LOCUS       sequence-335358-       10211 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encoding the TRE promoter based eccDNA biosensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10211)
  AUTHORS   Moller HD, Lin L, Xiang X, Petersen TS, Huang J, Yang L, Kjeldsen E,
            Jensen UB, Zhang X, Liu X, Xu X, Wang J, Yang H, Church GM, Bolund 
            L, Regenberg B, Luo Y
  TITLE     CRISPR-C: circularization of genes and chromosome by CRISPR in human
            cells.
  JOURNAL   Nucleic Acids Res. 2018 Dec 14;46(22):e131. doi: 10.1093/nar/gky767.
  PUBMED    30551175
REFERENCE   2  (bases 1 to 10211)
  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: "Nucleic 
            Acids Res."; date: "2018-12-14"; volume: "46(22):e131. doi"; pages: 
            " 10.1093/nar/gky767"
COMMENT     Sequence Label: pPB-TRE-ECC-Biosensor
FEATURES             Location/Qualifiers
     source          1..10211
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..19
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    37..55
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    73..91
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    109..127
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    145..163
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    181..199
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    217..235
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     280..304
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          428..656
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     promoter        complement(657..934)
                     /label=chicken beta-actin promoter
     enhancer        936..1221
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     polyA_signal    1298..1522
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1511..1528
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     primer_bind     1541..1557
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1541..1557
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             complement(join(1582..2295,2296..2298,2299..2301))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1585..2295,2296..2298,2299..2301))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1627..1648)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1976..1995
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2235..2256
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2298..2307
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2372..2381
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2378..3088
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2523..2541)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2791..2810
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3110..3129)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3154..3202
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        3433..3644
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3657..3925
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3959..4702
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             4703..4756
                     /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             4757..5356
                     /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(4757..4776)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5253..5273
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5389..5510
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     5480..5499
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   6273..6307
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6798..6814)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6798..6814)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6811..6833)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6822..6838
                     /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(6846..6852,6853..6870,6871..6876))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6891..6912
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7029..7046)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7200..7788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7280..7299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7959..8750,8751..8819))
                     /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     8582..8601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8820..8924)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8950..9405)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9087..9108)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9299..9318
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9532..9554
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9546..9563
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9547..9563
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   9693..9755
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        join(10204..10211,1..371)
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
ORIGIN
        1 tccctatcag tgatagagaa cgtatgaaga gtttactccc tatcagtgat agagaacgta
       61 tgcagacttt actccctatc agtgatagag aacgtataag gagtttactc cctatcagtg
      121 atagagaacg tatgaccagt ttactcccta tcagtgatag agaacgtatc tacagtttac
      181 tccctatcag tgatagagaa cgtatatcca gtttactccc tatcagtgat agagaacgta
      241 taagctttag gcgtgtacgg tgggcgccta taaaagcaga gctcgtttag tgaaccgtca
      301 gatcgcctgg agcaattcca caacactttt gtcttatacc aactttccgt accacttcct
      361 accctcgtaa aggtctagag ctagcgaatt cggggcacag agccccagag accggcaccg
