LOCUS       sequence-437054-       11817 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MobileCRiSPRi test system encoded on a kanR Tn7 transposon; 
            mScarlet-I, PD-sgRNA (mscar targeting), and PC-dCas9-novo .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11817)
  AUTHORS   Hall AN, Hall BW, Kinney KJ, Olsen GG, Banta AB, Noguera DR, Donohue
            TJ, Peters JM
  TITLE     Tools for genetic engineering and gene expression control in 
            Novosphingobium aromaticivorans and Rhodobacter sphaeroides.
  JOURNAL   Appl Environ Microbiol. 2024 Sep 26:e0034824. doi: 
            10.1128/aem.00348-24.
  PUBMED    39324814
REFERENCE   2  (bases 1 to 11817)
  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: "Appl 
            Environ Microbiol. 2024 Sep 26:e0034824. doi: 10.1128/aem.00348-24."
COMMENT     Sequence Label: pJMP8823
FEATURES             Location/Qualifiers
     source          1..11817
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        171..248
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             249..1331
                     /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     complement(268..287)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    1608..1624
                     /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             2010..2705
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(2143..2161)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    complement(2850..2897)
                     /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)."
     promoter        join(2974..2979,2980..2996,2997..3002)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3113..3907
                     /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(3167..3186)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3777..3796
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    complement(3959..4006)
                     /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)."
     terminator      complement(4057..4143)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(4246..4340)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(4368..4384)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4369..4385)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4799..4818)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4918..4940
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4978..4996)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5064..5168
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5169..5237,5238..6029)
                     /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(5387..5406)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6330..6718
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            6987..7095
                     /note="incP origin of transfer"
     mobile_element  7199..7364
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     CDS             complement(7519..7608)
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             complement(join(7618..9201,9202..9204,9205..11691,
                     11692..11694,11695..11721))
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     protein_bind    11766..11785
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
ORIGIN
        1 tctgcagacg taaaaaaagc ggcgtggtta gccgcttttt taattgccgg agcgcaataa
       61 aaaagccccc ggaaggtgat cttccggggg ctttctcatg cgttgcttgt attgaaaatg
      121 ggggccatgc ctacctatct gcctcccggg aagcggcatg catttacgtt gacaccatcg
      181 aatggtgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg
      241 tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc
      301 agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag
      361 tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg
      421 gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc
      481 aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga
      541 tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac
      601 gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag
      661 ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca
      721 gtattatttt ctcccatgaa gatggtacgc gactgggcgt ggagcatctg gtcgcattgg
      781 gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc
      841 tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag
      901 gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg
