LOCUS       sequence-309441-       12490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid for insertion of the lacIZ operon and lac promoter 
            into the S. Typhimurium chromosome.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12490)
  AUTHORS   Holden ER, Wickham GJ, Webber MA, Thomson NM, Trampari E
  TITLE     Donor plasmids for phenotypically neutral chromosomal gene 
            insertions in Enterobacteriaceae.
  JOURNAL   Microbiology (Reading). 2020 Nov 23. doi: 10.1099/mic.0.000994.
  PUBMED    33226934
REFERENCE   2  (bases 1 to 12490)
  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: 
            "Microbiology (Reading). 2020 Nov 23. doi: 10.1099/mic.0.000994."
COMMENT     Sequence Label: pDOC-K-glmS-lacIZ
FEATURES             Location/Qualifiers
     source          1..12490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1819
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1720..1739
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1973..1990
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2107..2128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2143..2148,2149..2166,2167..2173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2181..2197
                     /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)."
     primer_bind     2205..2221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2724..2771)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             2981..3775
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3035..3054)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3645..3664
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    complement(3966..3999)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(4059..7133)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYG
                     QDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQ
                     ISDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIID
                     ERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIEN
                     GLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLW
                     YTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWS
                     LGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVP
                     KWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSL
                     IKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQ
                     TIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQ
                     LWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELG
                     NKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWK
                     AAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDV
                     EVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPY
                     VFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDG
                     FHMGIGGDDSWSPSVSAEFQLSAGRYHYQLVWCQK"
     primer_bind     7097..7113
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7129..7145)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7153..7169
                     /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)."
     promoter        complement(join(7177..7183,7184..7201,7202..7207))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7222..7243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(7256..8338)
                     /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     8300..8319
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(8914..8930)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9110..9127)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      9280..9366
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      9458..9485
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     oriT            9932..10041
                     /note="incP origin of transfer"
     promoter        10415..10860
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(10861..10947,10948..12282)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     primer_bind     complement(12321..12340)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12440..12462
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta gccaccactt caagaactct gtagcaccgc ctacatacct