      421 gtccaacctg aaaaaaagtg atttcaggca ggtgctccag gtaattaaac attaataccc
      481 caccaaccaa ccatccctta aacccttacc tcttgctcag ctaattacag cccggaggag
      541 aagggccgtc ccgcccgctc acctgtggga gtaacgcggt cagtcagagc cggggcgggc
      601 ggcgcgaggc ggcggcggag cggggcacgg ggcgaaggca gcgtcgcagc gactcccgcc
      661 cgccgcgcgc ttcgcttttt atagggccgc cgccgccgcc gcctcgccat aaaaggaaac
      721 tttcggagcg cgccgctctg attggctgcc gccgcacctc tccgcctcgc cccgccccgc
      781 cccccccccc cccccccccc ccccggcgcg cgcccccccc ccccccccgc ccccatcgct
      841 gcacaaaata attaaaaaat aaataaatac aaaattgggg gtggggaggg gggggagatg
      901 gggagagtga agcagaacgt ggggctcacc tcgaccatgg taatagcgat gactaatacg
      961 tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcaca atgccaggcg
     1021 ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt
     1081 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
     1141 attggcgtta ctattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
     1201 tttaccgtaa gttatgtaac gggtacctct agagccattt gtctgcaata aagcaatagc
     1261 atcacaaatt tcacaaataa agcatttttt tcactgccca tagagcccac cgcatcccca
     1321 gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc
     1381 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
     1441 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
     1501 ggctggcaac tagaaggcac agtcgaggct gatcggccgc tctagaacta gtggatcccc
     1561 cgggctgcat ccaacctcga gttacttgta cagctcgtcc atgccgagag tgatcccggc
     1621 ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag
     1681 ggcggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg
     1741 ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt tgtggcggat
     1801 cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga cgttgtggct
     1861 gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc
     1921 cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt
     1981 gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc
     2041 ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc actgcacgcc
     2101 gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat
     2161 gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa
     2221 cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag
     2281 ctcctcgccc ttgctcacca tggtggcggt gcccgggaga cgccaaccgc aagccgggac
     2341 gcgttcaagc ttcgcctgga gacgatccgc cgccaccatg gtgagcaagg gcgaggagga
     2401 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
     2461 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     2521 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
     2581 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
     2641 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     2701 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     2761 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     2821 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
     2881 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
     2941 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
     3001 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     3061 cggcggcatg gacgagctgt acaagtaagt ttaaacgggg gaggctaact gaaacacgga
     3121 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     3181 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     3241 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     3301 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     3361 gccatagcag atctgcgaaa tcggatccag ctgaatctaa gtggatctgc gatcgctccg
     3421 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
     3481 tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
     3541 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
     3601 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag cttcgagggg
     3661 ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca cgccggttga
     3721 gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa
     3781 gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc ctacctagac
     3841 tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc tttgtttcgt
     3901 tttctgttct gcgccgttac agatctaagc tgtgaccggc gcctactcta gagccgccat
     3961 gtctagactg gacaagagca aagtcataaa ctctgctctg gaattactca atggagtcgg
     4021 tatcgaaggc ctgacgacaa ggaaactcgc tcaaaagctg ggagttgagc agcctaccct
     4081 gtactggcac gtgaagaaca agcgggccct gctcgatgcc ctgccaatcg agatgctgga
     4141 caggcatcat acccactcct gccccctgga aggcgagtca tggcaagact ttctgcggaa
     4201 caacgccaag tcataccgct gtgctctcct ctcacatcgc gacggggcta aagtgcatct
     4261 cggcacccgc ccaacagaga aacagtacga aaccctggaa aatcagctcg cgttcctgtg
     4321 tcagcaaggc ttctccctgg agaacgcact gtacgctctg tccgccgtgg gccactttac
     4381 actgggctgc gtattggagg aacaggagca tcaagtagca aaagaggaaa gagagacacc
     4441 taccaccgat tctatgcccc cacttctgaa acaagcaatt gagctgttcg accggcaggg
     4501 agccgaacct gccttccttt tcggcctgga actaatcata tgtggcctgg agaaacagct
     4561 aaagtgcgaa agcggcgggc cgaccgacgc ccttgacgat tttgacttag acatgctccc
     4621 agccgatgcc cttgacgact ttgaccttga tatgctgcct gctgacgctc ttgacgattt
     4681 tgaccttgac atgctccccg gggagggcag aggaagtctt ctaacatgcg gtgacgtgga
     4741 ggagaatccc ggccctatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     4801 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     4861 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     4921 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     4981 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     5041 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     5101 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     5161 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     5221 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     5281 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     5341 caagcccggt gcctgaaatc aacctctgga ttacaaaatt tgtgggtaaa cttgtttatt
