LOCUS       sequence-417768-       11626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus backbone expressing a doxycycline-inducible mitochondrial
            localized ApaLI mutant (D211A, E225A and K227A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11626)
  AUTHORS   Fu Y, Sacco O, DeBitetto E, Kanshin E, Ueberheide B, Sfeir A
  TITLE     Mitochondrial DNA breaks activate an integrated stress response to 
            reestablish homeostasis.
  JOURNAL   Mol Cell. 2023 Oct 8:S1097-2765(23)00753-0. doi: 
            10.1016/j.molcel.2023.09.026.
  PUBMED    37832546
REFERENCE   2  (bases 1 to 11626)
  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: "Mol Cell. 
            2023 Oct 8:S1097-2765(23)00753-0. doi: 
            10.1016/j.molcel.2023.09.026."
COMMENT     Sequence Label: TLCV2-ApaLI*-FLAG-T2A-mTagBFP2-Puro
FEATURES             Location/Qualifiers
     source          1..11626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(360..371)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             549..782
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(808..825)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    814..1038
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1084..1512
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1171..1190)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1381..1402
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1521..1541)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1526..1855
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1706..1841
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1768..1787
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1903..1950
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1969..2343
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    2473..2594
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2510..2529)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2564..2583
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2643..2659)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2643..2659)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2656..2678)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2667..2683
                     /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(2691..2697,2698..2715,2716..2721))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2736..2757
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2874..2891)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3045..3633)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3125..3144)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3804..4595,4596..4664))
                     /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     4427..4446
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4665..4769)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4844..4863)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5035..5414
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5416..5614
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             5632..5812
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5859..5984
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6477..6710
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6895..6939
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7237..7354
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7452..7766
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    7460..7478
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7496..7514
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7531..7549
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7567..7585
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7603..7621
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7638..7656
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7674..7692
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     7739..7763
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             7781..7855
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             9014..9037
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9062..9115
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             9134..9829
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="ELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVVE
                     GGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTATQ
                     DTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKLV
                     GGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCDL
                     PSKLGHKLN"
     promoter        9852..10063
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             10085..10681
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(10085..10104)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     10581..10601
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             10691..10747
                     /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             join(10748..11365,11366..11371,11372..11407,11408..11410,
                     11411..11446,11447..11449,11450..11485,11486..11494)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    join(11515..11626,1..477)
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11568..11588)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
       61 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
      121 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
      181 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
      241 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
      301 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
      361 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
      421 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcgtc
      481 gactttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
      541 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
      601 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
      661 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
      721 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
      781 cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
      841 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
      901 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
      961 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     1021 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     1081 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     1141 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     1201 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     1261 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     1321 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     1381 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     1441 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     1501 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     1561 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     1621 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     1681 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     1741 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     1801 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     1861 cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg
     1921 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg
     1981 accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc
     2041 gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac
     2101 gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc
     2161 tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg
     2221 aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg
     2281 gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac
