LOCUS       sequence-467822-       11223 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Doxycycline-inducible mitochondria-localized ScaI expressing 
            plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11223)
  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 11223)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-Scal-T2A-EGFP-Puro
FEATURES             Location/Qualifiers
     source          1..11223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        59..373
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    67..85
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    103..121
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    138..156
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    174..192
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    210..228
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    245..263
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    281..299
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     346..370
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             396..470
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1242..1265
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1275..1328
                     /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(1329..1331,1332..2045)
                     /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(1329..1331,1332..2042)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1371..1392)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1632..1651)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1979..2000
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        2056..2267
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2289..2885
                     /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(2289..2308)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2785..2805
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2895..2951
                     /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(2952..3569,3570..3575,3576..3611,3612..3614,3615..3650,
                     3651..3653,3654..3689,3690..3698)
                     /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    3719..4307
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3772..3792)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4190..4201)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4379..4612
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(4638..4655)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4644..4868
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4914..5342
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5001..5020)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5211..5232
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5351..5371)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5356..5685
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5536..5671
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5598..5617
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5733..5780
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5799..6173
                     /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    6303..6424
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6340..6359)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6394..6413
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6473..6489)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6473..6489)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6486..6508)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6497..6513
                     /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(6521..6527,6528..6545,6546..6551))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6566..6587
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6704..6721)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6875..7463)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6955..6974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7634..8425,8426..8494))
                     /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     8257..8276
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8495..8599)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8674..8693)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        8865..9244
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9246..9444
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             9462..9642
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9689..9814
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10307..10540
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10725..10769
                     /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    11067..11184
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 attaaggtac cgagggccta tttcttttga attcactttg gccgcgaatc gatatgtcga
       61 gtttactccc tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag
      121 aacgatgtcg agtttactcc ctatcagtga tagagaacgt atgtcgagtt tactccctat
      181 cagtgataga gaacgtatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag
      241 tttatcccta tcagtgatag agaacgtatg tcgagtttac tccctatcag tgatagagaa
      301 cgtatgtcga ggtaggcgtg tacggtggga ggcctatata agcagagctc gtttagtgaa
      361 ccgtcagatc gcctggagaa taccggtgtg ccatcatgtc cgtcctgacg ccgctgctgc
      421 tgcggggctt gacaggctcg gcccggcggc tcccagtgcc gcgcgccaag atccattcgt
      481 tgccgccgga ggggaagctt atgatcaacg atcagcttcc ccggtgggtg cgcgaggcgc
      541 gcgtgggtac aagaacaggc ggccctgcaa tgcgcccgaa aacttccgat tcgccctact
      601 ttggatggga tagcgaagat tggcccgaag tgactcgcca gctactgtcc gagcaaccgc
      661 tcagcggcga cacgcttgtc gatgccgtgc tcgcttcgtg ggaatcgatc ttcgaatccc
      721 ggctggggag cggatttcac atcggcaccc agatccggcc gactccacaa ataatgggat
      781 tcctccttca tgcactcatc cctcttgagc tcgccaacgg cgacccgagc tggcgcgcgg
      841 acctgaattc ctccgaaaag gatctcgtgt atcagccgga tcataaatat tcaatcgaga
      901 tgaaaacctc gtcacacaag gatcaaattt tcggcaaccg gagcttcgga gtcgagaatc
      961 ccggcaaggg aaagaaggca aaggacggct attatgtggc ggtaaacttc gaaaaatgga
     1021 gtgacgctcc gggcagactt ccacgcatcc ggacgatccg ctatgggtgg cttgaccata
     1081 cagattgggt ggcgcagaaa tctcaaacgg gccagcagtc gtcactaccg gccgtcgtat
     1141 ccaacactca actcctcgcc atccatacgg gtggccagag gtaccaaggg cgaattctgc
     1201 agatatccat cacactggcg gccgctcgag catgcatctc tgattacaaa gacgatgacg
     1261 ataagggatc cggagagggc agaggaagtc tgctaacatg cggtgacgtc gaggagaatc
     1321 ctggcccagt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1381 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     1441 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1501 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     1561 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1621 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1681 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1741 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1801 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1861 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1921 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1981 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     2041 agtaagctag catccgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     2101 gggtcggcaa ttgatccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     2161 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     2221 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagact gcgatcgcaa
     2281 tgtacagtat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca
     2341 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
     2401 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
     2461 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
     2521 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
     2581 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
     2641 ccaaggagcc cgcgtggttc ctggccaccg tcggagtctc gcccgaccac cagggcaagg
     2701 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
     2761 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
     2821 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
     2881 gtgccggttc cggcgcaaca aacttctctc tgctgaaaca agccggagat gtcgaagaga
     2941 atcctggacc gatgtctaga ctggacaaga gcaaagtcat aaacggagct ctggaattac
     3001 tcaatggtgt cggtatcgaa ggcctgacga caaggaaact cgctcaaaag ctgggagttg
     3061 agcagcctac cctgtactgg cacgtgaaga acaagcgggc cctgctcgat gccctgccaa