      961 ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta
     1021 ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag
     1081 atagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc
     1141 aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc
     1201 tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct
     1261 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     1321 gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc accccacccg
     1381 ggtccttgag ccccattatc acctccaaag ccttctcgtc tggtcagttt cacctgtttt
     1441 acgtaaaaac ccgcttcggc gggtttttac ttttggggaa acacagaaaa aagcccgcac
     1501 ctgacagtgc gggctttttt tttcgaccaa aggtgcgact actcttgcct actacctatc
     1561 gactgagctg aaagaattcg gaaaattttt tttcaaaagt acttgaattg tgagcggata
     1621 acaattataa ttctagttag tctcgaactc gtggccgttc agtttaagag ctatgctgga
     1681 aacagcatag caagtttaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
     1741 tcggtgcttt ttttgaattc atgtggctga ccgttctgtt gtctctcgct cttccgagta
     1801 gacgaacaat aaggcctccc taacgggggg ccttttttat tgataacaaa agtcagtgct
     1861 tccgctattt ccaaaatacc gggctaatac ggtttaaacg aaaatttatc aaaaagagta
     1921 ttgacttaaa gtctaaccta taggatactt acagccacct cttaaagctt gggaggcgag
     1981 gtaatactac aaaataaagg aggtttttta tggtgagcaa gggcgaggca gtgatcaagg
     2041 agttcatgcg gttcaaggtg cacatggagg gctccatgaa cggccacgag ttcgagatcg
     2101 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     2161 agggtggccc cctgcccttc tcctgggaca tcctgtcccc tcagttcatg tacggctcca
     2221 gggccttcat caagcacccc gccgacatcc ccgactacta taagcagtcc ttccccgagg
     2281 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgccgtgacc gtgacccagg
     2341 acacctccct ggaggacggc accctgatct acaaggtgaa gctccgcggc accaacttcc
     2401 ctcctgacgg ccccgtaatg cagaagaaaa caatgggctg ggaagcgtcc accgagcggt
     2461 tgtaccccga ggacggcgtg ctgaagggcg acattaagat ggccctgcgc ctgaaggacg
     2521 gcggccgcta cctggcggac ttcaagacca cctacaaggc caagaagccc gtgcagatgc
     2581 ccggcgccta caacgtcgac cgcaagttgg acatcacctc ccacaacgag gactacaccg
     2641 tggtggaaca gtacgaacgc tccgagggcc gccactccac cggcggcatg gacgagctgt
     2701 acaagtaaga cgtttaaacg acctcctgga tttgctcaga cagccttttc gtcattcgtt
     2761 tcagccaaaa aacttaagac cgccggtctt gtccactacc ttgcagtaat gcggtggaca
     2821 ggatcggcgg ttttcttttc tcttctcaag aagttcctat actttctaga gaataggaac
     2881 ttcggaatag gaacttcctc ctgaacggcc ataagaacga aggctgtctg ttgaactctc
     2941 gagccgtgga aacggatgaa ggcacgaacc cagttgacat aagcctgttc ggttcgtaaa
     3001 ctgtaatgca agtagcgtat gcgctcaaaa ctggatggct ttcttgccgc caaggatctg
     3061 atggcgcagg ggatcaagat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     3121 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3181 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3241 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     3301 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     3361 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3421 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     3481 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     3541 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     3601 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     3661 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     3721 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     3781 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     3841 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     3901 cttctgagcg ggactctggg gttcgaaatg actcgagcat cagcttcaaa agcgctctga
     3961 agttcctata ctttctagag aataggaact tcggaatagg tacttcaaga tccccaattc
     4021 gagctcatcg tccgggccgc aagctcctag cggcggattt gtcctactca ggagagcgtt
     4081 caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat
     4141 ttgatgcctc aagctagaga gtcattaccc caggcgttta agggcaccaa taactgcctt
     4201 aaaaaaatta cgccccgccc tgccactcat cgcagtctag cttggattct caccaataaa
     4261 aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac
     4321 tggatctatc aacaggagtc caagctcagc taattaagct agcttatcga taccgtcgac
     4381 ctcgaacccc acgcccctct ttaatacgac gggcaatttg cacttcagaa aatgaagagt
     4441 ttgctttagc cataacaaaa gtccagtatg ctttttcaca gcataactgg actgatttca
     4501 gtttacaact attctgtcta gtttaagact ttattgtcat agtttagatc tattttgttc
     4561 agtttaagac tttattgtcc gcccacacct gcttacgcag ggcatccatt tattactcaa
