LOCUS       sequence-389716-       13386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid for CRISPR/Cas9-mediated insertion of INSPECT into 
            human IL2 locus (exon 3). Encodes for intronic IRES-driven 
            SigP:NLuc. Plasmid contains FRT-site flanked PuroR-TK cassette..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13386)
  AUTHORS   Truong DJ, Armbrust N, Geilenkeuser J, Lederer EM, Santl TH, Beyer 
            M, Ittermann S, Steinmassl E, Dyka M, Raffl G, Phlairaharn T, 
            Greisle T, Zivanic M, Grosch M, Drukker M, Westmeyer GG
  TITLE     Intron-encoded cistronic transcripts for minimally invasive 
            monitoring of coding and non-coding RNAs.
  JOURNAL   Nat Cell Biol. 2022 Nov;24(11):1666-1676. doi: 
            10.1038/s41556-022-00998-6. Epub 2022 Nov 7.
  PUBMED    36344775
REFERENCE   2  (bases 1 to 13386)
  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"; doi: 
            "10.1038/s41556-022-00998-6"; journalName: "Nat Cell Biol"; date: 
            "2022-11"; volume: "24"; issue: "11"; pages: "1666-1676"
COMMENT     Sequence Label: pINSPECT:IL2_SigP:NLuc
FEATURES             Location/Qualifiers
     source          1..13386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2153..2186
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2257..2290
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     polyA_signal    2889..3010
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2926..2945)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2980..2999
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3042..3061
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    3060..3115
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     protein_bind    complement(3222..3255)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3326..3359)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             complement(3408..3464)
                     /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             complement(4599..4655)
                     /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"
     primer_bind     5242..5261
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(5288..6470)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5297..6239
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(5320..5340)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    join(6546..7119,7120..7122,7123..7131)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6713..6730)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6940..6959
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(11198..11218)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(11198..11216)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11237..11253)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11237..11253)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11250..11272)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11261..11277
                     /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(11285..11291,11292..11309,11310..11315))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11330..11351
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11468..11485)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11639..12227)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11719..11738)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12398..13189,13190..13258))
                     /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     13021..13040
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13259..13363)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcgt
      661 ctcggacaca gatgcaatcc cattgactca aattctatta attcaagaga aatctgcttc
      721 taactaccct tctgaaagat gtaaaggaga cagcttacag atgttactct agtttaatca
      781 gagccacata atgcaactcc agcaacataa agatactaga tgctgttttc tgaagaaaat