     5401 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5461 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     5521 aattgactca aatgatgtca attagtctat cagaagctca tctggtctcc cttccggggg
     5581 acaagacatc cctgtttaat atttaaacag cagtgttccc aaactgggtt cttatatccc
     5641 ttgctctggt caaccaggtt gcagggtttc ctgtcctcac aggaacgaag tccctaaaga
     5701 aacagtggca gccaggttta gccccggaat tgactggatt ccttttttag ggcccattgg
     5761 tatggctttt tccccgtatc cccccaggtg tctgcaggct caaagagcag cgagaagcgt
     5821 tcagaggaaa gcgatcccgt gccaccttcc ccgtgcccgg gctgtccccg cacgctgccg
     5881 gctcggggat gcggggggag cgccggaccg gagcggagcc ccgggcggct cgctgctgcc
     5941 ccctagcggg ggagggacgt aattacatcc ctgggggctt tggggggggg ctgtccctga
     6001 tatctataac aagaaaatat atatataata agttatcacg taagtagaac atgaaataac
     6061 aatataatta tcgtatgagt taaatcttaa aagtcacgta aaagataatc atgcgtcatt
     6121 ttgactcacg cggtcgttat agttcaaaat cagtgacact taccgcattg acaagcacgc
     6181 ctcacgggag ctccaagcgg cgactgagat gtcctaaatg cacagcgacg gattcgcgct
     6241 atttagaaag agagagcaat atttcaagaa tgcatgcgtc aattttacgc agactatctt
     6301 tctagggtta atctagctgc atcaggatca tatcgtcggg tcttttttcc ggctcagtca
     6361 tcgcccaagc tggcgctatc tgggcatcgg ggaggaagaa gcccgtgcct tttcccgcga
     6421 ggttgaagcg gcatggaaag agtttgccga ggatgactgc tgctgcattg acgttgagcg
     6481 aaaacgcacg tttaccatga tgattcggga aggtgtggcc atgcacgcct ttaacggtga
     6541 actgttcgtt caggccacct gggataccag ttcgtcgcgg cttttccgga cacagttccg
     6601 gatggtcagc ccgaagcgca tcagcaaccc gaacaatacc ggcgacagcc ggaactgccg
     6661 tgccggtgtg cagattaatg acagcggtgc ggcgctggga tattacgtca gcgaggacgg
     6721 gtatcctggc tggatgccgc agaaatggac atggataccc cgtgagttac ccggcgggcg
     6781 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     6841 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6901 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6961 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     7021 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     7081 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     7141 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     7201 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     7261 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     7321 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7381 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7441 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7501 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7561 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7621 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7681 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7741 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7801 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7861 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7921 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7981 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     8041 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     8101 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     8161 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     8221 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     8281 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     8341 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     8401 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     8461 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8521 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8581 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8641 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8701 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8761 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8821 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8881 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8941 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     9001 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     9061 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     9121 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     9181 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     9241 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     9301 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     9361 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca
     9421 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     9481 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     9541 gacgttgtaa aacgacggcc agtgagcgcg cctcgttcat tcacgttttt gaacccgtgg
     9601 aggacgggca gactcgcggt gcaaatgtgt tttacagcgt gatggagcag atgaagatgc
     9661 tcgacacgct gcagaacacg cagctagatt aaccctagaa agataatcat attgtgacgt
     9721 acgttaaaga taatcatgtg taaaattgac gcatgtgttt tatcggtctg tatatcgagg
     9781 tttatttatt aatttgaata gatattaagt tttattatat ttacacttac atactaataa
     9841 taaattcaac aaacaattta tttatgttta tttatttatt aaaaaaaaca aaaactcaaa
     9901 atttcttcta taaagtaaca aaacttttat gagggacagc ccccccccaa agcccccagg
     9961 gatgtaatta cgtccctccc ccgctagggg gcagcagcga gccgcccggg gctccgctcc
    10021 ggtccggcgc tccccccgca tccccgagcc ggcagcgtgc ggggacagcc cgggcacggg
    10081 gaaggtggca cgggatcgct ttcctctgaa cgcttctcgc tgctctttga gcctgcagac
    10141 acctgggggg atacggggaa aaggcctcca cggccactag tctttcgtct tcaagaattc
    10201 ctcgagttta c
//
