LOCUS       pCDF-Frb-v1_addg       11331 bp DNA     circular SYN 02-MAY-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11331)
  AUTHORS   Richard Cooley
  TITLE     Direct Submission
  JOURNAL   Exported May 2, 2023 from SnapGene 6.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        55..68
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    69..74
                     /label=weak (4%)
     protein_bind    75..98
                     /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             113..135
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             129..134
                     /note="ribosome binding site"
     CDS             142..2805
                     /codon_start=1
                     /label=FrbA
                     /translation="MGRDLLRDLRLPDRNARYVSLAAAEEMGLVGISSAPRSLRVLVEN
                     TLRAAAAAKAGSTGDSLRQAVLSANSLIAAATANGPVSEVEFRPTRLVLQDHSGIPVLA
                     DLASLRSIMADRGLDPRLARPALPVDLVVDHSVEVASWARPDALSLNMRREYELNGERY
                     RFLRWAQQEISGLRVVPPGRGIVHQMHLEHLARVVLPDGDGLIAPDTVLGTDSHTPMVG
                     ALGVLGWGVGGIEAETVLLGHSVSLLSPQVIGVRLTGELPVGSTATDLALTMTEFLRNV
                     GVVGRFVEFFGPGVCALSVPDRATLSNMAPEYGCTVAFFPVDDTTLRYLELTGRGAEEI
                     RLVEAYCKEQGLFAGADASADPEFARVLDFDLSAVEPSAAGPRRPQDRLPLGTVPASFP
                     EPSGAALAGAAEGRRELADGAVAIAAITSCTNTANPHSMVTAGLVARKAVQRGLAVPSW
                     VKTSLAPGSRAVVHYLEAAGLLPDLEKLGFSVAGFGCTTCIGNSGGLAPEAEERARTEG
                     VRLAAVLSGNRNFEGRIHPDVAAAYLASPALVVAYALAGTVLTDLTARPVGTDPDGAEV
                     FLADLWPTPEEIDAAVGLVAREHFASEKESLFTGGPEWTSLEHPSGPVYDWPAESDYLL
                     RSPFFDGLTHAPLSDLSGARVLVLTGDSTTTDHISPAGAIGADSPAGRYLRALGVEPAL
                     FNSYGCRRGNHEVLVRGTFANPKFRNLLTPDVQGSSTLHVPTGERMSVHEAALAYAAEQ
                     VPVVVVGGRDYGFGSSRDWAAKGPALLGVRAVLAKSFERIHRSNLIGMGIVPLEFLPDQ
                     DAGTLGLTGHEALDVIGLDGLVPRGTVTVRARSAEGEPVAEWRMLARVDTAGELEYVRR
                     GGFLRSVASDLLDAEA"
     misc_feature    1501^1502
                     /label=1400bp
     misc_feature    2806..2811
                     /label=MCS
                     /note="multiple cloning site"
     CDS             2812..2829
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2896..2943
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2985..2998
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2999..3004
                     /label=moderate (10%)
     protein_bind    3005..3028
                     /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             3043..3065
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             3059..3064
                     /note="ribosome binding site"
     CDS             3072..4133
                     /codon_start=1
                     /label=frbB
                     /translation="MGTVSQYPDVAVRSDSPRPEASVLPAALELLESAGLTANWRIVAG
                     QAPDGDGGRKPEAGLLPPFAEPGASAYGLARAARRAWGGTFSVIERRGPLVRSDGTVNV
                     LLFRYELDMSDTGMEFESNSGEQLALVKTAAELGPREIPEQAALGIQAIAPDRVSEFVR
                     TAVGMCRRLDVDDLVVAHKGNSIKHTDGLFALTAAETLESLAGAAGIAWRSLIIDHMAA
                     ELVRNPLDYRAILTHSSFADLLAAALAGQMTTDQLGVCTTYDPAGRPLFGHDPALDTSA
                     PDADPERWRVRRLASTLAAGAALAEHLRLPPAGRALRRAAAELVRRDGGPIGWDQAVTT
                     ARKSLRAEGPARS"
     misc_feature    4135..4140
                     /label=MCS
                     /note="multiple cloning site"
     CDS             4141..4158
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4225..4272
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        4314..4327
