     reaction with SpyTag002 (Keeble et al., 2017)"
                     /label=SpyCatcher002
                     /translation="VTTLSGLSGEQGPSGDMTTEEDSATHIKFSKRDEDGRELAGATME
                     LRDSSGKTISTWISDGHVKDFYLYPGKYTFVETAAPDGYEVATAITFTVNEQGQVTVNG
                     EATKGDAHT"
     CDS             11031..11072
                     /codon_start=1
                     /product="improved SpyTag variant that shows rapid reaction
                     with SpyCatcher002 (Keeble et al., 2017)"
                     /label=SpyTag002
                     /translation="VPTIVMVDAYKRYK"
     CDS             11091..11108
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(11161..11179)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      11175..11222
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
      481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      541 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
      601 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
      661 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
      721 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
      781 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
      841 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
      901 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
      961 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     1021 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     1081 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     1141 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     1201 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     1261 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     1321 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcataacacc
     1381 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgaccaaa atcccttaac
     1441 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1501 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1561 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1621 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     1681 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1741 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1801 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1861 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     1921 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     1981 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2041 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2101 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2161 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     2221 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     2281 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
     2341 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     2401 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     2461 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     2521 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     2581 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     2641 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     2701 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     2761 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     2821 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     2881 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     2941 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     3001 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     3061 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     3121 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg
     3181 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     3241 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     3301 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     3361 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     3421 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     3481 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3541 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3601 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     3661 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     3721 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     3781 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     3841 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     3901 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     3961 