LOCUS       sequence-467820-       11611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-inducible mitochondria-localized ApaLI expressing 
            plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11611)
  AUTHORS   Fu Y, Land M, Cui R, Kavlashvili T, Kim M, Lieber T, Ryu KW, 
            DeBitetto E, Masilionis I, Saha R, Takizawa M, Baker D, Tigano M, 
            Reznik E, Sharma R, Chaligne R, Thompson CB, Pe'er D, Sfeir A
  TITLE     Engineering mtDNA Deletions by Reconstituting End-Joining in Human 
            Mitochondria.
  JOURNAL   bioRxiv [Preprint]. 2024 Oct 17:2024.10.15.618543. doi: 
            10.1101/2024.10.15.618543.
  PUBMED    39463974
REFERENCE   2  (bases 1 to 11611)
  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: "bioRxiv 
            [Preprint]. 2024 Oct 17:2024.10.15.618543. doi: 
            10.1101/2024.10.15.618543."
COMMENT     Sequence Label: TLCV2-ApaLI-T2A-EGFP-Puro
FEATURES             Location/Qualifiers
     source          1..11611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        46..360
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    54..72
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    90..108
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    125..143
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    161..179
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    197..215
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    232..250
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    268..286
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     333..357
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             375..449
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1608..1631
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1650..1703
                     /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             join(1704..1706,1707..2420)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(1704..1706,1707..2417)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1746..1767)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2007..2026)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2354..2375
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        2431..2642
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2664..3260
                     /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(2664..2683)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3160..3180
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             3270..3326
                     /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(3327..3944,3945..3950,3951..3986,3987..3989,3990..4025,
                     4026..4028,4029..4064,4065..4073)
                     /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    4094..4682
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4147..4167)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4565..4576)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4754..4987
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5013..5030)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5019..5243
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5289..5717
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5376..5395)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5586..5607
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5726..5746)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5731..6060
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5911..6046
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5973..5992
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6108..6155
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6174..6548
                     /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    6678..6799
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6715..6734)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6769..6788
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6848..6864)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6848..6864)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6861..6883)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6872..6888
                     /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(6896..6902,6903..6920,6921..6926))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6941..6962
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7079..7096)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7250..7838)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7330..7349)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8009..8800,8801..8869))
                     /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     8632..8651
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8870..8974)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9049..9068)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        9240..9619
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9621..9819
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             9837..10017
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    10064..10189
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10682..10915
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11100..11144
                     /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    11442..11559
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gggcctattt cttttgaatt cactttggcc gcgaatcgat atgtcgagtt tactccctat
       61 cagtgataga gaacgtatgt cgagtttact ccctatcagt gatagagaac gatgtcgagt
      121 ttactcccta tcagtgatag agaacgtatg tcgagtttac tccctatcag tgatagagaa
      181 cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt atccctatca
      241 gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacgt atgtcgaggt
      301 aggcgtgtac ggtgggaggc ctatataagc agagctcgtt tagtgaaccg tcagatcgcc
      361 tggagaatac cggtatgtcc gtcctgacgc cgctgctgct gcggggcttg acaggctcgg
      421 cccggcggct cccagtgccg cgcgccaaga tccattcgtt gccgccggag gggaagctta
      481 tgaccacacg acagcgctta tctgctgaga gatctcagca gcttacccgt ctgctcacta
      541 tcactaaaac cgctaacatg cgggccttaa tggaggctag cgaactcgct aaagtaattg
      601 ccctggtggc ggttgacata ggtaagtcag acgagatggc cagagctttc cccgtgctgt
      661 ggccaaaaat cagtcctcag caagaatatt acgcaacagc ggttgactgg ttcaccaacc
      721 ccgatgaaac cgtcacctct ttcgatgtgg tggatatgct agatgccggc acttccctgg
      781 accaagactt catgacttat ctgaaatgtc taactgaact gcacaagcga cggagaaaat
      841 acggcctgat cctgcagagg cagcctctac caacgatggt gcaggtttcg ccccgagcac
      901 tgatggaata cggacctgac tttccgcccg aggcactggc tagctggctt acctggcgga
      961 agttttttta cgaccttgac aacaggtctg cccaggaaac agggtacttg ttcgaaccca
     1021 ttctggccgc cgccatcgga ggcgaggcaa aatctgccag agagcgtgtc gtaagacgca
     1081 ccgacgatcc aacgaagggc aggcaggtgg attgctggaa ggtgctccct gatggtacgc
     1141 ctctggctta tgaattgaaa cttcgggtga caattgcagc gagcgggcaa ggcagatttg
     1201 gagaggagct gagcttcgct cgggactgca gcagctccgg cgctaagccg atcctcgtgg
     1261 tcctggatcc aacagaaaat gacaagctca ctggattgca agccgcatac cgcgaggttg
     1321 gaggcgccgc atatgtgggg gatgccgcct gggctcactt agaggacgag gctggtgcta
     1381 ccatggccag ttttatagag cgctatgtca gggtgccagt agcatcagtc agttccttcg
     1441 aaagggtcat tgagggagat gccacaaagc gctccttgat cctgcaggac ctccaggccc
     1501 gactcgatgg taatgagctt acaatctccc tgggaggcca tcagcggttg gtggaaaggc
     1561 atgaggacca gagtctcgca gcggacggcg atgacgattc agagctcgat tacaaagacg
     1621 atgacgataa gggcgcgcct ggatccggag agggcagagg aagtctgcta acatgcggtg
     1681 acgtcgagga gaatcctggc ccagtgagca agggcgagga gctgttcacc ggggtggtgc
     1741 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1801 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     1861 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     1921 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1981 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     2041 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     2101 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     2161 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     2221 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2281 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     2341 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2401 tggacgagct gtacaagtaa gctagcatcc gggcagagcg cacatcgccc acagtccccg
     2461 agaagttggg gggaggggtc ggcaattgat ccggtgccta gagaaggtgg cgcggggtaa
     2521 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
     2581 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
     2641 agactgcgat cgcaatgtac agtatgaccg agtacaagcc cacggtgcgc ctcgccaccc
     2701 gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
     2761 cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct
     2821 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
     2881 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
     2941 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
     3001 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcgga gtctcgcccg
     3061 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     3121 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
