LOCUS       Exported                6994 bp ds-DNA     circular SYN 07-MAY-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6994)
  AUTHORS   alexbrown
  TITLE     Direct Submission
  JOURNAL   Exported Monday, May 11, 2020 from SnapGene Viewer 5.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6994
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        3..80
                     /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             81..1163
                     /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(100..119)
                     /label=MK 6810
                     /note="[-] LacI::LacI-R"
     primer_bind     1100..1116
                     /label=MK 1197
                     /note="[-] OS-A:NotI::OSdup-del Sequencing Primer-at 
                     Approx. 25"
     primer_bind     1180..1203
                     /label=MK 3805
                     /note="[] Litmus plasmid::Litmus Sequencing.   Primer for 
                     sequencing Litmus plasmid"
     primer_bind     1183..1203
                     /label=MK 4798
                     /note="[-] GalK::pGalK-seq.#2-3. For sequencing of the 
                     pGalK plasmid in the reverse direction."
     primer_bind     complement(1190..1233)
                     /label=MK 4889
                     /note="[+] AMP NEO BAC::PBAC108L(#1)-Amp-5'. Together with 
                     4890 makes a 2.8 kb fragment containing Amp and NEO 
                     cassettes. Anneals to the Amp site. Use L2neo-IRES-EGFPpA 
                     plasmid from Nigel Killeen as a template. Use the PCR 
                     fragment for the next PCR reaction with primers 4891 x 4892
                     ==> this will add some more homology for BAC108L."
     primer_bind     1248..1269
                     /label=pMAL-c5E ORF.FOR
     misc_feature    1406..1434
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     primer_bind     1432..1452
                     /label=MK 1377
                     /note="[+] pK233::Sequencing/PCR oligo"
     primer_bind     1433..1454
                     /label=MK 1726
                     /note="[+] pTz18R ::5' sequencing oligo for pTZ/pMet"
     primer_bind     1434..1451
                     /label=MK 822
                     /note="[+] pTZ 173-190::reverse primer, upstream of 76"
     primer_bind     1438..1459
                     /label=MK 5558
                     /note="[+] newly made sequence primer 4405::"
     primer_bind     1438..1459
                     /label=MK 4405
                     /note="[+] pET30-BirA::5' Sequencing Oligo"
     primer_bind     1438..1454
                     /label=MK 4291
                     /note="[+] pET-30b vector::sequencing primer - 5' of mult 
                     cloning site of pET-30b (in the lac promoter region)"
     protein_bind    1442..1458
                     /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     complement(1461..1478)
                     /label=MK 1728
                     /note="[-] pTMet::Change Polylinker to add NcoI/BstXI at 5'
                     end"
     CDS             1528..4054
                     /codon_start=1
                     /label=MBP-mRI
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQTNSSSNNNNNNNNNNLGIEGRSLDIQCEQLSDA
                     RWTELLPLIQQYEVVRLDDCGLTEVRCKDISSAVQANPALTELSLRTNELGDGGVGLVL
                     QGLQNPTCKIQKLSLQNCGLTEAGCGILPGMLRSLSTLRELHLNDNPMGDAGLKLLCEG
                     LQDPQCRLEKLQLEYCNLTATSCEPLASVLRVKADFKELVLSNNDLHEPGVRILCQGLK
                     DSACQLESLKLENCGITAANCKDLCDVVASKASLQELDLSSNKLGNAGIAALCPGLLLP
                     SCKLRTLWLWECDITAEGCKDLCRVLRAKQSLKELSLASNELKDEGARLLCESLLEPGC
                     QLESLWIKTCSLTAASCPYFCSVLTKSRSLLELQMSSNPLGDEGVQELCKALSQPDTVL
                     RELWLGDCDVTNSGCSSLANVLLANRSLRELDLSNNCMGGPGVLQLLESLKQPSCTLQQ
                     LVLYDIYWTNEVEEQLRALEEERPSLRIIS"
     primer_bind     1533..1567
                     /label=MK 6773
                     /note="[+] Maltose-binding protein (MBP)::MBP Isolation 
                     Front A.FOR"
     primer_bind     1533..1567
                     /label=MBP Isolation Front A.FOR
     primer_bind     1533..1560
                     /label=MK 6774
                     /note="[+] Maltose-binding protein (MBP)::MBP Isolation 
                     Front B.FOR"
     primer_bind     1533..1560
                     /label=MBP Isolation Front B
     primer_bind     2050..2076
                     /label=MBP Isolation Center A.FOR
     primer_bind     complement(2050..2076)
                     /label=MBP Isolation Center A.REV
     primer_bind     complement(2050..2074)
                     /label=MBP Isolation Center B.REV
     primer_bind     2050..2074
                     /label=MBP Isolation Center Primer B.FOR
     primer_bind     complement(2580..2603)
                     /label=MBP Isolation End B.REV
     primer_bind     complement(2582..2603)
                     /label=MBP Isolation End A.REV
     primer_bind     complement(2585..2613)
                     /label=MK 6795
                     /note="[-] MBP OL-PCR JK::overlap PCR from JK"
     primer_bind     2587..2610
                     /label=MK 2417
                     /note="[+] forward sequence primer for pMAL vector::found 
                     on NEB web site"
     primer_bind     2671..2701
                     /label=MK 6814
                     /note="[+] mouse ribonuclease inhibitor + Fxa Cleavage tag 
                     + Aval site ::56C Tm --> 66C Tm"