     4621 ccgtaaccga ttttgccagg ttacgcggct ggtcctctag ctggcgtaat agcgaagagg
     4681 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
     4741 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta
     4801 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     4861 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4921 ggagctgcat gtgtcagagg ttttcaccgt cgtcaccgaa acgcgcgaga cgaaagggcc
     4981 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     5041 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5101 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5161 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5221 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5281 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5341 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5401 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5461 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5521 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5581 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5641 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5701 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5761 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5821 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5881 gtgggtcccg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5941 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6001 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     6061 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     6121 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6181 aaaagatcaa aggcagttat tggtgcctca ctgattaagc attggtaact gtcagaccaa
     6241 gtttactcat atatacttta gattgattta aaacttcatt tttaattttt gcggccgcaa
     6301 gatccggcca cgatgcgtcc ggcgtagagg atctgaagat cagcagttca acctgttgat
     6361 agtacgtact aagctctcat gtttcacgta ctaagctctc atgtttaacg tactaagctc
     6421 tcatgtttaa cgaactaaac cctcatggct aacgtactaa gctctcatgg ctaacgtact
     6481 aagctctcat gtttcacgta ctaagctctc atgtttgaac aataaaatta atataaatca
     6541 gcaacttaaa tagcctctaa ggttttaagt tttataagaa aaaaaagaat atataaggct
     6601 tttaaagctt ttaaggttta acggttgtgg acaacaagcc agggatgtaa cgcactgaga
     6661 agcccttaga gcctctcaaa gcaattttga gtgacacagg aacacttaac ggctgacatg
     6721 ggaattagcc atggcatcac agtatcgtga tgacagaggc agggagtgcg gcctttttac
     6781 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcaggggatt
     6841 ccttaaggta tactttccgc tgcataaccc tgcttcgggg tcattatagc gattttttcg
     6901 gtatatccat cctttttcgc acgatataca ggattttgcc aaagggttcg tgtagacttt
     6961 ccttggtgta tccaacggcg tcagccgggc aggataggtg aagtaggccc acccgcgagc
     7021 gggtgttcct tcttcactgt cccttattcg cacctggcgg tgctcaacgg gaatcctgct
     7081 ctgcgaggct ggccgataag ctctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7141 gggtcagtga gcgaggaagc ggaagagccc aaacataaca ggaagaaaaa tgccccgctg
     7201 tgggcggaca aaatagttgg gaactgggag gggtggaaat ggagttttta aggattattt
     7261 agggaagagt gacaaaatag atgggaactg ggtgtagcgt cgtaagctaa tacgaaaatt
     7321 aaaaatgaca aaatagtttg gaactagatt tcacttatct ggttcttgag gcggttaaaa
     7381 gagccgtact cttctccgat gtcgactagg ccatgatgct cattctgtgg gaccaaaacg
     7441 aaaaaacacc ctttcgggtg tcttttctgg aatttggtac cgaggctgca agtcccatta
     7501 aggaggcgcg ccttattaca gatcttcctc agagatgagc ttctgttcca gatcttcttc
     7561 tgaaatcaac ttttgttcca gatcttcttc agagatgagt ttctgctccg cggccgcgtc
     7621 gccgcccagc tgcgacaggt cgatgcgggt ttcgtacagg ccggtgatcg actgatggat
     7681 cagggtggcg tccaggacct ctttggtcga ggtgtagcgc ttgcggtcga tggtggtgtc
     7741 gaagtacttg aaggcggccg gggcgcccag gttggtcagg gtgaacaggt ggatgatgtt
     7801 ctcggcttgc tcgcggatcg gcttgtcgcg gtgcttgtta taggccgaca ggaccttgtc
     7861 caggttggcg tcggccagga tgacgcgctt cgagaactcc gagatctgct cgatgatctc
     7921 gtccaggtag tgcttgtgct gctcgacgaa cagctgcttc tgctcgttgt cctccggcga
     7981 gcccttcagc ttctcgtagt gcgaggccag gtacaggaag ttgacgtact tcgacggcag
     8041 ggccagctcg ttgcccttct gcagctcgcc ggcagaagcc agcattcgct tccggccgtt
     8101 ttctaactcg aacagcgagt acttcggcag cttgatgatc aggtccttct tgacctcttt
     8161 gtagcccttg gcctccagga agtcgatcgg gttcttctcg aacgacgagc gctccatgat
     8221 ggtaatgccc agtaactcct tgaccgactt cagtttcttc gacttgccct tctcgacctt
     8281 ggcgacgacc aggaccgagt aggcgacggt cggcgagtcg aagccgccgt acttcttcgg
