LOCUS       Exported               11836 bp ds-DNA     circular SYN 06-MAY-2024
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11836)
  AUTHORS   Jason Peters Lab
  TITLE     Direct Submission
  JOURNAL   Exported Monday, May 6, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11836
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     terminator      complement(3..51)
                     /label=pdhR-aceEF-lpd terminator
                     /note="color: #ffffff; direction: LEFT"
     terminator      complement(52..104)
                     /label=hisLGDCBHAFI terminator
                     /note="color: #ffffff; direction: LEFT"
     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."
                     /note="color: #ffffff"
     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)."
                     /note="color: #993366"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     terminator      1423..1476
                     /label=garPLRK-rnpB terminator
                     /note="color: #ffffff"
     terminator      1477..1533
                     /label=thrLABC terminator
                     /note="color: #ffffff"
     promoter        1580..1637
                     /label=synthetic promoter D
                     /note="color: #ffffff"
     misc_feature    1580..1601
                     /label=consensus UP
                     /note="from Estrem et al. 1998 PNAS; Identification of an
                     UP element consensus sequence forbacterial promoters"
                     /note="color: #a6acb3"
     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)."
                     /note="color: #31849b"
     misc_feature    1638..1680
                     /label=insert spacer here
                     /note="color: #a6acb3"
     ncRNA           1681..1773
                     /label=optimized sgRNA
                     /note="color: #8fbc8f; direction: RIGHT"
                     /ncRNA_class="other"
     terminator      1820..1870
                     /label=L3S1P13 terminator
                     /note="color: #ffffff"
     CDS             2029..2724
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /note="color: #993366"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     misc_feature    2096..2115
                     /label=mscar1 protospacer
                     /note="color: #a6acb3"
     misc_feature    2139..2158
                     /label=mscar2 protospacer
                     /note="color: #a6acb3"
     terminator      2779..2868
                     /label=spy terminator
                     /note="color: #ffffff"
     protein_bind    complement(2869..2916)
                     /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)."
                     /note="color: #31849b; direction: LEFT"
     promoter        2993..3021
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
                     /note="This feature has 3 segments:
                        1: 2993 .. 2998 / #ffffff
                        2: 2999 .. 3015 / #ffffff
                        3: 3016 .. 3021 / #ffffff"
     CDS             3132..3926
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=kanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /note="color: #993366"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    complement(3978..4025)
                     /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)."
                     /note="color: #31849b; direction: LEFT"
     terminator      4076..4162
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
                     /note="color: #ffffff"
     terminator      complement(4265..4359)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(4387..4403)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     mobile_element  4408..4605
                     /mobile_element_type="transposon:Tn7R"
                     /label=Tn7R
                     /note="color: #a6acb3"
     promoter        5083..5187
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             5188..6048
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 5188 .. 5256 / #ccffcc / signal sequence
                        2: 5257 .. 6048 / #ccffcc
                     Cleavage site after base 5256"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6349..6737
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
                     /note="color: #ffff00"
     oriT            7006..7114
                     /label=incP origin of transfer
                     /note="incP origin of transfer"
                     /note="color: #a6acb3"
     misc_feature    join(7218..11836,1..4605)
                     /label=Tn7 transposon
                     /note="color: #a6acb3"
     mobile_element  7218..7383
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
                     /note="color: #a6acb3"
     terminator      complement(7449..7503)
                     /label=L3S2P24 terminator
                     /note="L3S2P24 terminator"
                     /note="color: #ffffff; direction: LEFT"
     CDS             complement(7535..7634)
                     /codon_start=1
                     /label=3xMyc
                     /note="color: #993366"
                     /translation="GRGAETHL*RRSGTKVDFRRRSGTEAHL*GRSV"
     CDS             complement(7538..7627)
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /note="color: #cc99b2"
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             complement(7637..11740)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system. Novo CSE"
                     /label=dCas9-novo
                     /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"
                     /note="This feature has 5 segments:
                        1:  7637 ..  9220 / #993366
                        2:  9221 ..  9223 / #cc99b2
                        3:  9224 .. 11710 / #993366
                        4: 11711 .. 11713 / #cc99b2
                        5: 11714 .. 11740 / #993366"
                     /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"
     promoter        complement(11774..11831)
                     /label=Synthetic Promoter G
                     /note="color: #ffffff; direction: LEFT"
     protein_bind    11787..11803
                     /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)."