     1441 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1501 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1561 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1621 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1681 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     1741 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     1801 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     1861 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     1921 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     1981 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     2041 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     2101 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     2161 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     2221 ccatgattac gccaagctct agggataaca gggtaatcga tgaattcgct cgaaggcgcg
     2281 ctgaagctga aagagatctc ttacattcac gcggaagcct atgctgcagg cgagctgaaa
     2341 cacgggccgt tggcgctgat tgacgccgat atgccggtga ttgtggtggc gccgaacaac
     2401 gaactgctgg agaaactgaa atccaacatc gaagaagttc gcgcacgtgg cggtcagttg
     2461 tacgtcttcg ccgatcagga tgcgggtttc gtcagcaacg acaacatgca catcatcgag
     2521 atgccgcatg tggaagaggt cattgcgcct atcttctaca ccgttccgct gcaactgctg
     2581 gcgtatcacg tggcgctgat taaaggcacc gacgttgacc agccgcgtaa cctggcgaaa
     2641 tcggttaccg tagagtaagt acaaatgtag taccaggcag aaggccgggt acctaggacc
     2701 ggtcaattgg ctggagctgc ttcgaagttc ctatactttc tagagaatag gaacttcgga
     2761 ataggaactt caagatcccc cacgctgccg caagcactca gggcgcaagg gctgctaaag
     2821 gaagcggaac acgtagaact taagggaatt gccagctggg gcgccctctg gtaaggttgg
     2881 gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg
     2941 atcaagatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3001 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3061 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3121 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3181 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3241 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3301 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3361 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3421 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3481 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     3541 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3601 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3661 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgctct acggtatcgc
     3721 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     3781 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     3841 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     3901 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccag cttcaaaagc
     3961 gctctgaagt tcctatactt tctagagaat aggaacttcg gaataggaac taaggaggat
     4021 attcatatct cgagcatatg ccccccaagc ttctcgagtt atttttgaca ccagaccaac
     4081 tggtaatggt agcgaccggc gctcagctgg aattccgccg atactgacgg gctccaggag
     4141 tcgtcgccac caatccccat atggaaaccg tcgatattca gccatgtgcc ttcttccgcg
     4201 tgcagcagat ggcgatggct ggtttccatc agttgctgtt gactgtagcg gctgatgttg
     4261 aactggaagt cgccgcgcca ctggtgtggg ccataattca attcgcgcgt cccgcagcgc
     4321 agaccgtttt cgctcgggaa gacgtacggg gtatacatgt ctgacaatgg cagatcccag
     4381 cggtcaaaac aggcggcagt aaggcggtcg ggatagtttt cttgcggccc taatccgagc
     4441 cagtttaccc gctctgctac ctgcgccagc tggcagttca ggccaatccg cgccggatgc
     4501 ggtgtatcgc tcgccacttc aacatcaacg gtaatcgcca tttgaccact accatcaatc
     4561 cggtaggttt tccggctgat aaataaggtt ttcccctgat gctgccacgc gtgagcggtc
     4621 gtaatcagca ccgcatcagc aagtgtatct gccgtgcact gcaacaacgc tgcttcggcc
     4681 tggtaatggc ccgccgcctt ccagcgttcg acccaggcgt tagggtcaat gcgggtcgct
     4741 tcacttacgc caatgtcgtt atccagcggt gcacgggtga actgatcgcg cagcggcgtc
     4801 agcagttgtt ttttatcgcc aatccacatc tgtgaaagaa agcctgactg gcggttaaat
     4861 tgccaacgct tattacccag ctcgatgcaa aaatccattt cgctggtggt cagatgcggg
     4921 atggcgtggg acgcggcggg gagcgtcaca ctgaggtttt ccgccagacg ccactgctgc
     4981 caggcgctga tgtgcccggc ttctgaccat gcggtcgcgt tcggttgcac tacgcgtact
     5041 gtgagccaga gttgcccggc gctctccggc tgcggtagtt caggcagttc aatcaactgt
     5101 ttaccttgtg gagcgacatc cagaggcact tcaccgcttg ccagcggctt accatccagc
     5161 gccaccatcc agtgcaggag ctcgttatcg ctatgacgga acaggtattc gctggtcact
     5221 tcgatggttt gcccggataa acggaactgg aaaaactgct gctggtgttt tgcttccgtc