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     4021 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     4081 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     4141 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     4201 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     4261 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     4321 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     4381 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     4441 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     4501 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     4561 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     4621 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     4681 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     4741 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     4801 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     4861 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     4921 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
     4981 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
     5041 gtttaacttt aagaaggaga tataccatgg gcagcagcca tcatcatcat catcacagca
     5101 gcggcctggt gccgcgcggc agccatatgg ctagcatgac tggtggacag caaatgggtc
     5161 gcggatccgg cgccatggta accaccttat caggtttatc aggtgagcaa ggtccgtccg
     5221 gtgatatgac aactgaagaa gatagtgcta cccatattaa attctcaaaa cgtgatgagg
     5281 acggccgtga gttagctggt gcaactatgg agttgcgtga ttcatctggt aaaactatta
     5341 gtacatggat ttcagatgga catgtgaagg atttctacct gtatccagga aaatatacat
     5401 ttgtcgaaac cgcagcacca gacggttatg aggtagcaac tgctattacc tttacagtta
     5461 atgagcaagg tcaggttact gtaaatggcg aagcaactaa aggtgacgct catactagta
     5521 ccttctccaa actgcgcgaa cagctgggtc cggtgacgca agaattctgg gataacctgg
     5581 aaaaagaaac cgaaggcctg cgtcaggaaa tgagtaaaga cctggaagaa gtgaaagcca
     5641 aagttcaacc gtatctggat gacttccaga aaaaatggca agaagaaatg gaactgtacc
     5701 gccagaaagt cgaaccgctg cgtgccgaac tgcaggaagg cgcacgccaa aaactgcatg
     5761 aactgcaaga aaaactgagc ccgctgggtg aagaaatgcg tgatcgcgcc cgtgcacatg
     5821 ttgatgctct gcgtacccac ctggccccgt attctgatga actgcgtcag cgtctggccg
     5881 cacgtctgga agcgctgaaa gaaaacggcg gtgcccgtct ggcagaatac catgctaaag
     5941 cgaccgaaca cctgtcaacg ctgtcggaaa aagctaaacc ggccctggaa gatctgcgcc
     6001 agggtctgct gccggttctg gaatcattta aagtcagctt cctgtctgcc ctggaagaat
     6061 atacgaaaaa actgaacacc cagggcacgc cggttaccca ggaattttgg gacaatctgg
     6121 aaaaagaaac ggaaggcctg cgccaggaaa tgtcgaaaga cctggaagaa gtcaaagcaa
     6181 aagtgcagcc gtacctggac gattttcaga aaaaatggca agaagaaatg gagctgtacc
     6241 gccagaaagt tgaaccgctg cgcgcggaac tgcaggaagg cgcccgtcag aaactgcacg
     6301 aactgcagga aaaactgagt ccgctgggcg aagaaatgcg tgatcgtgcc cgcgcacacg
     6361 ttgatgccct gcgcacgcac ctggccccgt attccgatga actgcgccag cgcctggcag
     6421 ctcgtctgga agccctgaaa gaaaatggcg gtgcccgcct ggccgaatat cacgccaaag
     6481 ccaccgaaca cctgagtacg ctgtccgaaa aagctaaacc ggcgctggaa gatctgcgcc
     6541 aaggtctgct gccggtgctg gaaagcttta aagtttcatt cctgtcggcg ctggaagaat
     6601 acacgaaaaa actgaacacg cagggcaccc cggttaccca ggaattctgg gacaacctgg
     6661 aaaaggagac ggaaggcctg cgtcaagaaa tgagcaaaga tctggaagaa gttaaagcca
     6721 aagtccagcc gtacctggac gactttcaga aaaaatggca agaagaaatg gagttatacc
     6781 gccagaaagt ggaaccgctg cgcgctgaac tgcaggaagg cgcgcgtcag aaactgcatg
     6841 agttacagga aaaactgtca ccgctgggag aggaaatgcg cgaccgtgcc cgtgcacacg
     6901 ttgacgccct gcgcacccac ctggcgccgt attcggacga actgcgtcaa cgcctggccg
     6961 cccgcctgga agccttaaag gaaaatggcg gtgcccgtct ggccgaatat cacgcgaaag
     7021 ccacggaaca tctgagcacg ctgtctgaaa aagccaagcc tgccctggaa gatctgcgcc
     7081 aggggctgct gccggtcctg gaaagtttta aagttagttt cctgtccgca ctggaagagt
     7141 atacgaaaaa actgaatacc cagggcacgc cggtgaccca ggaattttgg gataatctgg
     7201 aaaaggagac cgaaggcctg cgccaagaaa tgtccaagga tctggaagaa gtgaaagcga
     7261 aagtgcagcc gtatctggac gatttccaga aaaaatggca agaagaaatg gagctatatc
     7321 gtcaaaaagt tgaaccgctg cgtgcggagt tacaggaagg cgcccgtcaa aaactgcacg
     7381 agttacaaga gaagctgagc ccgctgggcg aggagatgcg cgatcgtgcc cgcgcacatg
     7441 ttgacgccct gcgtacgcac ctggcaccgt acagcgacga actgcgccaa cgcctggccg
     7501 cccgactgga agccttaaaa gagaatggcg gtgcccgatt agccgaatat cacgcaaaag
     7561 ctacggaaca tctgtctacc ctgagcgaaa aagccaagcc cgccctggaa gatctgcgcc
     7621 agggcttatt accggtcctg gaaagcttca aagttagctt cctgagcgcg ctggaagaat