                     /note="color: #31849b"
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 ttgagaccaa ctttggtctc caccatagcg gtcggtctct
     1681 gtttaagagc tatgctggaa acagcatagc aagtttaaat aaggctagtc cgttatcaac
     1741 ttgaaaaagt ggcaccgagt cggtgctttt tttgaattca tgtggctgac cgttctgttg
     1801 tctctcgctc ttccgagtag acgaacaata aggcctccct aacggggggc cttttttatt
     1861 gataacaaaa gtcagtgctt ccgctatttc caaaataccg ggctaatacg gtttaaacga
     1921 aaatttatca aaaagagtat tgacttaaag tctaacctat aggatactta cagccacctc
     1981 ttaaagcttg ggaggcgagg taatactaca aaataaagga ggttttttat ggtgagcaag
     2041 ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc acatggaggg ctccatgaac
     2101 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     2161 gccaagctga aggtgaccaa gggtggcccc ctgcccttct cctgggacat cctgtcccct
     2221 cagttcatgt acggctccag ggccttcatc aagcaccccg ccgacatccc cgactactat
     2281 aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     2341 gccgtgaccg tgacccagga cacctccctg gaggacggca ccctgatcta caaggtgaag
     2401 ctccgcggca ccaacttccc tcctgacggc cccgtaatgc agaagaaaac aatgggctgg
     2461 gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc tgaagggcga cattaagatg
     2521 gccctgcgcc tgaaggacgg cggccgctac ctggcggact tcaagaccac ctacaaggcc
     2581 aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc gcaagttgga catcacctcc
     2641 cacaacgagg actacaccgt ggtggaacag tacgaacgct ccgagggccg ccactccacc
     2701 ggcggcatgg acgagctgta caagtaagac gtttaaacga cctcctggat ttgctcagac
     2761 agccttttcg tcattcgttt cagccaaaaa acttaagacc gccggtcttg tccactacct
     2821 tgcagtaatg cggtggacag gatcggcggt tttcttttct cttctcaaga agttcctata
     2881 ctttctagag aataggaact tcggaatagg aacttcctcc tgaacggcca taagaacgaa
     2941 ggctgtctgt tgaactctcg agccgtggaa acggatgaag gcacgaaccc agttgacata
     3001 agcctgttcg gttcgtaaac tgtaatgcaa gtagcgtatg cgctcaaaac tggatggctt
     3061 tcttgccgcc aaggatctga tggcgcaggg gatcaagatc tgatcaagag acaggatgag
     3121 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3181 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3241 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3301 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3361 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3421 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3481 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3541 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3601 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     3661 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3721 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3781 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3841 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     3901 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ctcgagcatc
     3961 agcttcaaaa gcgctctgaa gttcctatac tttctagaga ataggaactt cggaataggt
     4021 acttcaagat ccccaattcg agctcatcgt ccgggccgca agctcctagc ggcggatttg
     4081 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtctttcg
     4141 actgagcctt tcgttttatt tgatgcctca agctagagag tcattacccc aggcgtttaa
