LOCUS       sequence-226673-       13406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pET-28b backbone plasmid, Level 2 Golden Gate expression plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13406)
  AUTHORS   Pullmann P, Ulpinnis C, Marillonnet S, Gruetzner R, Neumann S, 
            Weissenborn MJ
  TITLE     Golden Mutagenesis: An efficient multi-site-saturation mutagenesis 
            approach by Golden Gate cloning with automated primer design.
  JOURNAL   Sci Rep. 2019 Jul 29;9(1):10932. doi: 10.1038/s41598-019-47376-1.
  PUBMED    31358887
REFERENCE   2  (bases 1 to 13406)
  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: "Sci Rep."; 
            date: "2019-07-29"; volume: "9(1):10932. doi"; pages: " 
            10.1038/s41598-019-47376-1"
COMMENT     Sequence Label: pAGM22082_cRed
FEATURES             Location/Qualifiers
     source          1..13406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     705..724
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(744..821)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1027..1046
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1130..1149
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1130..1148
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1149..1173
                     /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)."
     CDS             1231..1248
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1258..1275
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1279..1311
                     /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"
     regulatory      7433..7442
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(9470..9488)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      9484..9531
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      9568..10023
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9655..9674)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9865..9886
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(10115..10930)
                     /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(10115..10930)
                     /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     10838..10857
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     10922..10941
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      11052..11640
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11541..11560
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11794..11811
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    11826..11965
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(11891..11910)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12051..12073
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(12067..12258)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(13067..13406,1..743))
                     /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"
ORIGIN
        1 tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt
       61 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
      121 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
      181 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
      241 cgcgtaccgt cctcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
      301 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
      361 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
      421 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
      481 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
      541 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
      601 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
      661 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
      721 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
      781 catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc
