LOCUS       sequence-345296-        9841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optogenetic coupling of RAB11 to tetramerized moss kinesin-14 to 
            induce retrograde transport of recycling endosomes. Compatible with 
            Flp-in TREX system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9841)
  AUTHORS   Nijenhuis W, van Grinsven MMP, Kapitein LC
  TITLE     An optimized toolbox for the optogenetic control of intracellular 
            transport.
  JOURNAL   J Cell Biol. 2020 Apr 6;219(4). pii: 133834. doi: 
            10.1083/jcb.201907149.
  PUBMED    32328628
REFERENCE   2  (bases 1 to 9841)
  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: "J Cell 
            Biol."; date: "2020-04-6"; volume: "219(4). pii: 133834. doi"; 
            pages: " 10.1083/jcb.201907149"
COMMENT     Sequence Label: 
            pCDNA5-FRT-TO-SSPB(micro)-mVenus-GCN4-ppKin14VIb(861-1321)_P2A_iLID-
            mCherry-RAB11
FEATURES             Location/Qualifiers
     source          1..9841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(252..254,255..965)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(252..254,255..965)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(294..315)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(555..574)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     902..923
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    975..999
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     RBS             1463..1471
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             2550..2606
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2613..3050
                     /codon_start=1
                     /product="improved light-inducible dimer when used with 
                     SspB nano or SspB micro (Guntas et al., 2015)"
                     /label=iLID
                     /translation="LATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNVFHLQPMRDYKGDVQ
                     YFIGVQLDGTERLHGAAEREAVCLIKKTAFQIAEAANDENYF"
     CDS             3090..3794
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(3232..3250)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(4573..4590)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4579..4803
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4936..4955)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5087..5134
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             5142..6164
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             5142..6164
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    6294..6415
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6331..6350)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6385..6404
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6464..6480)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6464..6480)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6477..6499)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6488..6504
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6512..6518,6519..6536,6537..6542))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6557..6578
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6695..6712)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6866..7454)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6946..6965)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7625..8416,8417..8485))
                     /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     8248..8267
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8486..8590)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8665..8684)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        8856..9235
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9236..9439
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9390..9410
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    9441..9459
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9462..9480
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     9490..9514
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      9616..9625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(9655..9841,1..20,21..23,24..143)
                     /codon_start=1
                     /product="SspB from Haemophilus influenzae, with the 
                     mutations Y11K and A15E to disrupt the natural dimerization
                     of the protein and with the mutation R74Q to reduce the 
                     affinity for iLID"
                     /label=SspB micro
                     /note="dimerizes with iLID at moderate affinity to create 
                     iLID micro (Guntas et al., 2015)"
                     /translation="SSPKRPKLLREYYDWLVDNSFTPYLVVDATYLGVNVPVEYVKDGQ
                     IVLNLSASATGNLQLTNDFIQFNAQFKGVSRELYIPMGAALAIYARENGDGVMFEPEEI
                     YDELNI"
ORIGIN
        1 attttatcca gttcaacgcc cagtttaagg gcgtgtctcg tgaactgtat atcccgatgg
       61 gtgccgctct ggccatttac gctcgcgaga acggcgatgg tgtgatgttc gaaccagaag
      121 aaatctatga cgagctgaat attggtcaat tgtctagagg tggcgggggc agtgggggag
      181 ggggaagtgg aggcggcggt tctggcgggg gaggctccgg tggcgggggg agcgggggtg
      241 ggagtaagct tgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      301 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      361 ccacctacgg caagctgacc ctgaagttga tctgcaccac cggcaagctg cccgtgccct