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    4328..4333
                     /label=wt
     protein_bind    4334..4357
                     /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             4372..4394
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             4388..4393
                     /note="ribosome binding site"
     CDS             4401..5525
                     /codon_start=1
                     /label=frbC
                     /translation="MGRNDLVLEDTTLRDGEQTPGVAFSKETKTAILNALIEAGVTSIE
                     IGIPAMGGEELDFIKSVVDRQDEARLVVWHRGVREDVERSLDLGFTSVHVGLPTSAGHL
                     KASVRKDRTWLLATARDMVKMAKDRGAFVSISAEDIARTEISFLQEYAGVVAEAGADRL
                     RLSDTVGLLGPEAYGERVAAVLSAADIDVQCHAHNDFGLATANTLAGLKAGARYFHVTV
                     NAIGERAGMADLAQVVVALKKLYDRDLGIDLTKLKKVSRLVAEAAGHQVLPWQPITGDN
                     VFAHESGIHANGMFRDTSSFEPFPPEHVGGERRYVLGKHSGRALVAWALEQEGITPREE
                     LLPHCLEEVRALSIRIGGAVSHEQLVEIYNKAAA"
     misc_feature    5527..5532
                     /label=MCS
                     /note="multiple cloning site"
     CDS             5533..5550
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      5617..5664
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5706..5719
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    5720..5725
                     /label=wt
     protein_bind    5726..5749
                     /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             5764..5786
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             5780..5785
                     /note="ribosome binding site"
     CDS             5793..6626
                     /codon_start=1
                     /label=FrbD
                     /translation="MGTKRTMLRRAIARGGMSIAAGVHDGLSARIAQDAGFDVLWASGL
                     GISAAHAVPDDSILTMTEFLEAARVMNDSTDLPVLADCDTGFGDERNVARMVERYERAG
                     IAGVCIEDKVFPKRNSLGDGAQDQETIEGFAAKLAAAKKAQQTDDFVVVARIETFIAGG
                     VKEDAIARADAYVAAGADAILIHSKSRDGLEILDFGRSWQRKDVPLVAVPTTYPAVTAD
                     DLHEAGFSLAIYANQALRASITSMRESLRQITHDRSSLRIEPKVAKLSEVFELQR"
     misc_feature    6628..6633
                     /label=MCS
                     /note="multiple cloning site"
     CDS             6634..6651
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      6718..6765
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        6807..6820
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    6821..6826
                     /label=moderate (10%)
     protein_bind    6827..6850
                     /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             6865..6887
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             6881..6886
                     /note="ribosome binding site"
     CDS             6894..7994
                     /codon_start=1
                     /label=frbE
                     /translation="MGRKHTVTLIAGDGSGPELVAAARTVLDTAAGSAGFHLDWDVRPA
                     GQGALRSVGELLPASTLDSIRRNGVALKGPTTTPIGVGHRSVNITIRRELDLYACVRPV
                     RVWPGAGAVDSGIDVVVVRENTEDVYSGLEYPADSDGAAVVSATLSEFGHADPGIAFSL
                     KPITPGGSRRIVAFALDHARKNGRRKVEVGADVYRYPRTEGLFFEAARELGPEYPDIVT
                     ATVPVTVLCDSLVRDPADRDVLVLPNLYGDVISDVTAALVGGLGMAPGANIGDRTAVFE
                     ATHGSAPEFAGSNRLNPTALILSGCMLLDHLGEAPTAARIRAAVRSVLEEGRSVTFDLL
                     PPDRAARAVTTTAYAEAVIERLRDIG"
     misc_feature    7996..8001
                     /label=MCS
                     /note="multiple cloning site"
     CDS             8002..8019
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      8086..8133