      841 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
      901 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
      961 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     1021 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     1081 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc
     1141 actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt gcctttaact
     1201 ttaagaagga gatataccat gggcagcagc catcatcatc atcatcacag cagcggcctg
     1261 gtgccgcgcg gcagccatat ggctagcatg actggtggac agcaaatggg tcgggatgga
     1321 atgtgagacc agtggtatgg ggtaccgcac ggtctgccaa tccgacggag gtttatgaat
     1381 tttccacctt ttccacaagc tcaactagta ttaacgatgt ggatttagca aaaaaaacct
     1441 gtaaccctaa atgtaaaata acgggtaagc ctgccaacca tgttatggca gattaagcgt
     1501 ctttttgaag ggcaccgcat ctttcgcgtt gccgtaaatg tatccgttta gcaataagga
     1561 aattcttaat gaataatccg tcgttactca atcatgcggt cgaaacgatg gcagttggct
     1621 cgaaaagttt tgcgacagcc tcaaagttat ttgatgcaaa aacccggcgc agcgtactga
     1681 tgctctacgc ctggtgccgc cattgtgacg atgttattga cgatcagacg ctgggctttc
     1741 aggcccggca gcctgcctta caaacgcccg aacaacgtct gatgcaactt gagatgaaaa
     1801 cgcgccaggc ctatgcagga tcgcagatgc acgaaccggc gtttgcggct tttcgggaag
     1861 tggctatggc tcatgatatc gccccggctt acgcgtttga tcatctggaa ggcttcgcca
     1921 tggatgtacg cgaagcgcaa tacagccaac tggatgatac gctgcgctat tgctatcacg
     1981 ttgcaggcgt tgtcggcttg atgatggcgc aaatcatggg cgtgcgggat aacgccacgc
     2041 tggaccgcgc ctgtgacctt gggctggcat ttcagttgac caatattgct cgcgatattg
     2101 tggacgatgc gcatgcgggc cgctgttatc tgccggcaag ctggctggag catgaaggtc
     2161 tgaacaaaga gaattatgcg gcacctgaaa accgtcaggc gctgagccgt atcgcccgtc
     2221 gtttggtgca ggaagcagaa ccttactatt tgtctgccac agccggcctg gcagggttgc
     2281 ccctgcgttc cgcctgggca atcgctacgg cgaagcaggt ttaccggaaa ataggtgtca
     2341 aagttgaaca ggccggtcag caagcctggg atcagcggca gtcaacgacc acgcccgaaa
     2401 aattaacgct gctgctggcc gcctctggtc aggcccttac ttcccggatg cgggctcatc
     2461 ctccccgccc tgcgcatctc tggcagcgcc cgctctaggc ttactataag gatgtttgca
     2521 atgaaaccaa ctacggtaat tggtgcaggc ttcggtggcc tggcactggc aattcgtcta
     2581 caagctgcgg ggatccccgt cttactgctt gaacaacgtg ataaacccgg cggtcgggct
     2641 tatgtctacg aggatcaggg gtttaccttt gatgcaggcc cgacggttat caccgatccc
     2701 agtgccattg aagaactgtt tgcactggca ggaaaacagt taaaagagta tgtcgaactg
     2761 ctgccggtta cgccgtttta ccgcctgtgt tgggagtcag ggaaggtctt taattacgat
     2821 aacgatcaaa cccggctcga agcgcagatt cagcagttta atccccgcga tgtcgaaggt
     2881 tatcgtcagt ttctggacta ttcacgcgcg gtgtttaaag aaggctatct aaagctcggt
     2941 actgtccctt ttttatcgtt cagagacatg cttcgcgccg cacctcaact ggcgaaactg
     3001 caggcatgga gaagcgttta cagtaaggtt gccagttaca tcgaagatga acatctgcgc
     3061 caggcgtttt ctttccactc gctgttggtg ggcggcaatc ccttcgccac ctcatccatt
     3121 tatacgttga tacacgcgct ggagcgtgag tggggcgtct ggtttccgcg tggcggcacc
     3181 ggcgcattag ttcaggggat gataaagctg tttcaggatc tgggtggcga agtcgtgtta
     3241 aacgccagag tcagccatat ggaaacgaca ggaaacaaga ttgaagccgt gcatttagag
     3301 gacggtcgca ggttcctgac gcaagccgtc gcgtcaaatg cagatgtggt tcatacctat
     3361 cgcgacctgt taagccagca ccctgccgcg gttaagcagt ccaacaaact gcagactaag
     3421 cgcatgagta actctctgtt tgtgctctat tttggtttga atcaccatca tgatcagctc
     3481 gcgcatcaca cggtttgttt cggcccgcgt taccgcgagc tgattgacga aatttttaat
     3541 catgatggcc tcgcagagga cttctcactt tatctgcacg cgccctgtgt cacggattcg
     3601 tcactggcgc ctgaaggttg cggcagttac tatgtgttgg cgccggtgcc gcatttaggc
     3661 accgcgaacc tcgactggac ggttgagggg ccaaaactac gcgaccgtat ttttgcgtac
     3721 cttgagcagc attacatgcc tggcttacgg agtcagctgg tcacgcaccg gatgtttacg
     3781 ccgtttgatt ttcgcgacca gcttaatgcc tatcatggct cagccttttc tgtggagccc
     3841 gttcttaccc agagcgcctg gtttcggccg cataaccgcg ataaaaccat tactaatctc
