LOCUS       sequence-472747-       11813 bp    DNA     circular SYN 03-JUN-2025
DEFINITION  Tetracycline-inducible expression of Ngn2 and Sox11 for direct 
            reprogramming of canine fibroblasts to induced-motor neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11813)
  TITLE     Protocol for direct reprogramming of canine fibroblasts to 
            induced-motor neurons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11813)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 3, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pLVX-UbC-rtTA-Ngn2_2A_Sox11-PU​RO
FEATURES             Location/Qualifiers
     source          1..11813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(115..134)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      928..1516
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1417..1436
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1670..1687
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1804..1825
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1840..1845,1846..1863,1864..1870)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1878..1894
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1883..1905
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1902..1918
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1902..1918
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     LTR             2046..2679
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    2886..3474
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2992..3003
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3401..3421
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3491..4090)
                     /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(3574..3594)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4071..4090
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(4111..4610)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4159..4178)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     4552..4575
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     regulatory      5807..5816
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5841..5894)
                     /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"
     promoter        complement(6750..7064)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     primer_bind     complement(6753..6777)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    6824..6842
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6860..6878
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6895..6913
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6931..6949
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6967..6985
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7002..7020
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7038..7056
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        7102..8313
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     8150..8169
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(8323..8940,8941..8946,8947..8982,8983..8985,8986..9021,
                     9022..9024,9025..9060,9061..9072)
                     /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
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPGN"
     misc_feature    9113..9228
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     CDS             complement(9491..9535)
                     /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    9720..9953
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    10450..10575
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             10622..11255
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    11423..11557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11447..11466)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     11501..11520
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        11605..11709
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11710..11778,11779..11813,1..757)
                     /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"
ORIGIN
        1 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
       61 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      121 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      181 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
      241 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
      301 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
      361 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
      421 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