     5281 agcgctggat gcggcgtgcg gtcggcaaag accagaccgt tcatacagaa ctggcgatcg
     5341 ttcggcgtat cgccaaaatc accgccgtaa gccgaccacg ggttgccgtt ttcatcatat
     5401 ttaatcagcg actgatccac ccagtcccag acgaagccgc cctgtaaacg gggatactga
     5461 cgaaacgcct gccagtattt agcgaaaccg ccaagactgt tacccatcgc gtgggcgtat
     5521 tcgcaaagga tcagcgggcg cgtctctcca ggtagcgaaa gccatttttt gatggaccat
     5581 ttcggcacag ccgggaaggg ctggtcttca tccacgcgcg cgtacatcgg gcaaataata
     5641 tcggtggccg tggtgtcggc tccgccgcct tcatactgca ccgggcggga aggatcgaca
     5701 gatttgatcc agcgatacag cgcgtcgtga ttagcgccgt ggcctgattc attccccagc
     5761 gaccagatga tcacactcgg gtgattacga tcgcgctgca ccattcgcgt tacgcgttcg
     5821 ctcatcgccg gtagccagcg cggatcatcg gtcagacgat tcattggcac catgccgtgg
     5881 gtttcaatat tggcttcatc caccacatac aggccgtagc ggtcgcacag cgtgtaccac
     5941 agcggatggt tcggataatg cgaacagcgc acggcgttaa agttgttctg cttcatcagc
     6001 aggatatcct gcaccatcgt ctgctcatcc atgacctgac catgcagagg atgatgctcg
     6061 tgacggttaa cgcctcgaat cagcaacggc ttgccgttca gcagcagcag accattttca
     6121 atccgcacct cgcggaaacc gacatcgcag gcttctgctt caatcagcgt gccgtcggcg
     6181 gtgtgcagtt caaccaccgc acgatagaga ttcgggattt cggcgctcca cagtttcggg
     6241 ttttcgacgt tcagacgtag tgtgacgcga tcggcataac caccacgctc atcgataatt
     6301 tcaccgccga aaggcgcggt gccgctggcg acctgcgttt caccctgcca taaagaaact
     6361 gttacccgta ggtagtcacg caactcgccg cacatctgaa cttcagcctc cagtacagcg
     6421 cggctgaaat catcattaaa gcgagtggca acatggaaat cgctgatttg tgtagtcggt
     6481 ttatgcagca acgagacgtc acggaaaatg ccgctcatcc gccacatatc ctgatcttcc
     6541 agataactgc cgtcactcca gcgcagcacc atcaccgcga ggcggttttc tccggcgcgt
     6601 aaaaatgcgc tcaggtcaaa ttcagacggc aaacgactgt cctggccgta accgacccag
     6661 cgcccgttgc accacagatg aaacgccgag ttaacgccat caaaaataat tcgcgtctgg
     6721 ccttcctgta gccagctttc atcaacatta aatgtgagcg agtaacaacc cgtcggattc
     6781 tccgtgggaa caaacggcgg attgaccgta atgggatagg tcacgttggt gtagatgggc
     6841 gcatcgtaac cgtgcatctg ccagtttgag gggacgacga cagtatcggc ctcaggaaga
     6901 tcgcactcca gccagctttc cggcaccgct tctggtgccg gaaaccaggc aaagcgccat
     6961 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     7021 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     7081 tcccagtcac gacgttgtaa aacgacggcc agtgaatccg taatcatggt catagctgtt
     7141 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     7201 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     7261 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     7321 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     7381 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     7441 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     7501 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg
     7561 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     7621 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     7681 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     7741 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     7801 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     7861 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     7921 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     7981 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     8041 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     8101 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     8161 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     8221 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     8281 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     8341 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     8401 gccattcgat ggtgcggccg ctattgaggg atctggggcg gccgcaagct ttcgacagac
     8461 ggcctttttt tgtgcgctgt gacaggcgct gttctttctc tatggtggct gaagagcgac
     8521 taagaggtgc tacgcctgcc tgcgctacgg gcacccggtc acattctctt ctttaaggcc
     8581 ctggctcaat ccaggtggat cggaaatcaa accaccccag gttattcagg tggatccccc
     8641 tggcccggtc ggggcttttg agtcgcatcc agtggtgaaa aagcggctgt agccagcctg
     8701 aacgggtggt ttccctgacc agttgttccg cgctgatggt ttcggcatgc cactgggttt
     8761 cccattgggc cagcagcgca tcatccccac cactgatggc gtggcgcagt aaaggggagc
     8821 ccagtagcca tgtaccgacg ttccactctt cgggaatggg gaaattgacc ggctagcgtc
     8881 gactagtagg gataacaggg taatgagctt ggcactggcc gtcgttttac aacgtcgtga