     3901 tacctggtcg gcgcaggcac gcatcccggc gcaggcattc ctggcgtcat cggctcggca
     3961 aaagcgacag caggtttgat gctggaggat ctgatatgag cttttaccca ggagacagag
     4021 aatgcaaccg cattatgatc tgattctcgt gggggctgga ctcgcgaatg gccttatcgc
     4081 cctgcgactt cagcagcagc aacctgatat gcgtattttg cttatcgacg ccgcacccca
     4141 ggcgggcggg aatcatacgt ggtcatttca ccacgatgat ttgactgaga gccaacatcg
     4201 ttggatagct ccgctggtgg ttcatcactg gcccgactat caggtacgct ttcccacacg
     4261 ccgtcgtaag ctgaacagcg gctacttttg tattacttct cagcgtttcg ctgaggtttt
     4321 acagcgacag tttggcccgc acttgtggat ggataccgcg gtcgcagagg ttaatgcgga
     4381 atctgttcgg ttgaaaaagg gtcaggttat cggtgcccgc gcggtgattg acgggcgggg
     4441 ttatgcggca aattcagcac tgagcgtggg cttccaggcg tttattggcc aggaatggcg
     4501 attgagtcac ccgcatggtt tatcatctcc gattatcatg gatgccacgg tcgatcagca
     4561 aaatggttat cgcttcgtgt acagcctgcc gctctcgccg accagattgt taattgaaga
     4621 tacgcactat attgataatg cgacattaga tcctgaatgc gcgcggcaaa atatttgcga
     4681 ctatgccgcg caacagggtt ggcagcttca gacactgctg cgagaagaac agggcgcctt
     4741 acccattact ctgtcgggca atgccgacgc attctggcag cagcgccccc tggcctgtag
     4801 tggattacgt gccggtctgt tccatcctac caccggctat tcactgccgc tggcggttgc
     4861 cgtggccgac cgcctgagtg cacttgatgt ctttacgtcg gcctcaattc accatgccat
     4921 tacgcatttt gcccgcgagc gctggcagca gcagggcttt ttccgcatgc tgaatcgcat
     4981 gctgttttta gccggacccg ccgattcacg ctggcgggtt atgcagcgtt tttatggttt
     5041 acctgaagat ttaattgccc gtttttatgc gggaaaactc acgctgaccg atcggctacg
     5101 tattctgagc ggcaagccgc ctgttccggt attagcagca ttgcaagcca ttatgacgac
     5161 tcatcgttaa gcttcagagg aggaattaag taatgagttt tgatattgcc aaatacccga
     5221 ccctggcact ggtcgactcc acccaggagt tacgactgtt gccgaaagag agtttaccga
     5281 aactctgcga cgaactgcgc cgctatttac tcgacagcgt gagccgttcc agcgggcact
     5341 tcgcctccgg gctgggcacg gtcgaactga ccgtggcgct gcactatgtc tacaacaccc
     5401 cgtttgacca attgatttgg gatgtggggc atcaggctta tccgcataaa attttgaccg
     5461 gacgccgcga caaaatcggc accatccgtc agaaaggcgg tctgcacccg ttcccgtggc
     5521 gcggcgaaag cgaatatgac gtattaagcg tcgggcattc atcaacctcc attagtgccg
     5581 gaattggtat tgcggttgct gccgaaaaag aaggcaaaaa tcgccgcacc gtctgtgtca
     5641 ttggcgatgg cgcgattacc gcaggcatgg cgtttgaagc gatgaatcac gcgggcgata
     5701 tccgtcctga tatgctggtg attctcaacg acaatgaaat gtcgatttcc gaaaatgtcg
     5761 gcgcgctcaa caaccatctg gcacagctgc tttccggtaa gctttactct tcactgcgcg
     5821 aaggcgggaa aaaagttttc tctggcgtgc cgccaattaa agagctgctc aaacgcaccg
     5881 aagaacatat taaaggcatg gtagtgcctg gcacgttgtt tgaggagctg ggctttaact
     5941 acatcggccc ggtggacggt catgatgtgc tggggcttat caccacgcta aagaacatgc
     6001 gcgacctgaa aggcccgcag ttcctgcata tcatgaccaa aaaaggtcgt ggttatgaac
     6061 cggcagaaaa agacccgatc actttccatg ccgtgctaaa tttgatccct ccagcggttg
     6121 tttgccgaaa agtagcggcg gtttgccgag ctattcaaaa atctttggcg actggttgtg
     6181 cgaaacggca gcgaaagaca acaagctgat ggcgattact ccggcgatgc gtgaaggttc
     6241 cggcatggtc gagttttcac gtaaattccc ggatcgctac ttcgacgtgg caattgccga
     6301 gcaacatgcg gtgacctttg ctgcgggtct ggcgattggt gggtacaaac ccattgtcgc
     6361 gatttactcc actttcctgc aacgcgccta tgatcaggtg ctgcatgacg tggcgattca
     6421 aaagcttccg gtcctgttcg ccatcgaccg cgcgggcatt gttggtgctg acggtcaaac
     6481 ccatcagggt gcttttgatc tctcttacct gcgctgcata ccggaaatgg tcattatgac
     6541 cccgagcgat gaaaacgaat gtcgccagat gctctatacc ggctatcact ataacgatgg
     6601 cccgtcagcg gtgcgctacc cgcgtggcaa cgcggtcggc gtggaactga cgccgctgga
     6661 aaaactacca attggcaaag gcattgtgaa gcgtcgtggc gagaaactgg cgatccttaa
     6721 ctttggtacg ctgatgccag aagcggcgaa agtcgccgaa tcgctgaacg ccacgctggt
     6781 cgatatgcgt tttgtgaaac cgcttgatga agcgttaatt ctggaaatgg ccgccagcca
     6841 tgaagcgctg gtcaccgtag aagaaaacgc cattatgggc ggcgcaggca gcggcgtgaa
