LOCUS       sequence-488946-        8473 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  Recombinant protein expression of PCMTD1 F57 F217 F220.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8473)
  AUTHORS   Pang EZ, Zhao B, Flowers C, Oroudjeva E, Winter JB, Pandey V, Sawaya
            MR, Wohlschlegel J, Loo JA, Rodriguez JA, Clarke SG
  TITLE     Structural basis for L-isoaspartyl-containing protein recognition by
            the PCMTD1 cullin-RING E3 ubiquitin ligase.
  JOURNAL   bioRxiv [Preprint]. 2025 May 21:2025.05.21.654933. doi: 
            10.1101/2025.05.21.654933.
  PUBMED    40475564
REFERENCE   2  (bases 1 to 8473)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 9, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2025 May 21:2025.05.21.654933. doi: 
            10.1101/2025.05.21.654933."
COMMENT     Sequence Label: pMAPLe4 PCMTD1 F57 F217 F220
FEATURES             Location/Qualifiers
     source          1..8473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1117..1135)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1131..1178
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1215..1670
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1302..1321)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1512..1533
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1762..2577)
                     /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(1762..2577)
                     /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     2485..2504
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2569..2588
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2699..3287
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3188..3207
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3441..3458
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3473..3612
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3538..3557)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3698..3720
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3714..3905)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4648..4666
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(4687..4705,4706..4711,4712..4728,4729..4734,
                     4735..4760)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    4687..4705
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4712..4730
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             4770..4778
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             4784..5494
                     /codon_start=1
                     /product="tobacco vein mottling virus NIa protease 
                     (Nallamsetty et al., 2004)"
                     /label=TVMV protease
                     /translation="MSKALLKGVRDFNPISACVCLLENSSDGHSERLFGIGFGPYIIAN
                     QHLFRRNNGELTIKTMHGEFKVKNSTQLQMKPVEGRDIIVIKMAKDFPPFPQKLKFRQP
                     TIKDRVCMVSTNFQQKSVSSLVSESSHIVHKEDTSFWQHWITTKDGQCGSPLVSIIDGN
                     ILGIHSLTHTTNGSNYFVEFPEKFVATYLDAADGWCKNWKFNADKISWGSFTLVEDAPE
                     DDFMAKKTVAAIMD"
     terminator      5527..5613
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5739..6821)
                     /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     6783..6802
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6822..6899)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7105..7124
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     7208..7227
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7208..7226
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7227..7251
                     /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             7266..7288
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             7295..8395
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      8252..8261
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     8369..8392
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             8405..8425
                     /codon_start=1
                     /product="tobacco vein mottling virus (TVMV) NIa protease 
                     recognition and cleavage site
                     "
                     /label=TVMV site
                     /translation="ETVRFQS"
     CDS             8426..8443
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             8453..8473
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
ORIGIN
        1 atgggtgggg ctgtctcggc gggagaagac aacgatgatc tgattgataa cttaaaggaa
       61 gcacaatata ttcgtacaga acgggttgaa caagcattca gagcaattga tcgcggggat
      121 tattatttag aaggttatcg cgataacgcc tacaaagatc tggcctggtt tcacggcaac
      181 attcatttga gcgccccgtg tatctatagc gaggtaatgg aggcgctgaa acttcagcct
      241 gggctctcat ttttgaacct gggatccggt accggctacc tctcaacgat ggtcggactg
      301 attcttggtc cgtttggtat caaccacggc attgaattac atagcgatgt tgtggagtat
      361 gctaaagaga agctggaaag ttttatcaaa aactctgact cctttgataa atttgaattt
      421 tgtgaaccgg cttttgtggt cggaaactgt ttacagatcg cgtcagatag ccatcagtat
      481 gatcgcatct attgcggagc aggtgtccag aaagatcacg aaaactatat gaaaattctg
      541 cttaaagtcg gtgggatact tgtgatgccg atcgaagatc aactgacgca aattatgcgc
      601 actggccaaa acacgtggga aagcaagaat attttggcgg tgtccttctt tccgctgttt
      661 cagcccagca agaacgacaa cggaaaaccg gattctgttg gtttacctcc gtgtgcggtg
      721 agaaatttac aggatcttgc acggatttat attcgccgca ccttgcgtaa tttcataaat
      781 gacgaaatgc aagccaaagg tattccacaa cgtgcaccac ctaaacgaaa acggaaacgg
      841 gttaagcaac gcattaatac atatgtattt gttggtaatc agttgatccc acagccgtta
      901 gattctgagg aggatgaaaa aatggaggaa gacataaagg aggaagagga aaaagatcat
      961 aatgaagcca tgaaaccgga agaacctcca caaaacctgt tacgagaaaa aattatgaag
     1021 cttccgctgc ccgagtcact taaagcctat ttgacgtatt tccgggataa ataactgcta
     1081 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     1141 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
     1201 gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     1261 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     1321 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     1381 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     1441 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     1501 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     1561 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     1621 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     1681 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     1741 tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     1801 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     1861 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     1921 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     1981 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     2041 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     2101 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     2161 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     2221 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     2281 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     2341 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     2401 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     2461 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     2521 catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac
     2581 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt
     2641 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2701 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2761 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2821 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     2881 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2941 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3001 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3061 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3121 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3181 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3241 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3301 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3361 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3421 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     3481 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
     3541 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     3601 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     3661 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     3721 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     3781 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     3841 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     3901 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     3961 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     4021 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     4081 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     4141 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
     4201 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
     4261 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
     4321 cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc
     4381 atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag
     4441 gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg
     4501 ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg
     4561 agttgcatga taaagaagac agtcataagt gcggctaaga tagtcgacgt ctaagaaacc
     4621 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcac
     4681 ctcgagtccc tatcagtgat agagattgac atccctatca gtgatagaga tactgagcac
     4741 atcagcagga cgcactgacc gaattgattt aaggaggtaa catatgtcta aagctttgct
     4801 gaagggcgtg cgcgatttta atccgatctc tgcttgcgta tgcctgctgg aaaactcctc
     4861 ggatggtcat agtgaacgtc tgtttggcat tggttttggc ccgtatatca ttgccaacca
     4921 gcatctgttt cgtcgtaaca atggcgaact gaccatcaaa accatgcatg gtgaattcaa
     4981 agtcaaaaac tctacccagc tgcagatgaa accggttgaa ggccgtgaca ttatcgttat
     5041 caaaatggct aaagacttcc cgccgttccc gcagaaactg aaattccgtc agccgaccat
     5101 caaagatcgt gtgtgcatgg tgtccaccaa ctttcagcag aaaagcgtct cgagcctggt
     5161 gtctgaatcc tctcacattg tgcataaaga agacacttct ttctggcagc actggatcac
     5221 cactaaagat ggccagtgtg gcagcccact agtttccatc attgatggca acattctggg
     5281 catccacagc ctgactcata ccaccaacgg tagcaactac ttcgtggaat ttccggaaaa
     5341 attcgtggcg acttatctag atgccgcgga tggttggtgc aaaaactgga aattcaacgc
     5401 ggataaaatc agctggggtt cctttaccct ggttgaagat gcgccggaag atgacttcat
     5461 ggccaaaaaa actgttgccg ccatcatgga ctaataaaga tcccatggta cgcgtgctag
     5521 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     5581 ttgtcggtga acgctctcct gagtaggaca aatccgccgc cctagtagtc atgccccgcg
     5641 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     5701 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5761 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5821 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     5881 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     5941 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     6001 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
     6061 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     6121 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     6181 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     6241 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     6301 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     6361 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     6421 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     6481 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     6541 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     6601 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     6661 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     6721 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     6781 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     6841 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
     6901 gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     6961 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     7021 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     7081 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     7141 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc
     7201 gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa
     7261 ataattttgt ttaactttaa gaaggagata taccatgaaa atcgaagaag gtaaactggt
     7321 aatctggatt aacggcgata aaggctataa cggtctcgct gaagtcggta agaaattcga
     7381 gaaagatacc ggaattaaag tcaccgttga gcatccggat aaactggaag agaaattccc
     7441 acaggttgcg gcaactggcg atggccctga cattatcttc tgggcacacg accgctttgg
     7501 tggctacgct caatctggcc tgttggctga aatcaccccg gacaaagcgt tccaggacaa
     7561 gctgtatccg tttacctggg atgccgtacg ttacaacggc aagctgattg cttacccgat
     7621 cgctgttgaa gcgttatcgc tgatttataa caaagatctg ctgccgaacc cgccaaaaac
     7681 ctgggaagag atcccggcgc tggataaaga actgaaagcg aaaggtaaga gcgcgctgat
     7741 gttcaacctg caagaaccgt acttcacctg gccgctgatt gctgctgacg ggggttatgc
     7801 gttcaagtat gaaaacggca agtacgacat taaagacgtg ggcgtggata acgctggcgc
     7861 gaaagcgggt ctgaccttcc tggttgacct gattaaaaac aaacacatga atgcagacac
     7921 cgattactcc atcgcagaag ctgcctttaa taaaggcgaa acagcgatga ccatcaacgg
     7981 cccgtgggca tggtccaaca tcgacaccag caaagtgaat tatggtgtaa cggtactgcc
     8041 gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg ctgagcgcag gtattaacgc
     8101 cgccagtccg aacaaagagc tggcaaaaga gttcctcgaa aactatctgc tgactgatga
     8161 aggtctggaa gcggttaata aagacaaacc gctgggtgcc gtagcgctga agtcttacga
     8221 ggaagagttg gcgaaagatc cacgtattgc cgccaccatg gaaaacgccc agaaaggtga
     8281 aatcatgccg aacatcccgc agatgtccgc tttctggtat gccgtgcgta ctgcggtgat
     8341 caacgccgcc agcggtcgtc agactgtcga tgaagccctg aaagacgcgc agactaatgg
     8401 ctcagaaaca gttcggttcc agtctcatca ccatcaccat cactcgagct cagagaacct
     8461 gtatttccag agt
//