     3181 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
     3241 tgacccgcaa gcccggtgcc ggttccggcg caacaaactt ctctctgctg aaacaagccg
     3301 gagatgtcga agagaatcct ggaccgatgt ctagactgga caagagcaaa gtcataaacg
     3361 gagctctgga attactcaat ggtgtcggta tcgaaggcct gacgacaagg aaactcgctc
     3421 aaaagctggg agttgagcag cctaccctgt actggcacgt gaagaacaag cgggccctgc
     3481 tcgatgccct gccaatcgag atgctggaca ggcatcatac ccacttctgc cccctggaag
     3541 gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc ataccgctgt gctctcctct
     3601 cacatcgcga cggggctaaa gtgcatctcg gcacccgccc aacagagaaa cagtacgaaa
     3661 ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt ctccctggag aacgcactgt
     3721 acgctctgtc cgccgtgggc cactttacac tgggctgcgt attggaggaa caggagcatc
     3781 aagtagcaaa agaggaaaga gagacaccta ccaccgattc tatgccccca cttctgagac
     3841 aagcaattga gctgttcgac cggcagggag ccgaacctgc cttccttttc ggcctggaac
     3901 taatcatatg tggcctggag aaacagctaa agtgcgaaag cggcgggccg accgacgccc
     3961 ttgacgattt tgacttagac atgctcccag ccgatgccct tgacgacttt gaccttgata
     4021 tgctgcctgc tgacgctctt gacgattttg accttgacat gctccccggg taactcgagc
     4081 gcgttaagtc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     4141 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     4201 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     4261 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     4321 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     4381 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     4441 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     4501 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     4561 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     4621 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     4681 gcgtcgactt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     4741 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
     4801 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     4861 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     4921 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     4981 tctagcaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     5041 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     5101 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     5161 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     5221 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     5281 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     5341 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     5401 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     5461 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     5521 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5581 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     5641 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     5701 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     5761 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     5821 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5881 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5941 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     6001 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     6061 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat
     6121 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca
     6181 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
     6241 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
     6301 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
     6361 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
     6421 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
     6481 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
     6541 aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     6601 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     6661 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     6721 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     6781 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     6841 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6901 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6961 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     7021 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     7081 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     7141 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     7201 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     7261 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     7321 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     7381 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     7441 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7501 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7561 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7621 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7681 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7741 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7801 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7861 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7921 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7981 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     8041 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     8101 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     8161 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     8221 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     8281 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     8341 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     8401 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     8461 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     8521 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     8581 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     8641 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     8701 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     8761 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8821 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8881 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8941 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     9001 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
     9061 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     9121 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     9181 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
     9241 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     9301 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     9361 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     9421 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     9481 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     9541 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     9601 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     9661 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     9721 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     9781 gggcggtagg cgtgtacggt gggaggtcta tataagcagc gcgttttgcc tgtactgggt
     9841 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     9901 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     9961 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
    10021 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
    10081 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
    10141 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
    10201 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
    10261 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
    10321 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
    10381 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
    10441 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
    10501 caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata
    10561 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
    10621 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
    10681 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
    10741 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
    10801 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
    10861 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
    10921 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
    10981 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
    11041 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
    11101 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
    11161 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
    11221 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
    11281 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag
    11341 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tcggcactgc
    11401 gtgcgccaat tctgcagaca aatggcagta ttcatccaca attttaaaag aaaagggggg
    11461 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
    11521 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
    11581 agagatccag tttggttaat taaggtaccg a
//