     2341 tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     2401 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     2461 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2521 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2581 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     2641 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     2701 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     2761 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     2821 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     2881 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2941 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3001 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3061 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3121 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3181 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3241 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3301 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3361 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3421 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3481 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3541 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3601 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3661 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3721 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3781 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3841 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3901 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3961 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4021 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4081 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4141 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4201 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4261 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4321 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4381 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4441 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4501 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4561 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4621 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4681 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4741 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     4801 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     4861 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     4921 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     4981 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
     5041 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     5101 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     5161 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     5221 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     5281 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     5341 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     5401 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     5461 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     5521 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     5581 gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc
     5641 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     5701 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     5761 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     5821 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     5881 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     5941 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     6001 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     6061 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     6121 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     6181 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     6241 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     6301 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
     6361 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     6421 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     6481 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     6541 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     6601 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     6661 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     6721 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     6781 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     6841 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     6901 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     6961 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     7021 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     7081 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
     7141 ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg
     7201 ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
     7261 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
     7321 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga
     7381 tccagtttgg ttaattaagg taccgagggc ctatttcttt tgaattcact ttggccgcga
     7441 atcgatatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag tttactccct
     7501 atcagtgata gagaacgatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga
     7561 gtttactccc tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag
     7621 aacgtatgtc gagtttatcc ctatcagtga tagagaacgt atgtcgagtt tactccctat
     7681 cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg ggaggcctat ataagcagag
     7741 ctcgtttagt gaaccgtcag atcgcctgga gaataccggt atgtccgtcc tgacgccgct
     7801 gctgctgcgg ggcttgacag gctcggcccg gcggctccca gtgccgcgcg ccaagatcca
     7861 ttcgttgccg ccggagggga agcttatgac cacacgacag cgcttatctg ctgagagatc
     7921 tcagcagctt acccgtctgc tcactatcac taaaaccgct aacatgcggg ccttaatgga
     7981 ggctagcgaa ctcgctaaag taattgccct ggtggcggtt gacataggta agtcagacga
     8041 gatggccaga gctttccccg tgctgtggcc aaaaatcagt cctcagcaag aatattacgc
     8101 aacagcggtt gactggttca ccaaccccga tgaaaccgtc acctctttcg atgtggtgga
     8161 tatgctagat gccggcactt ccctggacca agacttcatg acttatctga aatgtctaac
     8221 tgaactgcac aagcgacgga gaaaatacgg cctgatcctg cagaggcagc ctctaccaac
     8281 gatggtgcag gtttcgcccc gagcactgat ggaatacgga cctgactttc cgcccgaggc
     8341 actggctagc tggcttacct ggcggaagtt tttttacgac cttgacaaca ggtctgccca
     8401 ggaaacaggg tacttgttcg aacccattct ggccgccgcc atcggaggcg aggcaaaatc
     8461 tgccagagag cgtgtcgtaa gacgcaccga cgatccaacg aagggcaggc aggtggcctg
     8521 ctggaaggtg ctccctgatg gtacgcctct ggcttatgcg ttggcgcttc gggtgacaat
     8581 tgcagcgagc gggcaaggca gatttggaga ggagctgagc ttcgctcggg actgcagcag
     8641 ctccggcgct aagccgatcc tcgtggtcct ggatccaaca gaaaatgaca agctcactgg
     8701 attgcaagcc gcataccgcg aggttggagg cgccgcatat gtgggggatg ccgcctgggc
     8761 tcacttagag gacgaggctg gtgctaccat ggccagtttt atagagcgct atgtcagggt
     8821 gccagtagca tcagtcagtt ccttcgaaag ggtcattgag ggagatgcca caaagcgctc
     8881 cttgatcctg caggacctcc aggcccgact cgatggtaat gagcttacaa tctccctggg
     8941 aggccatcag cggttggtgg aaaggcatga ggaccagagt ctcgcagcgg acggcgatga
     9001 cgattcagag ctcgattaca aagacgatga cgataagggc gcgcctggat ccggcagtgg
     9061 agagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccaatggt
     9121 gagcaagggt gaggagctga ttaaggagaa catgcacatg aagctgtaca tggagggcac
     9181 cgtggacaac catcacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac
     9241 ccagaccatg agaatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct
     9301 ggctactagc ttcctctacg gcagcaagac cttcatcaac cacacccagg gcatccccga
     9361 cttcttcaag cagtccttcc ctgagggctt cacatgggag agagtcacca catacgaaga
     9421 cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc tcatctacaa
     9481 cgtcaagatc agaggggtga acttcacatc caacggccct gtgatgcaga agaaaacact
     9541 cggctgggag gccttcaccg agacgctgta ccccgctgac ggcggcctgg aaggcagaaa
     9601 cgacatggcc ctgaagctcg tgggcgggag ccatctgatc gcaaacgcca agaccacata
     9661 tagatccaag aaacccgcta agaacctcaa gatgcctggc gtctactatg tggactacag
     9721 actggaaaga atcaaggagg ccaacaacga gacctacgtc gagcagcacg aggtggcagt
     9781 ggccagatac tgcgacctcc ctagcaaact ggggcacaag cttaattgac cattcgattg
     9841 ggctagcatc cgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
     9901 cggcaattga tccggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
     9961 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
    10021 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagactgcga tcgcaatgta
    10081 cagtatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
    10141 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
    10201 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
    10261 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
    10321 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
    10381 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
    10441 ggagcccgcg tggttcctgg ccaccgtcgg agtctcgccc gaccaccagg gcaagggtct
    10501 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
    10561 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
    10621 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
    10681 cggttccggc gcaacaaact tctctctgct gaaacaagcc ggagatgtcg aagagaatcc
    10741 tggaccgatg tctagactgg acaagagcaa agtcataaac ggagctctgg aattactcaa
    10801 tggtgtcggt atcgaaggcc tgacgacaag gaaactcgct caaaagctgg gagttgagca
    10861 gcctaccctg tactggcacg tgaagaacaa gcgggccctg ctcgatgccc tgccaatcga
    10921 gatgctggac aggcatcata cccacttctg ccccctggaa ggcgagtcat ggcaagactt
    10981 tctgcggaac aacgccaagt cataccgctg tgctctcctc tcacatcgcg acggggctaa
    11041 agtgcatctc ggcacccgcc caacagagaa acagtacgaa accctggaaa atcagctcgc
    11101 gttcctgtgt cagcaaggct tctccctgga gaacgcactg tacgctctgt ccgccgtggg
    11161 ccactttaca ctgggctgcg tattggagga acaggagcat caagtagcaa aagaggaaag
    11221 agagacacct accaccgatt ctatgccccc acttctgaga caagcaattg agctgttcga
    11281 ccggcaggga gccgaacctg ccttcctttt cggcctggaa ctaatcatat gtggcctgga
    11341 gaaacagcta aagtgcgaaa gcggcgggcc gaccgacgcc cttgacgatt ttgacttaga
    11401 catgctccca gccgatgccc ttgacgactt tgaccttgat atgctgcctg ctgacgctct
    11461 tgacgatttt gaccttgaca tgctccccgg gtaactcgag cgcgttaagt cgacaatcaa
    11521 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
    11581 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattg
//