     7681 ataccaaaaa actgaatacc caaggcaccc cggttaccca agaattttgg gacaacctgg
     7741 aaaaagagac ggagggcctg cgccaagaaa tgagcaagga cctggaagaa gtcaaggcga
     7801 aagtgcaacc gtacctggac gacttccaga aaaaatggca agaagaaatg gagctttatc
     7861 gtcaaaaagt cgaaccgctg cgcgccgagt tacaggaagg cgcgcgtcaa aaactgcacg
     7921 aattacaaga gaaactgagt ccgctgggag aggagatgcg cgaccgcgcc cgtgcacacg
     7981 tggacgcact gcgtacgcat ctggcgccgt acagcgatga gttacgtcaa cgcctggccg
     8041 cgcgcctgga agctttaaag gagaacggcg gtgcccgcct ggcggaatat cacgccaaag
     8101 ctacggaaca cctgagcacc ctgagcgaaa aagcgaagcc agccctggaa gatctgcgcc
     8161 agggattatt gccggtcctg gaatctttca aagtgtcatt cctgagcgcc ccggaagaat
     8221 acaccaaaaa actgaacacc caactgccgg gtaccggtgc agctgccctg gaaagtacct
     8281 tctccaaact gcgcgaacag ctgggtccgg tgacgcaaga attctgggat aacctggaaa
     8341 aagaaaccga aggcctgcgt caggaaatga gtaaagacct ggaagaagtg aaagccaaag
     8401 ttcaaccgta tctggatgac ttccagaaaa aatggcaaga agaaatggaa ctgtaccgcc
     8461 agaaagtcga accgctgcgt gccgaactgc aggaaggcgc acgccaaaaa ctgcatgaac
     8521 tgcaagaaaa actgagcccg ctgggtgaag aaatgcgtga tcgcgcccgt gcacatgttg
     8581 atgctctgcg tacccacctg gccccgtatt ctgatgaact gcgtcagcgt ctggccgcac
     8641 gtctggaagc gctgaaagaa aacggcggtg cccgtctggc agaataccat gctaaagcga
     8701 ccgaacacct gtcaacgctg tcggaaaaag ctaaaccggc cctggaagat ctgcgccagg
     8761 gtctgctgcc ggttctggaa tcatttaaag tcagcttcct gtctgccctg gaagaatata
     8821 cgaaaaaact gaacacccag ggcacgccgg ttacccagga attttgggac aatctggaaa
     8881 aagaaacgga aggcctgcgc caggaaatgt cgaaagacct ggaagaagtc aaagcaaaag
     8941 tgcagccgta cctggacgat tttcagaaaa aatggcaaga agaaatggag ctgtaccgcc
     9001 agaaagttga accgctgcgc gcggaactgc aggaaggcgc ccgtcagaaa ctgcacgaac
     9061 tgcaggaaaa actgagtccg ctgggcgaag aaatgcgtga tcgtgcccgc gcacacgttg
     9121 atgccctgcg cacgcacctg gccccgtatt ccgatgaact gcgccagcgc ctggcagctc
     9181 gtctggaagc cctgaaagaa aatggcggtg cccgcctggc cgaatatcac gccaaagcca
     9241 ccgaacacct gagtacgctg tccgaaaaag ctaaaccggc gctggaagat ctgcgccaag
     9301 gtctgctgcc ggtgctggaa agctttaaag tttcattcct gtcggcgctg gaagaataca
     9361 cgaaaaaact gaacacgcag ggcaccccgg ttacccagga attctgggac aacctggaaa
     9421 aggagacgga aggcctgcgt caagaaatga gcaaagatct ggaagaagtt aaagccaaag
     9481 tccagccgta cctggacgac tttcagaaaa aatggcaaga agaaatggag ttataccgcc
     9541 agaaagtgga accgctgcgc gctgaactgc aggaaggcgc gcgtcagaaa ctgcatgagt
     9601 tacaggaaaa actgtcaccg ctgggagagg aaatgcgcga ccgtgcccgt gcacacgttg
     9661 acgccctgcg cacccacctg gcgccgtatt cggacgaact gcgtcaacgc ctggccgccc
     9721 gcctggaagc cttaaaggaa aatggcggtg cccgtctggc cgaatatcac gcgaaagcca
     9781 cggaacatct gagcacgctg tctgaaaaag ccaagcctgc cctggaagat ctgcgccagg
     9841 ggctgctgcc ggtcctggaa agttttaaag ttagtttcct gtccgcactg gaagagtata
     9901 cgaaaaaact gaatacccag ggcacgccgg tgacccagga attttgggat aatctggaaa
     9961 aggagaccga aggcctgcgc caagaaatgt ccaaggatct ggaagaagtg aaagcgaaag
    10021 tgcagccgta tctggacgat ttccagaaaa aatggcaaga agaaatggag ctatatcgtc
    10081 aaaaagttga accgctgcgt gcggagttac aggaaggcgc ccgtcaaaaa ctgcacgagt
    10141 tacaagagaa gctgagcccg ctgggcgagg agatgcgcga tcgtgcccgc gcacatgttg
    10201 acgccctgcg tacgcacctg gcaccgtaca gcgacgaact gcgccaacgc ctggccgccc
    10261 gactggaagc cttaaaagag aatggcggtg cccgattagc cgaatatcac gcaaaagcta
    10321 cggaacatct gtctaccctg agcgaaaaag ccaagcccgc cctggaagat ctgcgccagg
    10381 gcttattacc ggtcctggaa agcttcaaag ttagcttcct gagcgcgctg gaagaatata
    10441 ccaaaaaact gaatacccaa ggcaccccgg ttacccaaga attttgggac aacctggaaa
    10501 aagagacgga gggcctgcgc caagaaatga gcaaggacct ggaagaagtc aaggcgaaag
    10561 tgcaaccgta cctggacgac ttccagaaaa aatggcaaga agaaatggag ctttatcgtc
    10621 aaaaagtcga accgctgcgc gccgagttac aggaaggcgc gcgtcaaaaa ctgcacgaat
    10681 tacaagagaa actgagtccg ctgggagagg agatgcgcga ccgcgcccgt gcacacgtgg
    10741 acgcactgcg tacgcatctg gcgccgtaca gcgatgagtt acgtcaacgc ctggccgcgc
    10801 gcctggaagc tttaaaggag aacggcggtg cccgcctggc ggaatatcac gccaaagcta
    10861 cggaacacct gagcaccctg agcgaaaaag cgaagccagc cctggaagat ctgcgccagg
    10921 gattattgcc ggtcctggaa tctttcaaag tgtcattcct gagcgccctg gaagaataca
    10981 ccaaaaaact gaacacccaa ctgccgggta ccggtgcagc tgccctggaa gtgcctacta
    11041 tcgtgatggt ggacgcctac aagcgttaca agtaagcggc cgcactcgag caccaccacc
    11101 accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
    11161 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
    11221 tgctgaaagg aggaactata tccggat
//
