LOCUS       sequence-190536-        8677 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ish1-mCherry insertion cassette with Hygro resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8677)
  AUTHORS   Gu M, LaJoie D, Chen OS, von Appen A, Ladinsky MS, Redd MJ, Nikolova
            L, Bjorkman PJ, Sundquist WI, Ullman KS, Frost A
  TITLE     LEM2 recruits CHMP7 for ESCRT-mediated nuclear envelope closure in 
            fission yeast and human cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Mar 14;114(11):E2166-E2175. doi: 
            10.1073/pnas.1613916114. Epub 2017 Feb 27.
  PUBMED    28242692
REFERENCE   2  (bases 1 to 8677)
  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"; doi: 
            "10.1073/pnas.1613916114"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2017-03-14- 14"; volume: "114"; issue: "11"; pages: 
            "E2166-E2175"
COMMENT     Sequence Label: pMGF157
FEATURES             Location/Qualifiers
     source          1..8677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(518..523,524..541,542..548)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    556..572
                     /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     580..596
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     580..596
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             592..1119
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(612..629)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(612..628)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(621..643)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1221..1226,1227..1242,1243..1249)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1257..1273
                     /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             1296..1949
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1907..1929
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1956..1979
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2574..3284
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2719..2737)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2987..3006
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      3305..3492
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3360..3384)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            3567..5142
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        3567..3910
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             3911..4939
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             3911..4939
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      4945..5142
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(5765..5787)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5932..5950)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6018..6122
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6123..6191,6192..6983)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6341..6360)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7154..7742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7643..7662
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7896..7913
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        7986..8063
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(8064..8677,1..469)
                     /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(8083..8102)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg
       61 ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc
      121 caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg
      181 tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc
      241 gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct
      301 gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa
      361 aagaaaaacc acgctagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
      421 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
      481 ttaatgtaag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
      541 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
      601 attacggatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
      661 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
      721 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg ctttgcctgg
      781 tttccggcac cagaagcggt gccggaaagc tggctggagt gcgatcttcc tgaggccgat
      841 actgtcgtcg tcccctcaaa ctggcagatg cacggttacg atgcgcccat ctacaccaac
      901 gtgacctatc ccattacggt caatccgccg tttgttccca cggagaatcc gacgggttgt
      961 tactcgctca catttaatgt tgatgaaagc tggctacagg aaggccagac gcgaattatt
     1021 tttgatggcg ttggaattag cttatcgact gcacggtgca ccaatgcttc tggcgtcagg
     1081 cagccatcgg aagctgtggt atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg
     1141 ctcaaggcgc actcccgttc tggataatgt tttttgcgcc gacatcataa cggttctggc
     1201 aaatattctg aaatgagctg ttgacaatta atcatcggct cgtataatgt gtggaattgt
     1261 gagcggataa caatttcaca caggaaacag tattcatgtc ccctatacta ggttattgga
     1321 aaattaaggg ccttgtgcaa cccactcgac ttcttttgga atatcttgaa gaaaaatatg
     1381 aagagcattt gtatgagcgc gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg
     1441 gtttggagtt tcccaatctt ccttattata ttgatggtga tgttaaatta acacagtcta
     1501 tggccatcat acgttatata gctgacaagc acaacatgtt gggtggttgt ccaaaagagc
     1561 gtgcagagat ttcaatgctt gaaggagcgg ttttggatat tagatacggt gtttcgagaa
     1621 ttgcatatag taaagacttt gaaactctca aagttgattt tcttagcaag ctacctgaaa
     1681 tgctgaaaat gttcgaagat cgtttatgtc ataaaacata tttaaatggt gatcatgtaa
     1741 cccatcctga cttcatgttg tatgacgctc ttgatgttgt tttatacatg gacccaatgt
     1801 gcctggatgc gttcccaaaa ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa
     1861 ttgataagta cttgaaatcc agcaagtata tagcatggcc tttgcagggc tggcaagcca
     1921 cgtttggtgg tggcgaccat cctccaaaat cggatctgga agttctgttc caggggcccc
     1981 tggctgttcc taattgggct gcttattttg atactttgag ctccaaagag ctttctgcct
     2041 ggattaagga atacaaaaaa cattaccatg gaaaacttca cccatccaaa gatcgtgaac
     2101 atcttttcca aaatgcctgt aatatttatc gcgctattgc tgaaaaggct gacaaatctg
     2161 ttcttcaatc gattcaatca aagtttccta aggcaactgt tcccggttat gactcttggt
     2221 ctaatgaaga tctaaagtct gcactcaagg aatatggtga cagtattggt aaggtgttta
     2281 atcgtaaaga tgctattgaa cgcttaaagc gtcatgatat tttattctat ggtcctgttg
     2341 ttgccgacaa ggctaaatca ggagtctctg gcttccttcg acgtgttgct tcttttatgg
     2401 gatttggcac tcgcgacgta gctgaaatta agctaggaga aaacattcaa aaggtaaaag
     2461 atactgcctc taaagcctca tctgctgtat cttcagcagg cgactatgtc gaaactaatg
     2521 taaagaaagc ccaaagggtt ttgggaggaa cgggcggtag tggaggcact ggtatggtga