     4201 gggcaccaat aactgcctta aaaaaattac gccccgccct gccactcatc gcagtctagc
     4261 ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat
     4321 ggagttctga ggtcattact ggatctatca acaggagtcc aagctcagct aattaagcta
     4381 gcttatcgat accgtcgacc tcgaacccca cgcccctctt taatacgacg ggcaatttgc
     4441 acttcagaaa atgaagagtt tgctttagcc ataacaaaag tccagtatgc tttttcacag
     4501 cataactgga ctgatttcag tttacaacta ttctgtctag tttaagactt tattgtcata
     4561 gtttagatct attttgttca gtttaagact ttattgtccg cccacacctg cttacgcagg
     4621 gcatccattt attactcaac cgtaaccgat tttgccaggt tacgcggctg gtcctctagc
     4681 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     4741 ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4801 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     4861 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4921 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc gtcaccgaaa
     4981 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5041 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5101 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5161 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5221 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5281 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5341 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5401 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5461 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5521 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5581 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5641 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5701 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5761 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5821 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5881 aaatctggag ccggtgagcg tgggtcccgc ggtatcattg cagcactggg gccagatggt
     5941 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6001 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6061 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6121 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6181 tgagcgtcag accccgtaga aaagatcaaa ggcagttatt ggtgcctcac tgattaagca
     6241 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6301 ttaatttttg cggccgcaag atccggccac gatgcgtccg gcgtagagga tctgaagatc
     6361 agcagttcaa cctgttgata gtacgtacta agctctcatg tttcacgtac taagctctca
     6421 tgtttaacgt actaagctct catgtttaac gaactaaacc ctcatggcta acgtactaag
     6481 ctctcatggc taacgtacta agctctcatg tttcacgtac taagctctca tgtttgaaca
     6541 ataaaattaa tataaatcag caacttaaat agcctctaag gttttaagtt ttataagaaa
     6601 aaaaagaata tataaggctt ttaaagcttt taaggtttaa cggttgtgga caacaagcca
     6661 gggatgtaac gcactgagaa gcccttagag cctctcaaag caattttgag tgacacagga
     6721 acacttaacg gctgacatgg gaattagcca tggcatcaca gtatcgtgat gacagaggca
     6781 gggagtgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6841 cctgcgttat caggggattc cttaaggtat actttccgct gcataaccct gcttcggggt
     6901 cattatagcg attttttcgg tatatccatc ctttttcgca cgatatacag gattttgcca