     8941 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     9001 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     9061 tggcgaatgg cgagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga
     9121 ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg
     9181 tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg
     9241 tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
     9301 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     9361 acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca
     9421 ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc
     9481 ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa atatgcatgc
     9541 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatcgacat
     9601 ccgccctcac cgccaggaac gcaaccgcag cctcatcacg ccggcgcttc ttggccgcgc
     9661 gggattcaac ccactcggcc agctcgtcgg tgtagctctt tggcatcgtc tctcgcctgt
     9721 cccctcagtt cagtaatttc ctgcatttgc ctgtttccag tcggtagata ttccacaaaa
     9781 cagcagggaa gcagcgcttt tccgctgcat aaccctgctt cggggtcatt atagcgattt
     9841 tttcggtata tccatccttt ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag
     9901 actttccttg gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta ggcccacccg
     9961 cgagcgggtg ttccttcttc actgtccctt attcgcacct ggcggtgctc aacgggaatc
    10021 ctgctctgcg aggctggccg gctaccgccg gcgtaacaga tgagggcaag cggatggctg
    10081 atgaaaccaa gccaaccagg aagggcagcc cacctatcaa ggtgtactgc cttccagacg
    10141 aacgaagagc gattgaggaa aaggcggcgg cggccggcat gagcctgtcg gcctacctgc
    10201 tggccgtcgg ccagggctac aaaatcacgg gcgtcgtgga ctatgagcac gtccgcgagc
    10261 tggcccgcat caatggcgac ctgggccgcc tgggcggcct gctgaaactc tggctcaccg
    10321 acgacccgcg cacggcgcgg ttcggtgatg ccacgatcct cgccctgctg gcgaagatcg
    10381 actctagcta gaggatcgat cctttttaac ccatcacata tacctgccgt tcactattat
    10441 ttagtgaaat gagatattat gatattttct gaattgtgat taaaaaggca actttatgcc
    10501 catgcaacag aaactataaa aaatacagag aatgaaaaga aacagataga ttttttagtt
    10561 ctttaggccc gtagtctgca aatcctttta tgattttcta tcaaacaaaa gaggaaaata
    10621 gaccagttgc aatccaaacg agagtctaat agaatgaggt cgaaaagtaa atcgcgcggg
    10681 tttgttactg ataaagcagg caagacctaa aatgtgtaaa gggcaaagtg tatactttgg
    10741 cgtcacccct tacatatttt aggtcttttt ttattgtgcg taactaactt gccatcttca
    10801 aacaggaggg ctggaagaag cagaccgcta acacagtaca taaaaaagga gacatgaacg
    10861 atgaacatca aaaagtttgc aaaacaagca acagtattaa cctttactac cgcactgctg
    10921 gcaggaggcg caactcaagc gtttgcgaaa gaaacgaacc aaaagccata taaggaaaca
    10981 tacggcattt cccatattac acgccatgat atgctgcaaa tccctgaaca gcaaaaaaat
    11041 gaaaaatatc aagttcctga gttcgattcg tccacaatta aaaatatctc ttctgcaaaa
    11101 ggcctggacg tttgggacag ctggccatta caaaacgctg acggcactgt cgcaaactat
    11161 cacggctacc acatcgtctt tgcattagcc ggagatccta aaaatgcgga tgacacatcg
    11221 atttacatgt tctatcaaaa agtcggcgaa acttctattg acagctggaa aaacgctggc
    11281 cgcgtcttta aagacagcga caaattcgat gcaaatgatt ctatcctaaa agaccaaaca
    11341 caagaatggt caggttcagc cacatttaca tctgacggaa aaatccgttt attctacact
    11401 gatttctccg gtaaacatta cggcaaacaa acactgacaa ctgcacaagt taacgtatca
    11461 gcatcagaca gctctttgaa catcaacggt gtagaggatt ataaatcaat ctttgacggt
    11521 gacggaaaaa cgtatcaaaa tgtacagcag ttcatcgatg aaggcaacta cagctcaggc
    11581 gacaaccata cgctgagaga tcctcactac gtagaagata aaggccacaa atacttagta
    11641 tttgaagcaa acactggaac tgaagatggc taccaaggcg aagaatcttt atttaacaaa
    11701 gcatactatg gcaaaagcac atcattcttc cgtcaagaaa gtcaaaaact tctgcaaagc
    11761 gataaaaaac gcacggctga gttagcaaac ggcgctctcg gtatgattga gctaaacgat
    11821 gattacacac tgaaaaaagt gatgaaaccg ctgattgcat ctaacacagt aacagatgaa
    11881 attgaacgcg cgaacgtctt taaaatgaac ggcaaatggt atctgttcac tgactcccgc
    11941 ggatcaaaaa tgacgattga cggcattacg tctaacgata tttacatgct tggttatgtt
    12001 tctaattctt taactggccc atacaagccg ctgaacaaaa ctggccttgt gttaaaaatg
    12061 gatcttgatc ctaacgatgt aacctttact tactcacact tcgctgtacc tcaagcgaaa
    12121 ggaaacaatg tcgtgattac aagctatatg acaaacagag gattctacgc agacaaacaa
    12181 tcaacgtttg cgcctagctt cctgctgaac atcaaaggca agaaaacatc tgttgtcaaa
    12241 gacagcatcc ttgaacaagg acaattaaca gttaacaaat aaaaacgcaa aagaaaatgc
    12301 cgattatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
    12361 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
    12421 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
    12481 aaacgcgcga
//