      421 ggcccaccct cgtgaccacc ctcggctacg gcctgcagtg cttcgcccgc taccccgacc
      481 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      541 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      601 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      661 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
      721 agaagaacgg catcaaggca aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
      781 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      841 acaaccacta cctgagctac cagtccaaac tgagcaaaga ccccaacgag aagcgcgatc
      901 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
      961 acaagcaccg gtatacaagt ttgtacaaaa aagcaggctc cgctattgaa gacaagttgg
     1021 aagaaatact ttcgaaattg taccacattg aaaatgagtt ggccagaatt aagaaattat
     1081 tgggcgaaat tggtggaggt agtggtggag gtagtgcggc cgccagagat gatttgaggg
     1141 gacgacttgt aactctcgag aagaaattag cagagcgtac ccaagatttg agtcttgcag
     1201 agtctacttt ggcaactcgt gagggtgaat tcgaatccct gcaaagcaat ctaatggagc
     1261 ttaatgagct ccgggagatg aaggaggata tcgacagaaa gaatgagcaa acagcagcta
     1321 tattgaagcg acaagcagac cagataactg aattagaaac tctttatagg gaagaacaag
     1381 tccttcgaaa acgatacttc aacatgatgg aagacatgaa aggtaagatc agggtatatg
     1441 ctcggtggag acccctaagt agtaaggagg tcaaagagcg gcaacaaaac gtgttgatag
     1501 cacctgatga attcaccatc gaacatcctt ggaaagacga caaacccaaa caacatcaat
     1561 ttgatcacgt tttcgaccac catgcgaccc aagaagaggt ctttgaggat acaaagtatt
     1621 tggtccaatc agcaattgac ggatacaatg tctgtatatt tgcatacggg caaactgggt
     1681 caggcaaaac gtttacgatt tacggttctg acaacaaccc aggtctaact ccacgagcta
     1741 ccaaagaact ttttggctat cttaagcgag acgccaataa gttttcgttt gctctgaagg
     1801 tttacatgct tgaaatatat caagattcgc tcatcgatct gcttctacca aagagtgctg
     1861 caaagccacg aaagcttgag attaagaagg actccaaggg aatggttgtc gtagagaatg
     1921 caactctact gccgattgca tcccatgatg aactgcaagc cattgttcat aagggtctgg
     1981 agagacgaca tgtttcgggc acccacatga atgcggagag ttcacggtca cacctcattc
     2041 tatcagtaat cgttgaaagc accaaccgac agtctcaagt tcttgtgaag ggcaagctga
     2101 gttttgtgga tcttgcgggc tctgaaagag tgaagaaatc tggatccagt ggtgaacaac
     2161 tgaaagaggc ccaaagcatt aataaatctt tgtcagcact aggtgatgtc attagcgccc
     2221 ttgcaacaga ggagcagcat attccttacc gtaaccacaa gctaacaatg cttatgagcg
     2281 attctctcgg tggaaatgcg aagaccctca tgtttgtaaa tatctcacca gcagagtcta
     2341 atctggacga gacccataat tcactctgct atgccacaag ggtaagatcc atcatcaatg
     2401 aggccagtaa aaatactaca acgaaagaga tactgcgact gaagaaacaa atcgcgtact
     2461 ggaaggagca agcgggcagg atgggagggt ccgaggatct ggaagaggtt atggatgatc
     2521 gtggaatcaa aactgcagat ggatccggag ccacgaactt ctctctgtta aagcaagcag
     2581 gagacgtgga agaaaacccc gggcccacca tgctggcaac cacactggaa cggatcgaga
     2641 aaaatttcgt gattactgat ccgagactgc ctgacaaccc aatcattttt gcgagcgatt
     2701 ccttcctgca gctgacagaa tattctcggg aagagatcct ggggcgcaat tgccgttttc
     2761 tgcagggacc cgagacagac cgtgccactg ttcggaaaat cagagatgct attgacaacc
     2821 agactgaagt gaccgttcag ctgatcaatt ataccaagag cggcaagaag ttctggaacg
     2881 tgttccacct gcagccgatg cgcgattata agggcgacgt ccagtacttc attggcgtgc
     2941 agctggatgg caccgaacgt cttcatggcg ccgctgagcg tgaggcggtc tgcctgatca
     3001 aaaagacagc ctttcagatt gctgaggcag cgaacgacga aaattacttt tctagaggtg
     3061 gcggggggag cgggggtggg agtaagcttg tgagcaaggg cgaggaggat aacatggcca
     3121 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
     3181 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     3241 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
     3301 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
     3361 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     3421 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     3481 ccaacttccc ctccgacggt cccgtaatgc agaagaagac catgggctgg gaggcctcct
     3541 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     3601 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     3661 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     3721 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     3781 acgagctgta caaggctagc ggaggcggtg gctcaggcgg aggagggtca ggagggggag
     3841 gcagtggcgg gggcggctct ggaggagggg gttctggtgg tggtagctac gtaggcaccc
     3901 gcgacgacga gtacgactat ctcttcaaag ttgtccttat tggagattct ggtgttggaa
     3961 agagtaatct cttgtctcga tttactcgaa atgagtttaa tctcgaaagt aagagcacca
     4021 ttggagtaga gtttgcaaca agaagcatcc aggttgatgg gaaaacaata aaggcacaga
     4081 tatgggacac ggcagggcag gagcgatacc gagctataac atcagcatat tatcgtggag
     4141 ctgtaggtgc cttactggtt tatgacattg ctaagcatct cacatatgaa aatgtagaac
     4201 gatggctgaa agaactgaga gatcatgctg atagtaacat tgttatcatg cttgtgggca
     4261 ataagagtga tttgcgtcat ctcagggcag ttcctacaga tgaagcaaga gcttttgcag
     4321 aaaagaatgg tttgtcattc attgagactt ccgctctaga ctctacaaat gtagaagctg
     4381 cttttcagac aattctgaca gagatatacc gcattgtttc ccagaaacaa atgtcagaca
     4441 gacgtgaaaa tgacatgtct ccaagcaaca atgtggttcc tattcatgtt ccaccaacca
     4501 ctgaaaacaa gccaaaggta caatgttgtc agaacatcta aaccggtggg cccgtttaaa