     primer_bind     2671..2701
                     /label=mRI cDNA amp2.for
                     /note="56(o)C intially and 66(o)C after amplification"
     primer_bind     2682..2710
                     /label=MK 6812
                     /note="[+] mouse ribonuclease inhibitor + Fxa Cleavage 
                     tag:: 59C tm --> 65C tm"
     primer_bind     2682..2710
                     /label=mRI cDNA amp1.for
                     /note="59(o)C Tm initial and 65(o)C Tm when fully annealed"
     primer_bind     2688..2710
                     /label=MK 6818
                     /note="[+] mouse ribonuclease inhibitor + NdeI::addition of
                     NdeI for cloning into pET19b"
     primer_bind     complement(3263..3285)
                     /label=mRI seq3.rev
     primer_bind     complement(3772..3791)
                     /label=mRI seq2.rev
     primer_bind     complement(4032..4060)
                     /label=MK 6813
                     /note="[-] mouse ribonuclease inhibitor::primer for 
                     sequencing inserted mouse ribonuclease inhibitor into C5x 
                     vector"
     primer_bind     complement(4032..4060)
                     /label=mRI cDNA amp1.rev
     primer_bind     complement(4032..4054)
                     /label=MK 6811
                     /note="[-] mouse ribonuclease inhibitor::primer for 
                     sequencing inserted mouse ribonuclease inhibitor into C5x 
                     vector"
     primer_bind     complement(4037..4059)
                     /label=MK 6819
                     /note="[-] mouse ribonuclease inhibitor + BamHI::addition 
                     of BamHI for cloning into pET19b"
     primer_bind     complement(4038..4066)
                     /label=MK 6815
                     /note="[-] mouse ribonuclease inhibitor::generates a + SbfI
                     site"
     primer_bind     complement(4038..4066)
                     /label=mRI cDNA amp2.rev
     misc_feature    4063..4079
                     /label=MCS
                     /note="multiple cloning site"
     terminator      4080..4166
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4099..4117)
                     /label=T1.rev
                     /note="sequencing primer for RNase inhibitor insert."
     primer_bind     complement(4131..4149)
                     /label=T1_v2.rev
     primer_bind     complement(4226..4251)
                     /label=pMAL-c5E ORF.REV
     terminator      4258..4285
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4313..4403
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4330..4353
                     /label=MK 4294
                     /note="[-] pUC origin, Amp gene::pUC/Amp-ClaI. Together 
                     with 4293, this primer will amplify the pUC-Amp cassette, 
                     which will then be used to construct the DTA-based mIL-15 
                     KIN construct."
     primer_bind     4332..4356
                     /label=MK 4644
                     /note="[-] Kanamycin::Kanamycin (HindIII)-3'. Togetherwith 
                     4643 will amplify the Kanamycin gene which will have an Xba
                     I at the 5' end and HindIII at the 3' end."
     CDS             4404..5264
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4499..4528)
                     /label=MK 4680
                     /note="[-] DTA (Diphteria toxin-A)::DTA ORF#2-3'. 
                     Togetherwith 4665 will amplify the DTA gene cassette from 
                     the IL-15 Knock-IN construct.  This PCR is used to test if 
                     the cassette makes functional gene for negative selection 
                     ==> none of the selected ES clones should have this 
                     cassette intact!"
     primer_bind     complement(4543..4566)
                     /label=MK 4666
                     /note="[-] DTA (Diphteria toxin-A)::DTA ORF#1-3'. 
                     Togetherwith 4665 will amplify the DTA gene cassette from 
                     the IL-15 Knock-IN construct.  This PCR is used to test if 
                     the cassette makes functional gene for negative selection 
                     ==> none of the selected ES clones should have this 
                     cassette intact!"
     primer_bind     complement(5244..5265)
                     /label=MK 4889
                     /note="[+] AMP NEO BAC::PBAC108L(#1)-Amp-5'. Together with 
                     4890 makes a 2.8 kb fragment containing Amp and NEO 
                     cassettes. Anneals to the Amp site. Use L2neo-IRES-EGFPpA 
                     plasmid from Nigel Killeen as a template. Use the PCR 
                     fragment for the next PCR reaction with primers 4891 x 4892
                     ==> this will add some more homology for BAC108L."
     rep_origin      5352..5940
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5996..6017
                     /label=MK 1727
                     /note="[+] pTz18R::From Afl3 site.  Used to make pTMet"
     primer_bind     6019..6039
                     /label=MK 4553
                     /note="[] ::"
     primer_bind     complement(6035..6069)
                     /label=MK 4293
                     /note="[+] pUC origin, Amp gene::pUC/Amp-AvrII. Together 
                     with 4294, this primer will amplify the pUC-Amp cassette, 
                     which will then be used to construct the DTA-based mIL-15 
                     KIN construct."