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(8305..9096)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(9097..9188)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(9236..9974)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(10184..11266)
                     /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"
     promoter        complement(join(11267..11331,1..13))
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 attcgatggt gtccgggatc tcgacgctct ccctctggcg tctcaacacg aaattaatac
       61 gactcactat tattgaattg tgagcggata acaattcccc tctagaaata attttgttta
      121 actttaagaa ggagatatac catgggtcgt gatttactgc gcgatttacg cttaccggat
      181 cgtaatgcgc gttatgtgtc tttagcagct gccgaagaaa tggggttagt cggcatctca
      241 agcgcgccgc gtagcctccg tgtgctggtt gaaaacaccc tgagagccgc agcggctgca
      301 aaagcgggtt caaccggcga tagcctacgt caggccgtgc tgtctgcaaa tagcctgatc
      361 gctgcagcga cggcgaacgg cccggtgtct gaagttgagt tccgtccgac tcgcctggtc
      421 ctgcaagatc actcgggcat tccggtactt gccgacctgg cgtccctgcg tagtatcatg
      481 gctgatcgtg gtttggatcc gcgcctggca cgtcccgcac tcccggttga cttagtcgtt
      541 gatcattccg tggaagtggc cagctgggcg cgacctgacg ccttgtcctt aaacatgcgt
      601 cgcgaatatg aactgaacgg agagcggtat cgcttcctcc gttgggctca gcaggaaatc
      661 tctggactgc gtgtggttcc gccaggtcgt ggaattgtgc atcagatgca tcttgaacac
      721 ttggcgcgcg tggtgctgcc agacggtgat ggtctaatcg caccagatac ggtgttagga
      781 actgactctc acacaccgat ggtcggtgca ctaggcgttc tgggctgggg cgtaggtggc
      841 attgaagctg aaaccgtcct gctgggtcat agcgtttcct tactttcccc acaagtgatc
      901 ggcgttcgtt taaccggtga actgccggtg ggcagtacgg ctaccgactt ggcccttacc
      961 atgaccgagt ttcttcgcaa cgtgggagtt gtgggccggt ttgtggagtt tttcggccct
     1021 ggggtctgcg ccctgtctgt cccggacagg gccaccctaa gtaatatggc gccggaatac
     1081 ggttgcaccg tagcattctt cccggtcgac gatacgaccc tgcgctatct ggagctcacc
     1141 gggcgtgggg cagaagagat aaggcttgta gaggcttact gtaaagagca ggggctgttc
     1201 gcaggtgccg atgcgagcgc cgatccggaa tttgcccgag tgctggattt tgacctgtcc
     1261 gcggtcgagc cgtcggctgc ggggccgaga cggccccaag atcgtctgcc acttgggacc
     1321 gtaccggcgt cattccccga accgagcggt gccgcgttgg cgggcgcggc ggaagggcgg
     1381 cgtgaacttg cggacggcgc agttgcgatt gcggcaatta cctcatgcac caataccgcc
     1441 aatccgcaca gcatggtgac cgcggggttg gtcgcccgta aagccgtgca gcgcggcctc
     1501 gccgtgccga gttgggttaa gacgtcactt gcgccaggat cacgtgcggt cgtacactat
     1561 ttggaggcgg ccggcctgtt gccggatctg gaaaaactgg gattcagtgt ggcaggtttt
     1621 ggttgtacca cctgtattgg taacagcggc gggttagcac cggaagcgga agagcgcgct
     1681 cgtactgagg gagtccgcct tgctgccgtt ctaagtggca accgtaactt tgaaggtaga
     1741 atccatccgg atgtggctgc tgcgtacctg gccagtccgg ctttagtggt tgcgtatgcc
     1801 ttggctggca ccgttctgac cgatctaact gcccgtccgg ttggtacgga tcctgatggc
     1861 gcagaagtct ttctcgcgga cttatggccg acgccagagg agatcgatgc cgctgtgggt
     1921 ctggttgccc gcgagcactt cgcgtcagaa aaagaatcat tattcacggg aggtccggag
     1981 tggacgtcgc ttgagcatcc aagtgggcct gtatacgatt ggcccgcgga gagtgattat
     2041 ctgttacgtt ctcccttttt tgatggcctg acccatgcgc ctttatcgga tctttcgggg
     2101 gcccgtgttt tagtactgac tggtgatagc accacgactg accacatttc accagccggc
     2161 gctattggtg cagacagccc tgcaggccga tatctgcgtg cattaggcgt agaacccgct
     2221 ctttttaaca gttatggctg cagacggggc aaccatgaag tgttggtgcg cggcacattt
     2281 gcaaacccta aatttcgaaa cttgcttacc cctgatgttc agggctcatc aacgctgcat
     2341 gtgccgacag gtgaacgaat gagcgtccac gaagctgcgc tcgcttacgc agcagaacaa
     2401 gttcctgtag tggtggtagg gggccgcgat tacggtttcg gcagcagccg tgactgggcg
     2461 gctaaaggcc cggccttact gggtgtacgc gccgtgttag caaagtcctt tgaacgcatt