     6961 aagggttcgt gtagactttc cttggtgtat ccaacggcgt cagccgggca ggataggtga
     7021 agtaggccca cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc acctggcggt
     7081 gctcaacggg aatcctgctc tgcgaggctg gccgataagc tctgataccg ctcgccgcag
     7141 ccgaacgacc gagcgcagcg ggtcagtgag cgaggaagcg gaagagccca aacataacag
     7201 gaagaaaaat gccccgctgt gggcggacaa aatagttggg aactgggagg ggtggaaatg
     7261 gagtttttaa ggattattta gggaagagtg acaaaataga tgggaactgg gtgtagcgtc
     7321 gtaagctaat acgaaaatta aaaatgacaa aatagtttgg aactagattt cacttatctg
     7381 gttcttgagg cggttaaaag agccgtactc ttctccgatg tcgactaggc catgatgctc
     7441 attctgtggg accaaaacga aaaaacaccc tttcgggtgt cttttctgga atttggtacc
     7501 gaggctgcaa gtcccattaa ggaggcgcgc cttattacag atcttcctca gagatgagct
     7561 tctgttccag atcttcttct gaaatcaact tttgttccag atcttcttca gagatgagtt
     7621 tctgctccgc ggccgcgtcg ccgcccagct gcgacaggtc gatgcgggtt tcgtacaggc
     7681 cggtgatcga ctgatggatc agggtggcgt ccaggacctc tttggtcgag gtgtagcgct
     7741 tgcggtcgat ggtggtgtcg aagtacttga aggcggccgg ggcgcccagg ttggtcaggg
     7801 tgaacaggtg gatgatgttc tcggcttgct cgcggatcgg cttgtcgcgg tgcttgttat
     7861 aggccgacag gaccttgtcc aggttggcgt cggccaggat gacgcgcttc gagaactccg
     7921 agatctgctc gatgatctcg tccaggtagt gcttgtgctg ctcgacgaac agctgcttct
     7981 gctcgttgtc ctccggcgag cccttcagct tctcgtagtg cgaggccagg tacaggaagt
     8041 tgacgtactt cgacggcagg gccagctcgt tgcccttctg cagctcgccg gcagaagcca
     8101 gcattcgctt ccggccgttt tctaactcga acagcgagta cttcggcagc ttgatgatca
     8161 ggtccttctt gacctctttg tagcccttgg cctccaggaa gtcgatcggg ttcttctcga
     8221 acgacgagcg ctccatgatg gtaatgccca gtaactcctt gaccgacttc agtttcttcg
     8281 acttgccctt ctcgaccttg gcgacgacca ggaccgagta ggcgacggtc ggcgagtcga
     8341 agccgccgta cttcttcggg tcccagtcct tcttgcgtgc gatcagcttg tccgagttgc
     8401 gctttggcag gatcgactcc ttcgagaagc cgccggtctg cacctcggtt ttcttgacaa
     8461 tgttgacttg cggcatgctt aagactttgc ggacggtggc gaagtcgcgg cccttgtccc
     8521 agacgatctc tccggtttcc ccgtttgtct cgatcagcgg gcgcttgcgg atctcgccgt
     8581 tggccagggt gatctcggtc ttgaagaagt tcatgatgtt cgagtagaag aagtacttgg
     8641 cggtggcctt gccgatctct tgctccgact tggcgatcat cttgcggacg tcgtagacct
     8701 tgtagtcgcc gtagacgaac tccgactcca gttttgggta cttcttgatc agggcggtgc
     8761 cgacgacggc gttcaggtag gcgtcgtggg cgtggtggta gttgttgatc tcgcggacct
     8821 tgtagaactg aaagtccttg cggaagtccg agacgagctt cgacttcagg gtgatgacct
     8881 tgacctcgcg aatcagctta tcgttctcgt cgtacttggt gttcatgcgc gagtccagga
     8941 tctgcgcgac gtgcttggtg atctgccggg tttcgaccag ctgccgcttg atgaagccgg
     9001 ccttgtccag ctccgacagg ccgccgcgct cggccttggt caggttgtcg aacttgcgct
     9061 gcgtgatcag cttggcgttc agcagctgcc gccagtagtt cttcatcttc ttaacgacct
     9121 cttccgacgg gacgttgtcc gacttgccgc ggttcttgtc cgagcgggtc aggaccttgt
     9181 tgtcgatgct atcgtccttc aggaaggact gcgggacgat ggcgtcgacg tcgtagtccg
     9241 acaggcggtt gatgtccagc tcttggtcga cgtacatgtc gcggccgttc tgcaggtagt
     9301 acaggtacag cttctcgttc tgcagctgcg tgttctcgac cgggtgctcc ttcaggatct
     9361 gcgagcccag ctccttgatg ccctcctcga tgcgcttcat gcgctcgcgc gaatttttct