     2581 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     2641 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     2701 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     2761 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     2821 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     2881 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     2941 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     3001 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     3061 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     3121 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     3181 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     3241 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtgaggcgcg ccacttctaa
     3301 ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa aaaaaataag
     3361 tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat tcttgagtaa
     3421 ctctttcctg taggtcaggt tgctttctca ggtatagtat gaggtcgctc ttattgacca
     3481 cacctctacc ggcagatccg ctagggataa cagggtaata tagatctgtt tagcttgcct
     3541 cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc cttgacagtc
     3601 ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag cccatacatc
     3661 cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga agcaaaaatt
     3721 acggctcctc gctgcggacc tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt
     3781 gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca ctgaggttct
     3841 tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc acatccgaac
     3901 ataaacaacc atgggtaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     3961 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     4021 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     4081 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
     4141 agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     4201 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     4261 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
     4321 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     4381 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     4441 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     4501 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     4561 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     4621 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     4681 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     4741 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     4801 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
     4861 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     4921 tccgagggca aaggaataat cagtactgac aataaaaaga ttcttgtttt caagaacttg
     4981 tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt tagcgtgatt
     5041 tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc gcagaaagta
     5101 atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa
     5161 cgccgccatc cataaattgt tcatgaacgt tcttaatgtc tatttatatt atttaaaaag
     5221 aaggagatta cgtctaaaac gagttctcgt ttaatatatg acttttaatg aaatatctat
     5281 gttcatattc gctacttaac cttccttggc aagtaatcca tcctattgtt aacattaata
     5341 taatccatca agatactttt ttattaattt aagtttttca gaattagatg ttggtgcacc
     5401 tcctccaaat ctagagctgt ggaaagaatt ggtaaaatct tatcgcgatg ttcgtgattc
     5461 tctaaaaatt gcagtaaaac atttctagca tattctttgt taatcaattc tttagaaata
     5521 ttgcttataa caggtggctc gaatgaacct gctgaatcac ttgaagcttt gggcaatatt
     5581 tcactgctca taacactgga cagctcgctc tcgtgagatt tgacactttc cttttgaacc
     5641 ttttgctttt caatgagttc ttttatttca tcccttgact ccactaactc gtttccaaca
     5701 ccgagcggcc gcatcgtgac tgactgacga tctgcctcgc gcgtttcggt gatgacggtg
     5761 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     5821 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
     5881 tgacccagtc acgtagcgat agcggagtgt ataattcttg aagacgaaag ggcctcgtga
     5941 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     6001 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     6061 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6121 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6181 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6241 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6301 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6361 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6421 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6481 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6541 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6601 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6661 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     6721 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6781 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6841 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6901 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6961 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     7021 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     7081 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     7141 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     7201 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     7261 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     7321 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     7381 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     7441 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7501 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7561 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7621 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7681 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     7741 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7801 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     7861 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     7921 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcataa
     7981 attccgacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
     8041 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
     8101 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
     8161 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
     8221 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
     8281 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
     8341 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
     8401 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
     8461 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
     8521 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
     8581 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
     8641 cggcgcgtct gcgtctggct ggctggcata aatatct
//