     6901 cgaagtgctg atggcccatc gtaaaccagt acccgtgctg aacattggcc tgccggactt
     6961 ctttattccg caaggaactc aggaagaaat gcgcgccgaa ctcggcctcg atgccgctgg
     7021 tatggaagcc aaaatcaagg cctggctggc ataagctttg tgtgaggaga tgtcgaatga
     7081 gcgcacatgc cctgcccaag gcagatctga ccgccaccag cctgatcgta tcgggcggca
     7141 tcatcgccgc ttggctggcc ctgcatgtgc atgcgctgtg gtttctggac gcagcggcgc
     7201 atcccatcct ggcgatcgca aatttcctgg ggctgacctg gctgtcggtc ggattgttca
     7261 tcatcgcgca tgacgcgatg cacgggtcgg tggtgccggg gcgtccgcgc gccaatgcgg
     7321 cgatgggcca gcttgtcctg tggctgtatg ccggattttc gtggcgcaag atgatcgtca
     7381 agcacatggc ccatcaccgc catgccggaa ccgacgacga ccccgatttc gaccatggcg
     7441 gcccggtccg ctggtacgcc cgcttcatcg gcacctattt cggctggcgc gaggggctgc
     7501 ttctgcccgt catcgtgacg gtctatgcgc tgatccttgg ggatcgctgg atgtacgtgg
     7561 tattctggcc gctgccgtcg atcctggcgt cgatccagct gttcgtgttc ggcacctggc
     7621 tgccgcaccg ccccggccac gacgcgttcc cggaccgcca caatgcgcgg tcgtcgcgga
     7681 tcagcgaccc cgtgtcgctg ctgacctgtt ttcactttgg cggttatcat cacgaacacc
     7741 acctgcaccc gacggtgccg tggtggcgcc tgcccagcac ccgcaccaag ggcgacaccg
     7801 cataagcttg agctgaggag attttgaatg caaacggaac acgtcatttt attgaatgca
     7861 cagggagttc ccacgggtac gctggaaaag tatgccgcac acacggcaga cacccgctta
     7921 catctcgcgt tctccagttg gctgtttaat gccaaaggac aattattagt tacccgccgc
     7981 gcactgagca aaaaagcatg gcctggcgtg tggactaact cggtttgtgg gcacccacaa
     8041 ctgggagaaa gcaacgagga cgcagtgatc cgccgttgcc gttatgagct tggcgtggaa
     8101 attacgcctc ctgaatctat ctatcctgac tttcgctacc gcgccaccga tccgagtggc
     8161 attgtggaaa atgaagtgtg tccggtattt gccgcacgca ccactagtgc gttacagatc
     8221 aatgatgatg aagtgatgga ttatcaatgg tgtgatttag cagatgtatt acacggtatt
     8281 gatgcaacgc cgtgggcgtt cagtccgtgg atggtgatgc aggcgacaaa tcgcgaagcc
     8341 agaaaacgat tatctgcatt tacccagctt aaataagctt tgcctcagga tttttgaaat
     8401 gacggtctgc gcaaaaaaac acgttcatct cactcgcgat gctgcggagc agttactggc
     8461 tgatattgat cgacgccttg atcagttatt gcccgtggag ggagaacggg atgttgtggg
     8521 tgccgcgatg cgtgaaggtg cgctggcacc gggaaaacgt attcgcccca tgttgctgtt
     8581 gctgaccgcc cgcgatctgg gttgcgctgt cagccatgac ggattactgg atttggcctg
     8641 tgcggtggaa atggtccacg cggcttcgct gatccttgac gatatgccct gcatggacga
     8701 tgcgaagctg cggcgcggac gccctaccat tcattctcat tacggagagc atgtggcaat
     8761 actggcggcg gttgccttgc tgagtaaagc ctttggcgta attgccgatg cagatggcct
     8821 cacgccgctg gcaaaaaatc gggcggtttc tgaactgtca aacgccatcg gcatgcaagg
     8881 attggttcag ggtcagttca aggatctgtc tgaaggggat aagccgcgca gcgctgaagc
     8941 tattttgatg acgaatcact ttaaaaccag cacgctgttt tgtgcctcga tgcagatggc
     9001 ctcgattgtt gcgaatgcct ccagcgaagc gcgtgattgc ctgcatcgtt tttcacttga
     9061 tcttggtcag gcatttcaac tgctggacga tttgaccgat ggcatgaccg acaccggtaa
     9121 ggatagcaat caggacgccg gtaaatcgac gctggtcaat ctgttaggcc cgagggcggt
     9181 tgaagaacgt ctgagacaac atcttcagct tgccagtgag catctctctg cggcctgcca
     9241 acacgggcac gccactcaac attttattca ggcctggttt gacaaaaaac tcgctgccgt
     9301 cagttaagct tgcaagaatt caagcttgga gtacgttaat tagttgtctg ttttaagccc
     9361 cgtgctctcc gccggggctt ttttattaaa aaccactcaa gtggtctcag cttgggatga
     9421 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     9481 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
     9541 ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     9601 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     9661 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     9721 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     9781 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     9841 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     9901 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     9961 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