     4561 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     4621 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     4681 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     4741 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     4801 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     4861 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4921 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4981 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5041 acctcgaccc caaaaaactt gattagggtg atggttcacg tacctagaag ttcctattcc
     5101 gaagttccta ttctctagaa agtataggaa cttccttggc caaaaagcct gaactcaccg
     5161 cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc
     5221 tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc
     5281 tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg
     5341 catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc gagagcctga
     5401 cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac
     5461 tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta
     5521 gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc
     5581 gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg
     5641 acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact
     5701 gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca
     5761 atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg
     5821 aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct
     5881 acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc
     5941 gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt
     6001 gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac
     6061 aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata
     6121 gtggaaaccg acgccccagc actcgtccga gggcaaagga atagcacgta ctacgagatt
     6181 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     6241 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     6301 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6361 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6421 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     6481 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6541 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6601 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6661 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6721 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6781 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6841 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6901 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6961 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     7021 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     7081 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     7141 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     7201 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     7261 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     7321 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     7381 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7441 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7501 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7561 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7621 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7681 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7741 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7801 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7861 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7921 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7981 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     8041 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     8101 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     8161 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     8221 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     8281 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     8341 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8401 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8461 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8521 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8581 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     8641 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     8701 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     8761 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     8821 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
     8881 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     8941 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     9001 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     9061 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     9121 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     9181 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     9241 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     9301 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     9361 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     9421 ggtctatata agcagagctc tccctatcag tgatagagat ctccctatca gtgatagaga
     9481 tcgtcgacga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt
     9541 tgacctccat agaagacacc gggaccgatc cagcctccgg actctagcgt ttaaacttaa
     9601 gcttgccacc ggcgcgccac catggatctg agcggcctga ccctgcagga attcagctcc
     9661 ccgaaacgcc ctaagctgct gcgtgaatat tacgattggc tggttgataa cagctttacc
     9721 ccatatctgg tggtggatgc cacatacctg ggcgtgaacg tgcccgtgga gtatgtgaaa
     9781 gacggtcaga tcgtgctgaa tctgtctgca agtgcgaccg gcaacctgca actgacaaat
     9841 g
//