      841 ttctccacat tgttcatgcc aaaaacttaa acccgaattt gtagaatttg tagtggtgaa
      901 ttgaaagcgc aatagatgga catatcaggg gattggtatt gtcttgacct acctttccca
      961 ctaaagagtg ttagaaagat gagattatgt gcataattta gggggtggta gaattcatgg
     1021 aaatctaagt ttgaaaccaa aagtaatgat aaactctatt catttgttca tttaaccctc
     1081 attgcacatt tacaaaagat tttagaaact aataaaaata tttgattcca aggatgctat
     1141 gttaatgcta taatgagaaa gaaatgaaat ctaattctgg ctctacctac ttatgtggtc
     1201 aaattctgag atttagtgtg cttatttata aagtggagat gatacttcac tgcctacttc
     1261 aaaagatgac tgtgagaagt aaatgggcct attttggaga aaattctttt aaattgtaat
     1321 ataccataga aatatgaaat attatatata atatagaatc aagaggcctg tccaaaagtc
     1381 ctcccaaagt attataatct tttatttcac tgggacaaac atttttaaaa tgcatcttaa
     1441 tgtagtgatt gtagaaaagt aaaaatttaa gacatattta aaaatgtgtc ttgctcaagg
     1501 ctatattgag agccactact acatgattat tgttacctag tgtaaaatgt tgggattgtg
     1561 atagatggca tccaagagtt ccttctctct caacattctg tgattcttaa ctcttagact
     1621 atcaaatatt ataatcatag aatgtgattt ttatgcttcc acattctaac tcatctggtt
     1681 ctaatgattt tctatgcaga ttggaaaagt aatcagccta catctgtaat aggcatttag
     1741 atgcagaaag tctaacattt tgcaaagcca aattaagcta aaaccagtga gtcaactatc
     1801 acttaacgct agtcataggt acttgagccc tagtttttcc agttttataa tgtaaactct
     1861 actggtccat ctttacagtg acattgagaa cagagagaat ggtaaaaact acatactgct
     1921 actccaaata aaataaattg gaaattaatt tctgattctg acctctatgt aaactgagct
     1981 gatgataatt attattctag gccacagaac tgaaacatct tcagtgtcta gaagaagaac
     2041 tcaaacctct ggaggtaagt atccttttta cagcacaact taatgagaca gatagaaact
     2101 ggtcttgtag aaacagagta gattttatcg aattgcaacc tcttcgaggg acataacttc
     2161 gtataatgta tgctatacga agttatgggt cgatgctgag atctggacta gagggtcgat
     2221 ggtgatgctt ggatcaaaat aggaagaatc ccaatgataa cttcgtataa agtatcctat
     2281 acgaagttat actagaattg cgtaccgcca ccatgtcgcc aaggccacct cccctgtgag
     2341 ctagactgga cagccaatga cgggtaagag agtgacattt ctcactaacc taagacagga
     2401 gggccgtcaa agctactgcc taatccaatg acgggtaata gtgacaagaa atgtatcact
     2461 ccaacctaag acaggcgcag cctccgaggg atgtgtcttt tgttttttat aattaaaaag
     2521 ggtgacacgt gaggccacct cccctgtgag ctagactgga cagccaatga cgggtaagag
     2581 agtgacattt ctcactaacc taagacagga gggccgtcaa agctactgcc taatccaatg
     2641 acgggtaata gtgacaagaa atgtatcact ccaacctaag acaggcgcag cctccgaggg
     2701 atgtgtcttt tgttttttat aattaaaaag ggtgaagccg acacacaaaa aaccaacaca
     2761 cagatctcta gagctctgga tcttttattg cccgggctac cacatttgta gaggttttac
     2821 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     2881 ttgttgttaa cttgtttatt gcagcttata atggttacaa atatagcaat agcatcacaa
     2941 atttcacaaa tatagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3001 atgtatctta tcatgtctgt atactttaaa tgatttgccc tcccatatgt ccttccgagt
     3061 gagagacaca aaaaattcca acacactatt gcaatgaaaa tacatttcct ttattagcca
     3121 gaagtcagat gccagcacaa caaccagcac gttgcccagg agctgtagga aaaagaagaa
     3181 ggcatgaaca tagttagcag agttacgtag cgatcgctag tataacttcg tatagcatac
     3241 attatacgaa gttatgggtc gatggtgaca atagcgaaaa agaaagaaca atcaagggtc
     3301 cccaaactca cctaggtcct ttgatataac ttcgtatagg atactttata cgaagttatg
     3361 gcatctcctt cggaagttcc tattctattt gaagtatagg aacttccagg tccaggattt
     3421 tcctcgacgt cccctgcctg cttgagcagt gaaaagttag ttgccccgga tccgttggct
     3481 tcgcccatct ctctagcgaa ggttctggcg aggtcacaga tggtggggat gctgccaggg
     3541 gttgtcacat gggtctgcac catgccgctt gtcagttgca gcagggcatc tctgcatccg
     3601 gctgggctct gatcgtagtc caggatgaac acatgcatgg gccgcagtct ttttgccagc
     3661 acgtcgagag cccaggcgaa cacgttatac aggtcgccgt taggggccag cagttcaggg
     3721 gctctgaaca gtgtgaacag ggtgtcgccg atgtgaggtc tgggtccagc gttagactga
     3781 ggctcagcgc cttgtggagg aactgctgtg ccagagagct gtccccagtc ctctctccag
     3841 gaaccgccgc cttgaagata gcgcacggtg ttagccagca ggccgtacac tcttctgatg