     8341 gtcccagtcc ttcttgcgtg cgatcagctt gtccgagttg cgctttggca ggatcgactc
     8401 cttcgagaag ccgccggtct gcacctcggt tttcttgaca atgttgactt gcggcatgct
     8461 taagactttg cggacggtgg cgaagtcgcg gcccttgtcc cagacgatct ctccggtttc
     8521 cccgtttgtc tcgatcagcg ggcgcttgcg gatctcgccg ttggccaggg tgatctcggt
     8581 cttgaagaag ttcatgatgt tcgagtagaa gaagtacttg gcggtggcct tgccgatctc
     8641 ttgctccgac ttggcgatca tcttgcggac gtcgtagacc ttgtagtcgc cgtagacgaa
     8701 ctccgactcc agttttgggt acttcttgat cagggcggtg ccgacgacgg cgttcaggta
     8761 ggcgtcgtgg gcgtggtggt agttgttgat ctcgcggacc ttgtagaact gaaagtcctt
     8821 gcggaagtcc gagacgagct tcgacttcag ggtgatgacc ttgacctcgc gaatcagctt
     8881 atcgttctcg tcgtacttgg tgttcatgcg cgagtccagg atctgcgcga cgtgcttggt
     8941 gatctgccgg gtttcgacca gctgccgctt gatgaagccg gccttgtcca gctccgacag
     9001 gccgccgcgc tcggccttgg tcaggttgtc gaacttgcgc tgcgtgatca gcttggcgtt
     9061 cagcagctgc cgccagtagt tcttcatctt cttaacgacc tcttccgacg ggacgttgtc
     9121 cgacttgccg cggttcttgt ccgagcgggt caggaccttg ttgtcgatgc tatcgtcctt
     9181 caggaaggac tgcgggacga tggcgtcgac gtcgtagtcc gacaggcggt tgatgtccag
     9241 ctcttggtcg acgtacatgt cgcggccgtt ctgcaggtag tacaggtaca gcttctcgtt
     9301 ctgcagctgc gtgttctcga ccgggtgctc cttcaggatc tgcgagccca gctccttgat
     9361 gccctcctcg atgcgcttca tgcgctcgcg cgaatttttc tgtcccttct gcgtggtctg
     9421 attctcgcgg gccatctcga tgacgatgtt ctccggcttg tggcgtccca ttaccttaac
     9481 cagctcgtcg acgaccttga cggtctgcag gatgcccttc ttgatggccg gcgagccggc
     9541 caggttggcg atgtgctcgt gcagcgagtc gccttggccc gagacttggg ccttctgaat
     9601 gtcctccttg aaggtcagcg agtcgtcgtg gatcagctgc atgaagttgc ggttggcgaa
     9661 gccgtccgac ttcaggaagt ccaggatggt cttgcccgac tgcttgtcgc ggatgccgtt
     9721 gatcagcttg cgcgacaggc ggccccagcc ggtgtagcgg cggcgcttta actgcttcat
     9781 gaccttgtcg tcgaacaggt gggcgtaggt cttgaggcgc tcctcgatca tctcgcggtc
     9841 ctcgaacagg gtcagggtca ggacgatgtc ctccaggatg tcctcgttct cctcgttgtc
     9901 caggaagtcc ttgtccttga tgatcttcag caggtcgtgg taggtgccca gcgaggcgtt
     9961 gaagcggtcc tcgacgccgc tgatctcgac cgagtcgaag cactcgatct tcttgaagta
    10021 gtcctctttc agctgcttga cggtgacctt gcggttggtc ttgaacagca ggtcgacgat
    10081 ggccttcttc tgctcgcccg acaggaaggc cggcttgcgc atgccctcgg tgacgtactt
    10141 gaccttggtc agctcgttgt agacggtgaa gtactcgtac agcaggctgt gcttaggcag
    10201 gaccttctcg ttcggcaggt tcttgtcgaa gttggtcatg cgctcgatga acgactgcgc
    10261 cgaggcgccc ttgtcgacga cctcttcgaa gttccacggg gtgatggttt cctccgactt
    10321 gcgggtcatc caggcgaagc gcgagttccc gcgggccagt ggaccgacat agtacggtat
    10381 gcgaaaggtc aggatcttct cgatcttctc tcgattgtcc ttcaggaacg ggtagaagtc
    10441 ctcttggcgt ctcaggatgg cgtgcagctc gcccaggtgg atctgatgcg ggatcgagcc
    10501 gttgtcgaag gtgcgctgct tgcgcagcag gtcctcgcgg ttcagcttca ccagcagctc
    10561 ctcggtgccg tccatcttct ccaggatcgg cttgatgaac ttgtagaact cctcttgcga
    10621 ggcgccgccg tcgatgtagc cggcgtagcc gttcttcgac tgatcaaaga agatctcctt
    10681 gtacttctcc ggcagctgct gccggaccag ggccttcagc agggtcaggt cttggtggtg
    10741 ctcgtcgtag cgcttgatca tcgaggccga cagcggggcc tttgtgatct ctgtgttgac
    10801 gcgcaggatg tctgacagga ggattgcgtc cgacaggttc ttggcggcca ggaacaggtc
    10861 ggcgtactga tcgccgatct gcgccagcag gttgtccagg tcgtcgtcgt aggtgtcctt
    10921 cgacagctgc agcttggcgt cctcggccag gtcgaagttc gacttgaagt tcggggtcag
    10981 gcccagcgac agggctatca agttgccgaa caggccgttc ttcttctcgc ccggcagttg
    11041 agcaataaga ttctccaggc ggcgcgactt cgacaggcgg gccgacagga tggccttagc
    11101 atcaacgccc gaggcgttga tcgggttctc ctcgaacagc tgattgtagg tctgcaccag
    11161 ctgaatgaac agcttgtcga cgtccgagtt gtccgggttc aggtcgccct cgatcaggaa
    11221 gtggccgcgg aacttgatca tgtgggccag ggccaggtag atcaggcgca ggtcggcctt
    11281 gtcggtcgag tcgaccagct tcttgcgcag gtggtagatg gtcggatact tctcgtggta
    11341 ggcgacctcg tcgacgatgt tgccgaagat cgggtggcgc tcgtgcttct tgtcctcctc
    11401 gaccaggaac gactcctcca ggcggtggaa gaacgagtcg tcgaccttgg ccatctcgtt
    11461 agaaaagatc tcttgcaggt agcagatgcg gttcttgcgc cgcgtgtagc ggcgccgggc
    11521 ggtcctcttc aggcgtgtgg cctcggccgt ctccccagaa tcgaatagca gcgccccgat
    11581 caaattcttc ttgatcgagt ggcggtcggt gttgcccagg accttgaact tcttcgacgg
    11641 gaccttgtac tcgtcggtga tgacggccca gccgaccgag ttggtgccga tggccaggcc
    11701 gatcgagtac ttcttgtcca tactagtata gtcctccttc tcgctcttaa ttaaactaga
    11761 attataattg tgagcgctca caatttcaag tacttttgaa aaaaaatttt ccactag
//
