LOCUS       sequence-155360-       10512 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expressing Dox and IPTG inducible stgRNAs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10512)
  AUTHORS   Perli SD, Cui CH, Lu TK
  TITLE     Continuous genetic recording with self-targeting CRISPR-Cas in human
            cells.
  JOURNAL   Science. 2016 Aug 18. pii: aag0511.
  PUBMED    27540006
REFERENCE   2  (bases 1 to 10512)
  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: "Science. 
            2016 Aug 18. pii: aag0511."
COMMENT     Sequence Label: Construct 31 -
            1xTetO_H1p_20nt2_stgRNA_3xLacO_...
FEATURES             Location/Qualifiers
     source          1..10512
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     180..199
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     protein_bind    244..262
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     474..494
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     protein_bind    583..599
                     /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     645..664
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    676..695
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    837..856
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     promoter        898..1397
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(933..956)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1330..1349
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1415..1424
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1421..1423,1424..1426,1427..2137)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(1466..1487)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2147..2203
                     /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             2204..2575
                     /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="AKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDDV
                     TLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGRE
                     FALRDPAGNCVHFVAEEQD"
     misc_feature    2603..3191
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2656..2676)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3074..3085)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3194..3210)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3195..3211)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3716..3896
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(3922..3939)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3928..4152
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4198..4626
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4285..4304)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4495..4516
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4635..4655)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4640..4969
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4820..4955
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4882..4901
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5017..5064
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5083..5457
                     /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    5587..5708
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5624..5643)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5678..5697
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5757..5773)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5757..5773)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5770..5792)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5781..5797
                     /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(5805..5811,5812..5829,5830..5835))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5850..5871
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5988..6005)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6159..6747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6239..6258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6918..7709,7710..7778))
                     /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     7541..7560
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7779..7883)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7958..7977)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        8149..8528
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8530..8728
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8683..8703
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             8746..8926
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8973..9098
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9591..9824
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10009..10053
                     /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    10351..10468
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gcgaacgctc ctcaccatac gaaagacctt gaatcctatg cttcgaacgc tcacgtcatc
       61 aacccgctcc aaggaatcgc gggcccagtg tcactaggcg ggaacaccca gcgcgcgtgc
      121 gccctggcag gaagatggct gtgagggaca ggggagtggc gccctgcaat atttgcatgt
      181 cgctatgtgt tctgggaaat caccataaac gtgaaatgtc tttggatttg ggaatcttat
      241 aagtccctat cagtgataga gatcccagtg gctttaccaa cagtacgggt tagagctaga
      301 aatagcaagt taacctaagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
      361 gctttttttc acgaggcgga cactgattga cacggtttgc tagctgtaca aaaaagcagg
      421 ctttaaagga accaattcag tcgactggat ccggtaccaa ggtcgggcag gaagagggcc
      481 tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga gagataatta
      541 gaattaattt gactgtaaac acaaagatat tagtacaaaa aattgtgagc ggataacaat
      601 tatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta
      661 ccgtaacttg aaagtaattg tgagcgctca caattatata tcttgtggaa aggacgaaac
      721 accgagtcgc gtgtagcgaa gcagggttag agctagaaat agcaagttaa cctaaggcta
      781 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttctaga cccagcaatt
      841 gtgagcgctc acaatttaca aagttggcac tcactcgcta ttcgaactaa ttctaccggg
      901 taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca
      961 cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc
     1021 aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga
     1081 agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc
     1141 tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg
     1201 gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg
     1261 ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg
     1321 gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct
     1381 catctccggg cctttcgacc tgcatcccaa gcttgccacc atggtgagca agggcgagga
     1441 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1501 gttcagcgtg aggggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt
     1561 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgagcca
     1621 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1681 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaccta
     1741 caagacccgc gccgaggtga agttcgaggg cgacacccta gtgaaccgca tcgagctgaa
     1801 gggcgtcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacttcaa
     1861 cagccacaac atctatatca tggccgtcaa gcagaagaac ggcatcaagg tgaacttcaa
     1921 gatccgccac aacgtggagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     1981 ccccatcggc gacggccccg tgctgctgcc cgacagccac tacctgagca cccagtccgt
     2041 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tccgcaccgc
     2101 cgccgggatc actctcggca tggacgagct gtacaagggc tctggtgcaa ccaatttctc
     2161 tcttcttaaa caagccggtg atgtggagga gaaccccgga cccgccaagt tgaccagtgc
     2221 cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct
     2281 cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac
     2341 cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg
     2401 ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg
     2461 ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc
     2521 cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgataaga
     2581 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     2641 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2701 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2761 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2821 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2881 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2941 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     3001 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3061 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3121 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3181 cgcctccccg catcgatacc gtcgacctcg agacctagaa aaacatggag caatcacaag
     3241 tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga
     3301 ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt
     3361 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     3421 aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattggca
     3481 gaactacaca ccagggccag ggatcagata tccactgacc tttggatggt gctacaagct
     3541 agtaccagtt gagcaagaga aggtagaaga agccaatgaa ggagagaaca cccgcttgtt
     3601 acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtattag agtggaggtt
     3661 tgacagccgc ctagcatttc atcacatggc ccgagagctg catccggact gtactgggtc
     3721 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     3781 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     3841 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagggc
     3901 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     3961 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     4021 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     4081 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     4141 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     4201 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4261 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4321 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4381 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4441 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4501 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4561 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4621 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4681 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4741 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4801 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4861 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     4921 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     4981 agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag
     5041 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt
     5101 gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg
     5161 ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg
     5221 accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg
     5281 tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc
     5341 cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc
     5401 gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac
     5461 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5521 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5581 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5641 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5701 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     5761 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5821 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5881 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5941 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6001 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6061 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6121 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6181 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6241 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6301 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6361 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6421 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6481 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6541 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6601 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6661 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6721 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6781 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6841 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6901 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6961 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7021 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7081 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7141 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7201 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7261 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7321 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7381 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7441 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7501 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7561 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7621 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7681 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7741 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7801 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7861 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     7921 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
     7981 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     8041 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
     8101 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat
     8161 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     8221 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     8281 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     8341 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     8401 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     8461 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     8521 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     8581 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     8641 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     8701 gtgtacggtg ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta
     8761 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     8821 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     8881 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     8941 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     9001 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     9061 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     9121 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     9181 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     9241 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     9301 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     9361 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     9421 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     9481 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     9541 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     9601 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     9661 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     9721 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     9781 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     9841 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     9901 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     9961 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
    10021 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
    10081 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
    10141 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
    10201 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
    10261 gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt
    10321 ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta
    10381 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
    10441 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt
    10501 ttggttaatt aa
//