     2521 catcgctcga atttaatcgg tatgggcatt gtcccgctgg aatttctgcc cgatcaggac
     2581 gcgggcacgc tgggcttaac tggccatgag gctctcgatg tcattggact ggacggtcta
     2641 gttccgcgag gcacggtcac agttcgtgcc cgctccgcag agggcgagcc agtagctgaa
     2701 tggcggatgt tggcacgcgt agacaccgca ggcgaacttg aatacgtgcg tcgtggcggc
     2761 ttcctgcgtt ccgttgcgtc ggacctctta gacgccgaag cataactcga gcaccaccac
     2821 caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     2881 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     2941 ttgctgaaag gagttcgtga gacggagtcg tctcaacacg aaattaatac gactcactct
     3001 accagaattg tgagcggata acaattcccc tctagaaata attttgttta actttaagaa
     3061 ggagatatac catgggcacg gtgagccagt acccggatgt ggcagtgcgt tctgatagcc
     3121 cgcgcccgga agcctctgtg ctgccggccg cgctggaact gctggaatcg gccggattaa
     3181 cagctaactg gcgtattgtt gccggtcagg cgccggacgg cgatgggggt cgtaaacctg
     3241 aagcaggctt attaccgccg tttgccgaac cgggcgcgtc tgcgtatggt cttgcgcgcg
     3301 ccgcccgtcg tgcctggggc ggtactttca gcgttattga acgtcgcggc ccactggtcc
     3361 gctcggatgg caccgttaat gtcttgctct tccgctatga actggatatg agcgataccg
     3421 gcatggaatt tgaaagcaac tccggcgaac agctggcact ggttaaaacc gctgcggaac
     3481 tgggcccgcg tgaaattccg gaacaagcgg cgctaggcat tcaggcaata gcgccagatc
     3541 gtgtgtctga atttgtgcgc acggcggtgg gtatgtgccg ccggctcgat gttgatgatc
     3601 tggtggttgc gcacaaaggc aacagcatca aacataccga tggtctgttt gcgttaacgg
     3661 cagcggaaac gctggagtcc ctggcgggtg ccgcaggtat cgcatggcgc agcctgatta
     3721 ttgatcacat ggcggcggag ctggtgcgta acccgctgga ttatcgcgca attctgaccc
     3781 attcgagttt cgcagatctc ctagctgccg ctcttgccgg gcagatgacc acagatcaat
     3841 tgggtgtctg taccacctac gatcctgcgg gccgtccgtt atttggtcat gatccggcgc
     3901 ttgacaccag tgcaccggat gcagatcccg agcgctggcg tgttcgtcgt ctggcctcta
     3961 cgctcgcagc cggtgcagca cttgctgaac atctgcgtct cccgccagca ggtcgtgcgc
     4021 tgcgccgcgc ggcagcagag ttagttagaa gagacggtgg accaattgga tgggatcagg
     4081 ctgtaacgac cgcgaggaaa agtctacgtg ccgaaggccc tgcgcgtagc taaactcgag
     4141 caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg
     4201 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     4261 aggggttttt tgctgaaagg agttcgtgag acgcgctcgt ctcaacacga aattaatacg
     4321 actcactata ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa
     4381 ctttaagaag gagatatacc atgggccgta acgatctggt tctggaagat accacgctgc
     4441 gtgacggcga gcagaccccg ggcgtggcgt ttagcaaaga aaccaaaacc gcaatcctga
     4501 acgccctgat cgaagctggt gtcacctcaa ttgaaattgg catccccgcg atgggtggtg
     4561 aagagttaga ttttattaaa tctgtagtag accgccagga tgaagcccgc ctggtcgtct
     4621 ggcatcgcgg tgtgcgcgaa gacgttgaac gttcgctgga tcttggtttt acctctgttc
     4681 acgttggcct gccgaccagc gcgggccatc tgaaagcgag tgttcgtaaa gatcgtacct
     4741 ggctgctggc aacggcacgt gatatggtga aaatggcaaa agatagaggc gctttcgtta
     4801 gtatttcagc tgaagatatc gctcgcacag agatcagctt tctccaagaa tatgcggggg
     4861 ttgtggcaga agcgggtgct gatcgcttac gtttaagcga caccgtcggc ttgctcggac
     4921 cggaggctta tggcgagcgt gtggccgcag tattatccgc ggcggacatc gatgtgcagt
     4981 gtcatgcgca taacgatttc ggcctggcga ccgcaaacac tctggcgggt ttaaaggccg
     5041 gtgcgcgtta ttttcatgtt accgtaaatg cgattggaga acgtgccgga atggcggatc
     5101 tggcgcaggt tgtcgtagca ctaaaaaaac tttatgaccg cgatctcggc attgatttaa
     5161 cgaaactgaa aaaggtttcc cgacttgtcg ccgaagcggc cgggcatcag gtgctgccgt
     5221 ggcagccgat taccggcgat aacgtctttg ctcatgagtc cggtatccac gctaacggca
     5281 tgttccgcga cacgagcagt tttgaaccgt tcccgccgga acatgtgggc ggcgaacgcc
     5341 gctatgtcct tggcaaacat agcggtcgtg ctctggtggc ctgggcgttg gaacaggaag