      481 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
      541 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
      601 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
      661 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
      721 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
      781 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
      841 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
      901 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
      961 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1021 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1081 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1141 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1201 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1261 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1321 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1381 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1441 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1501 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1561 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1621 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     1681 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     1741 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     1801 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     1861 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     1921 tgattacgcc aagctctagc tagaggtcga cggtaaacgc tagcaatgtc aaggcctctc
     1981 actctctgat attcatttct ttgcaagtta taaatactga ataataagat gacatgaact
     2041 actactgcta gagattttcc acactgacta aaagggtctg agggatctct agttaccaga
     2101 gtcacacaac agacgggcac acactacttg aagcactcaa ggcaagcttt attgaggctt
     2161 aagcagtggg ttccctagtt agccagagag ctcccaggct cagatctggt ctaaccagag
     2221 agacccagta caggcaaaaa gcagctgctt atatgcagga tctgagggct cgccactccc
     2281 cagtcccgcc caggccacgc ctccctggaa agtccccagc ggaaagtccc ttgtagcaag
     2341 ctcgatatca gcagttcttg aagtactccg gatgcagctc tcgggccacg tgatgaaatg
     2401 ctaggcggct gtcaaacctc cactctaaca cttctctctc cgggtcatcc atcccatgca
     2461 ggctcacagg gtgtaacaag ctggtgttct ctcctttatt ggcctcttct accttatctg
     2521 gctcaactgg tactagcttg tagcaccatc caaaggtcag tggatatctg acccctggcc
     2581 ctggtgtgta gttctgctaa tcagggaagt agccttgtgt gtggtagatc cacagatcaa
     2641 ggatatcttg tcttcgttgg gagtgaatta gcccttccag tcccctcttt tcttttaaaa
     2701 agtggctaag atctacagct gccttgtaag tcattggtct taaaggtacc tgaggtgtga
     2761 ctggaaaacc cacctcctcc tcctcttgtg cttctagcca ggcacaatca gcattggtag
     2821 ctgctgtatt gctacttgtg attgctccat gtttttctag gtctcgactg cagaattaat
     2881 tccaggcggg gaggcggccc aaagggagat ccgactcgtc tgagggcgaa ggcgaagacg
     2941 cggaagaggc cgcagagccg gcagcaggcc gcgggaagga aggtccgctg gattgagggc
     3001 cgaagggacg tagcagaagg acgtcccgcg cagaatccag gtggcaacac aggcgagcag
     3061 ccatggaaag gacgtcagct tccccgacaa caccacggaa ttgtcagtgc ccaacagccg
     3121 agcccctgtc cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg caatagggag
     3181 ggggaaagcg aaagtcccgg aaaggagctg acaggtggtg gcaatgcccc aaccagtggg
     3241 ggttgcgtca gcaaacacag tgcacaccac gccacgttgc ctgacaacgg gccacaactc
     3301 ctcataaaga gacagcaacc aggatttata caaggaggag aaaatgaaag ccatacggga
     3361 agcaatagca tgatacaaag gcattaaagc agcgtatcca catagcgtaa aaggagcaac
     3421 atagttaaga ataccagtca atctttcaca aattttgtaa tccagaggtt gattgttcca
     3481 gacgcgcccg tcaggcaccg ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca
     3541 cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta gaaggggagg ttgcggggcg
     3601 cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc cgcctccact ccggggagca
     3661 cgacggcgct gcccagaccc ttgccctggt ggtcgggcga gacgccgacg gtggccagga
     3721 accacgcggg ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt tgctgcgcgg
     3781 ccagccggga accgctcaac tcggccatgc gcgggccgat ctcggcgaac accgcccccg
     3841 cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc gccgtcgtcc gcgacccaca
     3901 ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct
     3961 cgatgtggcg gtccggatcg acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg
     4021 cgagggtgcg tacggccctg gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt
     4081 actcggtcat ggtaagcttg ggctgcaggt cgaaaggccc ggagatgagg aagaggagaa
     4141 cagcgcggca gacgtgcgct tttgaagcgt gcagaatgcc gggcctccgg aggaccttcg
     4201 ggcgcccgcc ccgcccctga gcccgcccct gagcccgccc ccggacccac cccttcccag
     4261 cctctgagcc cagaaagcga aggagcaaag ctgctattgg ccgctgcccc aaaggcctac
     4321 ccgcttccat tgctcagcgg tgctgtccat ctgcacgaga ctagtgagac gtgctacttc