     primer_bind     6294..6316
                     /label=MK 2420
                     /note="[-] reverse sequence primer for pGEX vector::found 
                     on pharmacia web site"
     CDS             complement(6310..6501)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
       61 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
      121 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
      181 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
      241 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
      301 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
      361 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
      421 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
      481 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
      541 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
      601 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
      661 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
      721 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
      781 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
      841 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatttcggta gtgggatacg
      901 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
      961 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     1021 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     1081 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     1141 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
     1201 gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg
     1261 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     1321 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     1381 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
     1441 attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga
     1501 gcaattgacc aacaaggacc atagattatg aaaatcgaag aaggtaaact ggtaatctgg
     1561 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat
     1621 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt
     1681 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac
     1741 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat
     1801 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt
     1861 gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa
     1921 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac
     1981 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag
     2041 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg
     2101 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac
     2161 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg
     2221 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc
     2281 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt
     2341 ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg
     2401 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag
     2461 ttggtgaaag atccgcgtat tgccgccact atggaaaacg cccagaaagg tgaaatcatg
     2521 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc
     2581 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg
     2641 aacaacaaca acaataacaa taacaacaac ctcgggattg agggaaggag tcttgacatc
     2701 cagtgtgagc agctgagtga tgcccggtgg acagagctcc ttcctctgat ccagcaatac
     2761 gaagtggtca ggctggatga ctgtggcctc actgaagtgc ggtgcaaaga catcagctca
     2821 gcagtccaag ccaaccctgc cctgacagag ctcagccttc gcaccaatga actgggtgat
     2881 ggcggtgtgg gtctggtgct ccagggcctg cagaatccca cttgtaagat ccagaagctg
     2941 agccttcaga actgcggctt gacggaggct ggctgtggga tcctgcctgg catgctgcgc
     3001 tctttgtcta ccctgcgtga gctacatctc aatgataacc ctatggggga cgcaggcctg
     3061 aagctgctct gtgaaggact tcaggacccc cagtgccgtc ttgagaagct tcagttggaa
     3121 tactgtaacc tcacagctac cagctgcgag cccctggctt cagtgctcag ggtgaaagct
     3181 gactttaagg agctagtatt gagcaacaac gacctccatg aacctggtgt ccgtattctg
     3241 tgccagggcc tgaaggactc tgcctgtcaa ctggagtcac tcaagctgga gaactgtggt
     3301 atcacagcag ccaactgcaa ggatctgtgc gatgttgtgg cctccaaagc ttcgctacag
     3361 gaactggact tgagcagcaa caagctaggt aatgcaggca ttgcagcact gtgcccagga
     3421 ctgctgcttc ccagctgtaa gctcaggact ctgtggctct gggaatgtga tatcactgca
     3481 gagggctgca aggacctgtg ccgtgtcctc agagccaagc agagcctgaa ggaactcagc
     3541 cttgcttcca atgagctgaa ggatgagggt gcccggctgc tgtgtgagag cctattagag
     3601 cctggctgtc agctggagtc actgtggata aagacctgta gcctcacagc tgcctcctgt
     3661 ccctacttct gctccgtgtt gaccaaaagc cgttctctgt tggagctgca aatgagcagc
     3721 aacccactgg gagacgaggg agtccaggag ctttgcaagg ccctgagcca gccagataca
     3781 gtgctgcgtg agctttggtt gggagactgt gatgtgacaa atagtggttg cagcagcctt
     3841 gccaatgtcc tgctggccaa ccgcagcctg agggaactgg acctcagtaa caactgcatg
     3901 gggggcccag gtgtcctgca actgctggag agcctcaaac agcccagctg cacccttcag
     3961 cagctcgtcc tgtatgacat ttactggaca aatgaggtgg aagaacagct tcgggccctg
     4021 gaggaggaaa ggccttccct gaggatcatt tcctaacctg caggtaatta aataagcttc
     4081 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     4141 tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac
     4201 ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga
     4261 aggccatcct gacggatggc ctttttgcgt ttctacaaac tctttcggtc cgttgtttat
     4321 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4381 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4441 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4501 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4561 agatccttga gagttttcgc cccgaagaac gtttcccaat gatgagcact tttaaagttc
     4621 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     4681 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4741 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4801 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4861 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4921 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4981 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5041 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5101 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5161 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5221 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5281 actcatatat actttagatt gatttcctta ggactgagcg tcaaccccgt agaaaagatc
     5341 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5401 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5461 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     5521 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5581 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5641 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5701 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5761 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5821 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5881 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5941 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6001 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     6061 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     6121 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatataa
     6181 ggtgcactgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     6241 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     6301 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     6361 ctcatcagcg tggtcgtgca gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     6421 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     6481 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     6541 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     6601 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     6661 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     6721 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     6781 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     6841 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     6901 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
     6961 cacccgtggc caggacccaa cgctgcccga aatt
//