     5401 ggattacgcc gcgtgaagaa ctgctcccgc attgcttaga agaagtgcgt gccctgtcga
     5461 ttcgtatcgg cggcgcggtt tcgcatgaac aactggtgga aatctacaac aaagccgcag
     5521 cgtaaactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag
     5581 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     5641 aaacgggtct tgaggggttt tttgctgaaa ggagttcgtg agacgtcatc gtctcaacac
     5701 gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat
     5761 aattttgttt aactttaaga aggagatata ccatgggcac caaacgcacc atgctgcgtc
     5821 gcgccatcgc gcgcggcggc atgagcattg ccgccggggt gcatgacggt ctgtccgccc
     5881 gtatcgccca ggatgcgggc tttgatgtgc tgtgggcgtc gggcctgggc atcagcgccg
     5941 ctcatgcggt gccggacgat tccattttga cgatgaccga atttctggag gcggctcgcg
     6001 taatgaatga ttctactgat cttccggtgc ttgcggattg cgataccggc ttcggcgatg
     6061 aacgtaatgt ggcgcgtatg gttgaacgct atgaacgggc gggcatcgca ggcgtctgca
     6121 ttgaggataa agtgtttccg aaacgtaact ccctgggtga tggtgcgcag gatcaggaaa
     6181 cgattgaagg ttttgcggcg aaattggcgg cagcaaaaaa agcgcaacaa accgatgact
     6241 tcgttgttgt cgcccgcatc gaaaccttta ttgccggagg tgtgaaagaa gatgccattg
     6301 cccgggccga cgcttacgtc gccgcaggtg ccgatgcaat tctgattcat agcaaaagcc
     6361 gcgatggcct ggaaatcctg gattttggtc gtagctggca gcgtaaagac gttccgctgg
     6421 tggccgttcc gaccacctat ccggcggtca ccgcggatga tctgcatgaa gcgggtttta
     6481 gcctggcgat ttacgccaac caggcgctgc gcgcgagtat tacgagcatg cgtgaaagcc
     6541 tgcgtcagat cacccatgat cgctcgtccc tgcgtattga gcccaaagtt gcgaaactga
     6601 gcgaagtgtt cgaactgcaa cgctaaactc gagcaccacc accaccacca ctgagatccg
     6661 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     6721 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggagttcgt
     6781 gagacgtgga cgtctcaaca cgaaattaat acgactcact ctaccagaat tgtgagcgga
     6841 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accatgggcc
     6901 gtaaacatac cgtgacgctg attgccggag atggcagcgg gccggaactg gtggcggcgg
     6961 ctcgtacggt actggatacc gcagccggtt cggcgggctt ccatttggac tgggatgtga
     7021 ggccagcagg tcagggcgct ctgcgtagcg ttggcgaact actcccggca tccaccctgg
     7081 acagcatccg gcgtaacggc gttgcactga aaggcccgac cacgactcca atcggcgtgg
     7141 gccatcgctc ggtgaatatt accatccgtc gcgagttaga tttgtacgcc tgtgtccgtc
     7201 cggtgcgtgt gtggccgggc gcgggtgcgg ttgattccgg cattgatgtt gtcgttgttc
     7261 gtgaaaacac cgaagatgtc tatagtggtc ttgagtatcc agctgatagc gacggcgcag
     7321 cggtggtgag tgccacctta agtgaatttg gtcatgcgga tccgggtatc gccttttcgt
     7381 tgaaaccgat tacaccgggc ggttcccgcc gtattgtcgc cttcgcgctg gatcatgcca
     7441 gaaaaaacgg tcgccgcaaa gtggaagtag gagcagatgt ttatcgctat ccgcgtaccg
     7501 aagggctttt ctttgaagca gcgcgtgagc tgggcccaga atacccggat atcgtgacgg
     7561 cgaccgtccc ggtgacagtc ttgtgtgatt ctctggtgcg cgacccagcg gatcgcgatg
     7621 tcttagtcct gccgaacctg tatggcgacg tcattagcga tgtgaccgcg gctctggtcg
     7681 gtggcctggg tatggcccct ggcgcgaaca ttggcgatcg taccgcggtg ttcgaagcga
     7741 cgcacggctc agcgccggag tttgccggta gcaaccgtct gaacccgacg gccctgatct
     7801 tgtccggctg catgctgttg gatcacctgg gtgaagcgcc gaccgccgct cgcattcgtg
     7861 cggcggtacg ctctgtgctg gaagaaggcc gcagcgtgac gtttgatctg ctgccgccgg
     7921 atcgtgccgc gcgcgcagtc accaccaccg catacgcgga ggcggtcatt gaaagattaa
     7981 gagacattgg ttaaactcga gcaccaccac caccaccact gagatccggc tgctaacaaa
     8041 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     8101 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gagttcgtga gacgatctca
     8161 cacggtcaca ctgcttccgg tagtcaataa accggtaaac cagcaataga cataagcggc
     8221 tatttaacga ccctgccctg aaccgacgac cgggtcatcg tggccggatc ttgcggcccc