     4381 catttgtcac gtcctgcacg acgcgagctg cggggcgggg gggaacttcc tgactagggg
     4441 aggagtagaa ggtggcgcga aggggccacc aaagaacgga gccggttggc gcctaccggt
     4501 ggatgtggaa tgtgtgcgag gccagaggcc acttgtgtag cgccaagtgc ccagcggggc
     4561 tgctaaagcg catgctccag actgccttgg gaaaagcgcc tcccctaccc ggtagaattc
     4621 actcaatacg tgaacaccag gtcggagaag ttcgcctcca gccagtcccc cgcgatcatc
     4681 tcgctcagct ccggcgtgca gtagtcgggg aactcgaagt gggaacccag gctgccctcg
     4741 ctgaaggaat ccaggtcctt atccaccagc gacagggaca ggttccccgc cgctcccgcg
     4801 cccagtggct gctcgcaggc gctgtgcgcg ccctgggaga aattcaagct caggtcgaac
     4861 atgaggtcgt ctgcatcctc ggcgccgctg ccgctgctgc tgccgctgga tgaggaggtg
     4921 gacacgcagc gggaggacgc gggcgacagc gcgggaggcg cgggcggagg ctgctgcttg
     4981 gtgatgttct tgaagctgta gtagagccgg ccgcccgcgc gcacttcgtc gtacaggctc
     5041 gcgccctcgg gggactcggc ggcactgctg agcgtggggc tggcggggac cttggccacg
     5101 ctgtagcgcc tcagcagctg agggggttgc tgctgcgtcg gcggcggcag caggtgcgag
     5161 tgcgcgggct cgtcgtcgtc gtcgtcagcg tccggcttgg gccgtagctg cagctcgtca
     5221 tcttcgtcgt cgtcttcatc gtcgtcgtcc aggaagacgc acttggctgc cttgcccgcg
     5281 gcgcagtccc ccggctgcgc cgccttgccc gcgcccgcct tgcccgcagg cgccttgagc
     5341 ttggcgcact tcttgccggg gcccttggct gccttggcgc ccgccgcgct cttgtcgggg
     5401 ctctggcccg cgctgggctt ggccgctggg tccgtcttgg gctttttgcg cggccggtac
     5461 ttgtagtcgg gataatcagc catgtgcttg aggcgcaggc gctccgcctc cctgatgaac
     5521 gggatcttct cgctgtcctt cagcatcttc cagcgcttgc ccagcctctt ggagatctcg
     5581 gcgttgtgca tgtcgggcga ctgctccatg atcttcctgc gctcgatctt ggaccacacc
     5641 atgaaggcgt tcatgggccg tttgatgtgg cccgacgccg tcttgcacca gtccgggtcg
     5701 ctctcgtcca gggccaccgg gctgcacgcc atgaactcgc cctcctcggt gtccagcgcg
     5761 tcccggggca ggttgctctc ggcttccgag ctctcggcct gctgcaccat ggtggcggat
     5821 ctgagtccgg tagcgctagc gggtccagga ttctcttcga cgtctccgca ggtcagaaga
     5881 cttcctctcc cctcgtacat atggatacaa tccctggcta tggggaggtg aggtgcatag
     5941 cggtgcttgt cgggaggtgg gggctgccaa tagtccatgt ctgacccggc cgggctggcg
     6001 ggcgataaag tgcagctgta gggggaggtg gaattggagg acacggagga ggacggcgcg
     6061 gggctgttgg tgcaactcca cgtggaggcg ggcgaggggc tgtctccgct gctgctcagg
     6121 gcggcgctgg cgcctcccgg gctcagcaac actgcctcgg agaagagcgc ccccggcagg
     6181 cccccgccgc cgcccccgca gtgatccgcc aggcgcaggg tctcggtgag tgcccagatg
     6241 tagttgtggg cgaagcgcag ggtctcgatc ttggtgagct tggcgtcctc ggggaacgtg
     6301 gggagcacct cgcgcagcgc gtccagtgcc gcgttgaggt tgtgcatgcg gtttcgctcg
     6361 cggttgttgg ccttcagtct acgggtcttc ttgatgcgct gcaccgtctc ggccgtcttg
     6421 gcgcctcggg agacggcccg cgcccgggaa gggcgccgtt tgcaatcgtg taccagaccc
     6481 agcagccgtg cgggccggca gccctccgca cccgcagcca cgccgccccg cgccccctgc
     6541 ccggcctcag ccccgcgctg ccgacgcgcc ccgcctgacg cgcccggctc ctcctcctct
     6601 tcttcgtcgg cgctggatga cagcggggtc agggccgcca aggcggggga ggccgatccg
     6661 agcagcacta acacgtcctc ttcctccttc aactccaagg tctcggattt gacgaacatg
     6721 gtggcggcgc ggccgcggat ccttctccag gcgatctgac ggttcactaa acgagctctg
     6781 cttatatagg cctcccaccg tacacgccta cctcgacata cgttctctat cactgatagg
     6841 gagtaaactc gacatacgtt ctctatcact gatagggata aactcgacat acgttctcta
     6901 tcactgatag ggagtaaact cgacatacgt tctctatcac tgatagggag taaactcgac
     6961 atacgttctc tatcactgat agggagtaaa ctcgacatcg ttctctatca ctgataggga
     7021 gtaaactcga catacgttct ctatcactga tagggagtaa actcgagtga agacgaaagg
     7081 gcctcatcga tgcgccgatc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc
     7141 ccctcctcac ggcgagcgct gccacgtcag acgaagggcg cagcgagcgt cctgatcctt
     7201 ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt
     7261 atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc
     7321 cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc
     7381 cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc
     7441 cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg
     7501 tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac
     7561 ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg
     7621 aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga
     7681 cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag
     7741 aacccaaggt cttgaggcct tcgctaatgc gggaaagctc ttattcgggt gagatgggct
     7801 ggggcaccat ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc
     7861 gtctgttgcg ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg
     7921 agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg
     7981 ggccacctgc cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc
     8041 ctagggtagg ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg
     8101 aggcgtcagt ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg
     8161 ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt
     8221 gaaatgtaat catttgggtc aatatgtaat tttcagtgtt agactagtaa attgtccgct
     8281 aaattctggc cgtttttggc ttttttgtta gacggatcca ccatgtctag actggacaag
     8341 agcaaagtca taaacggcgc tctggaatta ctcaatggag tcggtatcga aggcctgacg
     8401 acaaggaaac tcgctcaaaa gctgggagtt gagcagccta ccctgtactg gcacgtgaag
     8461 aacaagcggg ccctgctcga tgccctgcca atcgagatgc tggacaggca tcatacccac
     8521 ttctgccccc tggaaggcga gtcatggcaa gactttctgc ggaacaacgc caagtcattc
     8581 cgctgtgctc tcctctcaca tcgcgacggg gctaaagtgc atctcggcac ccgcccaaca
     8641 gagaaacagt acgaaaccct ggaaaatcag ctcgcgttcc tgtgtcagca aggcttctcc
     8701 ctggagaacg cactgtacgc tctgtccgcc gtgggccact ttacactggg ctgcgtattg
     8761 gaggaacagg agcatcaagt agcaaaagag gaaagagaga cacctaccac cgattctatg
     8821 cccccacttc tgagacaagc aattgagctg ttcgaccggc agggagccga acctgccttc
     8881 cttttcggcc tggaactaat catatgtggc ctggagaaac agctaaagtg cgaaagcggc
     8941 gggccggccg acgcccttga cgattttgac ttagacatgc tcccagccga tgcccttgac
     9001 gactttgacc ttgatatgct gcctgctgac gctcttgacg attttgacct tgacatgctc
     9061 ccgggtaact aatcgataaa ctggatctct gctgtccctg taataaaccc gaaaattttg
     9121 aatttttgta atttgttttt gtaattcttt agtttgtatg tctgttgcta ttatgtctac
     9181 tattctttcc cctgcactgt accccccaat cccccctttt cttttaaagg cgataccgtc
     9241 gagatccgtt cactaatcga atggatctgt ctctgtctct ctctccacct tcttcttcta
     9301 ttccttcggg cctgtcgggt cccctcgggg ttgggaggtg ggtctgaaac gataatggtg
     9361 aatatccctg cctaactcta ttcactatag aaagtacagc aaaaactatt cttaaaccta
     9421 ccaagcctcc tactatcatt atgaataatt ttatatacca cagccaattt gttatgttaa
     9481 accaattcca caaacttgcc catttatcta attccaataa ttcttgttca ttcttttctt
     9541 gctggttttg cgattcttca attaaggagt gtattaagct tgtgtaattg ttaatttctc
     9601 tgtcccactc catccaggtc gtgtgattcc aaatctgttc cagagattta ttactccaac
     9661 tagcattcca aggcacagca gtggtgcaaa tgagttttcc agagcaaccc caaatcccca
     9721 ggagctgttg atcctttagg tatctttcca cagccaggat tcttgcctgg agctgcttga
     9781 tgccccagac tgtgagttgc aacagatgct gttgcgcctc aatagccctc agcaaattgt
     9841 tctgctgctg cactatacca gacaataatt gtctggcctg taccgtcagc gtcattgacg
     9901 ctgcgcccat agtgcttcct gctgctccca agaacccaag gaacaaagct cctattccca
     9961 ctgctctttt ttctctctgc accactcttc tctttgcctt ggtgggtgct actcctaatg
    10021 gttcaatttt tactacttta tatttatata attcacttct ccaattgtcc ctcatatctc
    10081 ctcctccagg tctgaagatc agcggccggc cgcttgctgt gcggtggtct tacttttgtt
    10141 ttgctcttcc tctatcttgt ctaaagcttc cttggtgtct tttatctcta tcctttgatg
    10201 cacacaatag agggttgcta ctgtattata taatgatcta agttcttctg atcctgtctg
    10261 aagggatggt tgtagctgtc ccagtatttg tctacagcct tctgatgttt ctaacaggcc
    10321 aggattaact gcgaatcgtt ctagctccct gcttgcccat actatatgtt ttaatttata
    10381 ttttttcttt ccccctggcc ttaaccgaat tttttcccat cgcgatctaa ttctcccccg
    10441 cttaatactg acgctctcgc acccatctct ctccttctag cctccgctag tcaaaatttt
    10501 tggcgtactc accagtcgcc gcccctcgcc tcttgccgtg cgcgcttcag caagccgagt
    10561 cctgcgtcga gagagctcct ctggtttccc tttcgctttc aagtccctgt tcgggcgcca
    10621 ctgctagaga ttttccacac tgactaaaag ggtctgaggg atctctagtt accagagtca
    10681 cacaacagac gggcacacac tacttgaagc actcaaggca agctttattg aggcttaagc
    10741 agtgggttcc ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac
    10801 ccagtacagg caaaaagcag ctgcttatat gcaggatctg agggctcgcc actccccagt
    10861 cccgcccagg ccacgcctcc ctggaaagtc cccagcggaa agtcccttgt agcaagctcg
    10921 atatcagcag ttcttgaagt actccggatg cagctctcgg gccacgtgat gaaatgctag
    10981 gcggctgtca aacctccact ctaacacttc tctctccggg tcatccatcc catgcaggct
    11041 cacagggtgt aacaagctgg tgttctctcc tttattggcc tcttctacct tatctggctc
    11101 aactggtact agcttgtagc accatccaaa ggtcagtgga tatctgaccc ctggccctgg
    11161 tgtgtagttc tgctaatcag ggaagtagcc ttgtgtgtgg tagatccaca gatcaaggat
    11221 atcttgtctt ctttgggagt gaattagccc ttccaactac taagtttgta gtacatattt
    11281 aacaaataca atttctttaa aatgaaaata attcagagga atcacaggtt tagagtaaat
    11341 gaaaccacag gtaattggca gtggtaatag ggtatggggt gggaagtttg ggatgatttt
    11401 ggttagcttg agttatccag ttgatccaga catgataaga tacattgatg agtttggaca
    11461 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
    11521 tttatttgta accattataa gctgcaataa acaagttgat ctcccgatcc gtcgacgtca
    11581 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
    11641 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
    11701 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
    11761 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tga
//