     3901 gcggccagca tagccagatc cagtctctcg ccaggcctct gtctcttagc cagtctgtcg
     3961 atgtgtctgt cctctggcag ggctccaagc acgatgtttg tgccgggcag tgtaggagga
     4021 atcagagcca cgaaagccag cacggcctga ggtgtcatgc tgcccatcag ataccgagcg
     4081 gcagggtaac acagcagggc gaagatgggg tgtctgtcga agatcagggt cagagcaggt
     4141 ggtggagcgt gagaagatcc agcctcgccg ccaatatgtg gagccagcac agcgtctgtc
     4201 acggcgtaag gcatgcccat tgtgatctgg gcagatgtca tgaccacagc agcgtcgcct
     4261 gcgctaattt cgccctggtc cagtctgtgc tgtgtggtgt agatgttggc gattgtctca
     4321 gaggctccca ggacctgcca gtaggtcata ggctcgggca cgtacacgat gtcatctctg
     4381 ctgcccagag ccaccagcag ctgtgttgtt gtggtcttgc ccatgccatg agggccgtcg
     4441 atatacactc tcagcagtgt gggcatcttc tgttccagcc gcacttctgt agcctcttgc
     4501 tgccttcttg gcctgagggc tgttcttctg ttgctgtggc ctctgcttct agcggcttga
     4561 tcgaaggcgc tggcgtgctg gtgacaagga taagaagcag gaccggggtt ctcttccacg
     4621 tcgccagcct gtttcagcag gctgaagttt gtagcgccgg atccggcgcc aggctttctg
     4681 gtcatgcacc aggttctagg gccctcaggc acttccacgt cggcggtcac ggtgaagccc
     4741 agtctctcgt agaagggcag gtttctgggg gcgcttgttt ccaggaaggc gggcacgcca
     4801 gccctttcag cagcttccac cccaggcagc accacagcag atcccagtcc cttgccctgg
     4861 tggtcaggtg acacgcccac ggtggccaga aaccaggcag gctcttttgg tctgtggggg
     4921 gccagcaggc cttccatctg ctgctgggca gccagtctag agccgctcag ctcggccatt
     4981 ctaggtccga tctcggcgaa cacagcgccg gcttccacag actcaggggt tgtccacaca
     5041 gccacagcgg cgccatcatc ggccacccac actttgccga tgtccaggcc cactctggtc
     5101 agaaacagtt cctgcagctc ggtcactctc tcgatgtgcc ggtcggggtc cacggtgtgt
     5161 cttgtggcag ggtaatcggc gaaggcagcg gccagtgtcc gcacagctct tggcacatcg
     5221 tccctggtgg ccagccgcac tgtgggcttg tactcggtca tggtggcgcg ccttttaggg
     5281 gtagttttca cgacacctga aatggaagaa aaaaactttg aaccactgtc tgaggcttga
     5341 gaatgaacca agatccaaac tcaaaaaggg caaattccaa ggagaattac atcaagtgcc
     5401 aagctggcct aacttcagtc tccacccact cagtgtgggg aaactccatc gcataaaacc
     5461 cctcccccca acctaaagac gacgtactcc aaaagctcga gaactaatcg aggtgcctgg
     5521 acggcgcccg gtactccgtg gagtcacatg aagcgacggc tgaggacgga aaggcccttt
     5581 tcctttgtgt gggtgactca cccgcccgct ctcccgagcg ccgcgtcctc cattttgagc
     5641 tccctgcagc agggccggga agcggccatc tttccgctca cgcaactggt gccgaccggg
     5701 ccagccttgc cgcccagggc ggggcgatac acggcggcgc gaggccaggc accagagcag
     5761 gccggccagc ttgagactac ccccgtccga ttctcggtgg ccgcgctcgc aggccccgcc
     5821 tcgccgaaca tgtgcgctgg gacgcacggg ccccgtcgcc gcccgcggcc ccaaaaaccg
     5881 aaataccagt gtgcagatct tggcccgcat ttacaagact atcttgccag aaaaaaagcg
     5941 tcgcagcagg tcatcaaaaa ttttaaatgg ctagagactt atcgaaagca gcgagacagg
     6001 cgcgaaggtg ccaccagatt cgcacgcggc ggccccagcg cccaggccag gcctcaactc
     6061 aagcacgagg cgaaggggct ccttaagcgc aaggcctcga actctcccac ccacttccaa
     6121 cccgaagctc gggatcaaga atcacgtact gcagccaggg gcgtggaagt aattcaaggc
     6181 acgcaagggc cataacccgt aaagaggcca ggcccgcggg aaccacacac ggcacttacc
     6241 tgtgttctgg cggcaaaccc gttgcgaaaa agaacgttca cggcgactac tgcacttata
     6301 tacggttctc ccccaccctc gggaaaaagg cggagccagt acacgacatc actttcccag
     6361 tttaccccgc gccaccttct ctaggcaccg gttcaattgc cgacccctcc ccccaacttc
     6421 tcggggactg tgggcgatgt gcgctctgcc cactgacggg caccggagcc aattcgaagt
     6481 tcctattcta tttgaagtat aggaacttca aatacaaggg cttactgtgc ggactgggac
     6541 gcccccccct ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     6601 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     6661 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     6721 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     6781 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     6841 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     6901 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     6961 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     7021 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa acgtctaggc cccccgaacc