     8281 tcggcttgaa cgaattgtta gacattattt gccgactacc ttggtgatct cgcctttcac
     8341 gtagtggaca aattcttcca actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc
     8401 aagataagcc tgtctagctt caagtatgac gggctgatac tgggccggca ggcgctccat
     8461 tgcccagtcg gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat
     8521 gcgggacaac gtaagcacta catttcgctc atcgccagcc cagtcgggcg gcgagttcca
     8581 tagcgttaag gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag
     8641 ttcctccgcc gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat
     8701 agccagatca atgtcgatcg tggctggctc gaagatacct gcaagaatgt cattgcgctg
     8761 ccattctcca aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg
     8821 cacaacaatg gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt
     8881 ttccaaaagg tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt
     8941 aaccagcaaa tcaatatcac tgtgtggctt caggccgcca tccactgcgg agccgtacaa
     9001 atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg acgccaacta cctctgatag
     9061 ttgagtcgat acttcggcga tcaccgcttc cctcatactc ttcctttttc aatattattg
     9121 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     9181 taaacaaata gctagctcac tcggtcgcta cgctccgggc gtgagactgc ggcgggcgct
     9241 gcggacacat acaaagttac ccacagattc cgtggataag caggggacta acatgtgagg
     9301 caaaacagca gggccgcgcc ggtggcgttt ttccataggc tccgccctcc tgccagagtt
     9361 cacataaaca gacgcttttc cggtgcatct gtgggagccg tgaggctcaa ccatgaatct
     9421 gacagtacgg gcgaaacccg acaggactta aagatcccca ccgtttccgg cgggtcgctc
     9481 cctcttgcgc tctcctgttc cgaccctgcc gtttaccgga tacctgttcc gcctttctcc
     9541 cttacgggaa gtgtggcgct ttctcatagc tcacacactg gtatctcggc tcggtgtagg
     9601 tcgttcgctc caagctgggc tgtaagcaag aactccccgt tcagcccgac tgctgcgcct
     9661 tatccggtaa ctgttcactt gagtccaacc cggaaaagca cggtaaaacg ccactggcag
     9721 cagccattgg taactgggag ttcgcagagg atttgtttag ctaaacacgc ggttgctctt
     9781 gaagtgtgcg ccaaagtccg gctacactgg aaggacagat ttggttgctg tgctctgcga
     9841 aagccagtta ccacggttaa gcagttcccc aactgactta accttcgatc aaaccacctc
     9901 cccaggtggt tttttcgttt acagggcaaa agattacgcg cagaaaaaaa ggatctcaag
     9961 aagatccttt gatcttttct actgaaccgc tctagatttc agtgcaattt atctcttcaa
    10021 atgtagcacc tgaagtcagc cccatacgat ataagttgta attctcatgt tagtcatgcc
    10081 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc
    10141 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca
    10201 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
    10261 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
    10321 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
    10381 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
    10441 tatcgtcgta tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg
    10501 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
    10561 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
    10621 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
    10681 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
    10741 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
    10801 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
    10861 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
    10921 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
    10981 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
    11041 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
    11101 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
    11161 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
    11221 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
    11281 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc c
//