    10021 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
    10081 acattcaaat atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat
    10141 gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa agccgtttct
    10201 gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt
    10261 ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa
    10321 ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt
    10381 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac
    10441 tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagtcga aatacgcgat
    10501 cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca
    10561 gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt
    10621 tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga
    10681 tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat
    10741 cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat
    10801 acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat
    10861 ataaatcagc atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa
    10921 tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg
    10981 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
    11041 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
    11101 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
    11161 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
    11221 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
    11281 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
    11341 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
    11401 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
    11461 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
    11521 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
    11581 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
    11641 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
    11701 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
    11761 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
    11821 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt
    11881 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
    11941 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
    12001 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgact ccgggagctg
    12061 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
    12121 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
    12181 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
    12241 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
    12301 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
    12361 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
    12421 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag
    12481 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt
    12541 ttccagactt tacgaaacac ggaaacccaa gaccattcat gttgttgctc aggtcgcaga
    12601 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
    12661 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
    12721 ccgtggggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
    12781 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
    12841 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
    12901 cacctgtcct acgagttgca tgataaagaa cacagtcata agtgcggcga cgatagtcat
    12961 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
    13021 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
    13081 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
    13141 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgacacg ggcaacagct
    13201 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
    13261 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtcct
    13321 cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa
    13381 tggcgcgcat tgcgcccagc gccatc
//