     7081 acggggacgt ggttttcctt tgaaaaacac gatgataata tggccaccac catggtggtc
     7141 aaggtgctgt tcgccctgat ctgtattgcc gtggccgagg ccaagcctac cgtgttcacc
     7201 ctggaagatt tcgtcggcga ttggagacag accgccggat acaacctgga ccaggtgttg
     7261 gagcaaggcg gcgtgtccag cctgtttcag aacctgggag tgtccgtgac acccatccag
     7321 agaatcgtgc tgagcggcga gaacggcctg aagatcgaca tccatgtgat catcccttac
     7381 gagggcctgt ccggcgatca gatgggccag atcgagaaga ttttcaaggt ggtgtacccc
     7441 gtggacgacc accacttcaa agtgatcctg cactacggca ccctggtcat cgatggcgtg
     7501 acccctaaca tgatcgacta cttcggcaga ccctacgagg gaatcgccgt gttcgacggc
     7561 aagaaaatca ccgtgaccgg cacactgtgg aacggcaaca agatcatcga cgagcggctg
     7621 atcaaccccg atggcagcct gctgttcaga gtgaccatca acggcgtgac aggatggcgg
     7681 ctgtgcgaaa gaatcctggc ctaatgtaca gatatcacta tcgataccgc ggggatcctc
     7741 tagagtagac cacctcccct gcgagctaag ctggacagcc aatgacgact aagagagtga
     7801 catttttcac taagctaaga caggagggcc gtcagagcta ctgcctaatc caaagacggc
     7861 taaaagtgat aaaaatgtat cactccaagt taagacaggc gcagcttccg agggatttgc
     7921 ctggccggat tgactgacgt cttaaccact gcatatcgat accgcgggga tcctctagag
     7981 tagaccacct cccctgcgag ctaagctgga cagccaatga cgactaagag agtgacattt
     8041 ttcactaagc taagacagga gggccgtcag agctactgcc taatccaaag acggctaaaa
     8101 gtgataaaaa tgtatcactc caagttaaga caggcgcagc ttccgaggga tttgcgttta
     8161 aacgcggccg ctctatactc gacgattgac tatctgacaa atcatagaga acatagagaa
     8221 cctcctcgag tcgcctgctt ttctgcctgg tactaacttc tctcccctct cctcttttct
     8281 ttttctgcag gaagtgctaa atttagctca aagcaaaaac tttcacttaa gacccaggga
     8341 cttaatcagc aatatcaacg taatagttct ggaactaaag gtaaggcatt actttatttg
     8401 ctctcctgga aataaaaaaa aaaaagtagg gggaaaagta ccacatttta aagtgacata
     8461 acatttttgg tatttgtaaa gtacccatgc atgtaattag cctacatttt aagtacactg
     8521 tgaacatgaa tcatttctaa tgttaaatga ttaactgggg agtataagct actgagtttg
     8581 cacctaccat ctactaatgg acaagcctca tcccaaactc catcaccttt catattaaca
     8641 caaaactggg agtgagagaa ggtactgagt tgagtttcac agaaagcagg cagattttat
     8701 tatatatttt tcaattcctt cagatcattt actggaatag ccaatactga ttacctgaaa
     8761 ggcttttcaa atggtgtttc cttatcattt gatggaagga ctacccataa gagatttgtc
     8821 ttaaaaaaaa aaactggagc cattaaaatg gccagtggac taaacaaaca acaatctttt
     8881 tagaggcaat ccccactttc agaatcttaa gtatttttaa atgcacagga agcataaaat
     8941 atgcaaggga ctcaggtgat gtaaaagaga ttcacttttg tctttttata tcccgtctcc
     9001 taaggtataa aattcatgag ttaataggta tcctaaataa gcagcataag tatagtagta
     9061 aaagacattc ctaaaagtaa ctccagttgt gtccaaatga atcacttatt agtggactgt
     9121 ttcagttgaa ttaaaaaaat acattgagat caatgtcatc tagacattga cagattcagt
     9181 tccttatcta tggcaagagt tttactctaa aataattaac atcagaaaac tcattcttaa
     9241 ctcttgatac aaatttaaga caaaaccatg caaaaatctg aaaactgtgt ttcaaaagcc
     9301 aaacactttt taaaataaaa aaatcccaag atatgacaat atttaaacaa ttatgcttaa
     9361 gaggatacag aacactgcaa cagtttttta aaagagaata cttatttaaa gggaacactc
     9421 tatctcacct gcttttgttc ccagggtagg aatcacttca aatttgaaaa gctctctttt
     9481 aaatctcact atatatcaaa atatttcctc cttagcttat caactagagg aagcgtttaa
     9541 atagctcctt tcagcagaga agcctaattt ctaaaaagcc agtccacaga acaaaatttc
     9601 taatgtttaa acttttaaaa gttggcaaat tcacctgcat tgatactatg atggggtagg
     9661 gataggtgta agtatttatg aagatgttct tcacacaaat ttatcccaaa cagaagcatg
     9721 tcctagctta ctctagtgta gttctgttct gctttgggga aaatataagg agattcactt
     9781 aagtagaaaa ataggagact ctaatcaaga tttagaaaag aagaaagtat aatgtgcata
     9841 tcaattcata catttaactt acacaaatat aggtgtacat tcagaggaaa agcgatcaag
     9901 tttatttcac atccagcatt taatatttgt ctagatctat ttttatttaa atctttattt
     9961 gcacccaatt tagggaaaaa atttttgtgt tcattgactg aattaacaaa tgaggaaaat