     3121 tcgagatgct ggacaggcat catacccact tctgccccct ggaaggcgag tcatggcaag
     3181 actttctgcg gaacaacgcc aagtcatacc gctgtgctct cctctcacat cgcgacgggg
     3241 ctaaagtgca tctcggcacc cgcccaacag agaaacagta cgaaaccctg gaaaatcagc
     3301 tcgcgttcct gtgtcagcaa ggcttctccc tggagaacgc actgtacgct ctgtccgccg
     3361 tgggccactt tacactgggc tgcgtattgg aggaacagga gcatcaagta gcaaaagagg
     3421 aaagagagac acctaccacc gattctatgc ccccacttct gagacaagca attgagctgt
     3481 tcgaccggca gggagccgaa cctgccttcc ttttcggcct ggaactaatc atatgtggcc
     3541 tggagaaaca gctaaagtgc gaaagcggcg ggccgaccga cgcccttgac gattttgact
     3601 tagacatgct cccagccgat gcccttgacg actttgacct tgatatgctg cctgctgacg
     3661 ctcttgacga ttttgacctt gacatgctcc ccgggtaact cgagcgcgtt aagtcgacaa
     3721 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     3781 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     3841 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     3901 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     3961 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     4021 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     4081 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     4141 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     4201 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     4261 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcgtc gactttaaga
     4321 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     4381 gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc
     4441 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     4501 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     4561 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagggcccgt
     4621 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     4681 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     4741 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     4801 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     4861 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     4921 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4981 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5041 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     5101 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     5161 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     5221 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     5281 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5341 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     5401 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     5461 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     5521 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     5581 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     5641 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     5701 tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata
     5761 tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg accagtgccg
     5821 ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg
     5881 ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc
     5941 tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg
     6001 tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg
     6061 acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc
     6121 tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc
     6181 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6241 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6301 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6361 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6421 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     6481 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6541 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6601 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6661 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6721 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6781 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6841 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6901 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6961 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7021 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     7081 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7141 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7201 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7261 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     7321 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7381 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7441 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7501 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7561 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7621 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7681 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7741 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7801 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7861 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7921 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7981 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8041 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8101 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8161 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8221 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8281 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8341 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8401 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8461 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8521 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8581 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     8641 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     8701 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     8761 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     8821 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
     8881 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     8941 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     9001 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     9061 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     9121 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     9181 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     9241 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     9301 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     9361 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     9421 acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc
     9481 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     9541 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     9601 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     9661 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
     9721 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
     9781 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     9841 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     9901 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
     9961 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
    10021 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
    10081 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
    10141 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
    10201 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
    10261 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
    10321 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
    10381 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
    10441 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
    10501 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
    10561 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
    10621 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
    10681 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
    10741 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
    10801 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
    10861 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
    10921 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
    10981 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc
    11041 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
    11101 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
    11161 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg
    11221 tta
//