     9421 gtcccttctg cgtggtctga ttctcgcggg ccatctcgat gacgatgttc tccggcttgt
     9481 ggcgtcccat taccttaacc agctcgtcga cgaccttgac ggtctgcagg atgcccttct
     9541 tgatggccgg cgagccggcc aggttggcga tgtgctcgtg cagcgagtcg ccttggcccg
     9601 agacttgggc cttctgaatg tcctccttga aggtcagcga gtcgtcgtgg atcagctgca
     9661 tgaagttgcg gttggcgaag ccgtccgact tcaggaagtc caggatggtc ttgcccgact
     9721 gcttgtcgcg gatgccgttg atcagcttgc gcgacaggcg gccccagccg gtgtagcggc
     9781 ggcgctttaa ctgcttcatg accttgtcgt cgaacaggtg ggcgtaggtc ttgaggcgct
     9841 cctcgatcat ctcgcggtcc tcgaacaggg tcagggtcag gacgatgtcc tccaggatgt
     9901 cctcgttctc ctcgttgtcc aggaagtcct tgtccttgat gatcttcagc aggtcgtggt
     9961 aggtgcccag cgaggcgttg aagcggtcct cgacgccgct gatctcgacc gagtcgaagc
    10021 actcgatctt cttgaagtag tcctctttca gctgcttgac ggtgaccttg cggttggtct
    10081 tgaacagcag gtcgacgatg gccttcttct gctcgcccga caggaaggcc ggcttgcgca
    10141 tgccctcggt gacgtacttg accttggtca gctcgttgta gacggtgaag tactcgtaca
    10201 gcaggctgtg cttaggcagg accttctcgt tcggcaggtt cttgtcgaag ttggtcatgc
    10261 gctcgatgaa cgactgcgcc gaggcgccct tgtcgacgac ctcttcgaag ttccacgggg
    10321 tgatggtttc ctccgacttg cgggtcatcc aggcgaagcg cgagttcccg cgggccagtg
    10381 gaccgacata gtacggtatg cgaaaggtca ggatcttctc gatcttctct cgattgtcct
    10441 tcaggaacgg gtagaagtcc tcttggcgtc tcaggatggc gtgcagctcg cccaggtgga
    10501 tctgatgcgg gatcgagccg ttgtcgaagg tgcgctgctt gcgcagcagg tcctcgcggt
    10561 tcagcttcac cagcagctcc tcggtgccgt ccatcttctc caggatcggc ttgatgaact
    10621 tgtagaactc ctcttgcgag gcgccgccgt cgatgtagcc ggcgtagccg ttcttcgact
    10681 gatcaaagaa gatctccttg tacttctccg gcagctgctg ccggaccagg gccttcagca
    10741 gggtcaggtc ttggtggtgc tcgtcgtagc gcttgatcat cgaggccgac agcggggcct
    10801 ttgtgatctc tgtgttgacg cgcaggatgt ctgacaggag gattgcgtcc gacaggttct
    10861 tggcggccag gaacaggtcg gcgtactgat cgccgatctg cgccagcagg ttgtccaggt
    10921 cgtcgtcgta ggtgtccttc gacagctgca gcttggcgtc ctcggccagg tcgaagttcg
    10981 acttgaagtt cggggtcagg cccagcgaca gggctatcaa gttgccgaac aggccgttct
    11041 tcttctcgcc cggcagttga gcaataagat tctccaggcg gcgcgacttc gacaggcggg
    11101 ccgacaggat ggccttagca tcaacgcccg aggcgttgat cgggttctcc tcgaacagct
    11161 gattgtaggt ctgcaccagc tgaatgaaca gcttgtcgac gtccgagttg tccgggttca
    11221 ggtcgccctc gatcaggaag tggccgcgga acttgatcat gtgggccagg gccaggtaga
    11281 tcaggcgcag gtcggccttg tcggtcgagt cgaccagctt cttgcgcagg tggtagatgg
    11341 tcggatactt ctcgtggtag gcgacctcgt cgacgatgtt gccgaagatc gggtggcgct
    11401 cgtgcttctt gtcctcctcg accaggaacg actcctccag gcggtggaag aacgagtcgt
    11461 cgaccttggc catctcgtta gaaaagatct cttgcaggta gcagatgcgg ttcttgcgcc
    11521 gcgtgtagcg gcgccgggcg gtcctcttca ggcgtgtggc ctcggccgtc tccccagaat
    11581 cgaatagcag cgccccgatc aaattcttct tgatcgagtg gcggtcggtg ttgcccagga
    11641 ccttgaactt cttcgacggg accttgtact cgtcggtgat gacggcccag ccgaccgagt
    11701 tggtgccgat ggccaggccg atcgagtact tcttgtccat actagtatag tcctccttct
    11761 cgctcttaat taaactagga ttataattgt tatccgctca caattcaagt gtttttcgaa
    11821 aaaaaatttc tactag
//