    10021 ctcagcttct gtgttactat catttggtat cataacaaaa tatgtaattt tggcattcat
    10081 tttgatcatt tcaagaaaat gtgaataatt aatatgtttg gtaagcttga aaataaaggc
    10141 aacaggccta taagacttca attgggaata actgtatata aggtaaacta ctctgtactt
    10201 taaaaaatta acatttttct tttataggga tctgaaacaa cattcatgtg tgaatatgct
    10261 gatgagacag caaccattgt agaatttctg aacagatgga ttaccttttg tcaaagcatc
    10321 atctcaacac tgacttgata attaagtgct tcccacttaa aacatatcag gccttctatt
    10381 tatttaaata tttaaatttt atatttattg ttgaatgtat ggtttgctac ctattgtaac
    10441 tattattctt aatcttaaaa ctataaatat ggatctttta tgattctttt tgtaagccct
    10501 aggggctcta aaatggtttc acttatttat cccaaaatat ttattattat gttgaatgtt
    10561 aaatatagta tctatgtaga ttggttagta aaactattta ataaatttga taaatataaa
    10621 caagcctgga tatttgttat tttggaaaca gcacagagta agcatttaaa tatttcttag
    10681 ttacttgtgt gaactgtagg atggttaaaa tgcttacaaa agtcactctt tctctgaaga
    10741 aatatgtaga acagagatgt agacttctca aaagcccttg ctttgtcctt tcaagggctg
    10801 atcagaccct tagttctggc atctcttagc agattatatt ttccttcttc ttaaaatgcc
    10861 aaacacaaac actcttgaaa ctcttcatag atttggtgtg gctatgaatt ctccaatatc
    10921 ttacaccctg cccagtgctg tgaggaggct cacctgtatg gcctatatca aaggtcttcc
    10981 ctgccctttg gctttccatt gggtcctgcc actggggagt gctggtagga actatgagga
    11041 acataagaga ttcccttgac tccctccttg tggagtagac ccaggatggc tgtgtctctc
    11101 aagcaaggaa cccagattac ctcaaggtgg cactctgggt actttttcct tctgagtgat
    11161 tctggtaatc ttcggagacg gtacccagct tttgttccct ttagtgaggg ttaattgcgc
    11221 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
    11281 cacacaacat acgagccggg agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
    11341 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
    11401 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
    11461 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
    11521 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
    11581 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
    11641 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
    11701 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
    11761 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
    11821 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
    11881 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
    11941 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
    12001 acaggattag cagagcgagg tatgtaggag gtgctacaga gttcttgaag tggtggccta
    12061 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
    12121 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
    12181 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
    12241 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
    12301 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
    12361 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
    12421 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
    12481 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
    12541 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
    12601 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
    12661 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
    12721 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
    12781 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
    12841 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
    12901 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
    12961 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
    13021 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
    13081 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
    13141 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
    13201 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
    13261 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
    13321 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
    13381 tgccac
//
