LOCUS       sequence-423949-        8959 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Luminescent reporter system, readthrough control, NMD negative 
            control. Intron present in 5'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8959)
  AUTHORS   Kolakada D, Campbell AE, Galvis LB, Li Z, Lore M, Jagannathan S
  TITLE     A system of reporters for comparative investigation of 
            EJC-independent and EJC-enhanced nonsense-mediated mRNA decay.
  JOURNAL   Nucleic Acids Res. 2024 Feb 20:gkae121. doi: 10.1093/nar/gkae121.
  PUBMED    38375914
REFERENCE   2  (bases 1 to 8959)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2024 Feb 20:gkae121. doi: 10.1093/nar/gkae121."
COMMENT     Sequence Label: LiRT_RTctrl
FEATURES             Location/Qualifiers
     source          1..8959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    1049..1170
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(1183..1216)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1430..1449
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1448..1503
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1549..1568
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(1645..2577)
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="TSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAENA
                     VIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAWF
                     ELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDIA
                     LIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPREI
                     PLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKVK
                     GLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             complement(2578..2595)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      2601..2610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2643..2667)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     enhancer        2775..3078
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3079..3282
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3233..3253
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     ncRNA           3409..3435
                     /label=5' miR-155
                     /note="sequence upstream of the precursor of mouse miR-155 
                     microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     ncRNA           3461..3502
                     /label=3' miR-155
                     /note="sequence downstream of the precursor of mouse 
                     miR-155 microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     primer_bind     3577..3596
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      3610..3619
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3619..3648
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3649..5295
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="EDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGIIAFTDAH
                     IEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPA
                     NDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSM
                     YTFVTSHLPPGFNEYDFVPESFDRDKIIALIMNSSGSTGLPKGVALPHRTACVRFSHAR
                     DPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKI
                     QSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYG
                     LTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG
                     YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILL
                     QHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVF
                     VDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(3712..3730)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             5350..5415
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5416..5469
                     /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             5470..5511
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5512..6222
                     /codon_start=1
                     /product="monomeric green fluorescent protein, also known 
                     as mTagGFP"
                     /label=TagGFP2
                     /note="mammalian codon-optimized"
                     /translation="SGGEELFAGIVPVLIELDGDVHGHKFSVRGEGEGDADYGKLEIKF
                     ICTTGKLPVPWPTLVTTLCYGIQCFARYPEHMKMNDFFKSAMPEGYIQERTIQFQDDGK
                     YKTRGEVKFEGDTLVNRIELKGKDFKEDGNILGHKLEYSFNSHNVYIRPDKANNGLEAN
                     FKTRHNIEGGGVQLADHYQTNVPLGDGPVLIPINHYLSTQTKISKDRNEARDHMVLLES
                     FSACCHTHGMDELYR"
     CDS             6223..6249
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    6384..6505
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    6601..6712
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6701..6718
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(6854..7453)
                     /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(6937..6957)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7434..7453
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     protein_bind    complement(7458..7491)
                     /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    7703..7751
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        7824..7928
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7929..7997,7998..8789)
                     /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     complement(8147..8166)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tgggtaccta agatacattg atgagtttgg acaaaccaca
     1081 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     1141 gtaaccatta taagctgcaa taaacaagtt gacggtatcg taataacttc gtatagcata
     1201 cattatacga agttatatgg atccatatgc tagctcgacg ctcaggtcga gggatcttca
     1261 taagagaaga gggacagcta tgactgggag taaggaatgg acagcagggg gctgtttcat
     1321 atactgatga cctctttata gccacctttg ttcatggcag ccagcatatg gcatatgttg
     1381 ccaaactcta aaccaaatac tcattctgat gttttaaatg atttgccctc ccatatgtcc
     1441 ttccgagtga gagacacaaa aaattccaac acactattgc aatgaaaata aatttccttt
     1501 attagccaga agtcagatgc tcaaggggct tcatgatgtc cccataattt ttggcagagg
     1561 gaaaaagatc tcagtggtat ttgtgagcca gggcattggc cacaccagcc accaccttct
     1621 gataggcagc ctgcacctga gtgtttattg ttcatttttg agaactcgct caacgaacga
     1681 tttgatatat tttcccattt catcaggtgc atcttcttgc gaaaaatgaa gaccttttac
     1741 tttgacaaat tcagtattag gaaacttctt ggcaccttca acaatagcat tggaaaagaa
     1801 tcctgggtcc gattcaataa acatttttgg taaatcatca cttgcacgta gataagcatt
     1861 ataattccta acaatttgta caacgtcagg tttaccacct tttactaacg ggatttcacg
     1921 aggccatgat aatgttggac gacgaacttc acctttctct ttgaatggtt caagatatgc
     1981 tgcaaattct tctggttcta actttctcat gatttttgat ggcaacatgg tttccacgaa
     2041 gaagttattc tccaaaacca ttttttctcc ttcttcagat ttgatcaacg caatatcttc
     2101 ttcaatatca ggccattcat cccatgattc aatcacatct actacacttt cagcgtgaac
     2161 tattgctttg atcttatctt gatgctcata gctataatga aatgccaaac aagcacccca
     2221 atcatggccg acaaaaatga tcttctttgg taaattaaga agttcaaacc atgcagtaag
     2281 atatttgtaa tgatcaagta acctataaga accattacca gatttgcctg atttgcccat
     2341 accaataagg tctggtataa tacaccgcgc tactggctca atatgtggca caacatgtcg
     2401 ccataaataa gaagaggccg cgttaccatg taaaaaaata acagcatttt ctgcatgttt
     2461 ttctgaatca taataattaa taaatgaatc aagaacattc atttgtttac atctggccca
     2521 ccactgcgga ccagttatca tccgtttcct ttgttctgga tcataaactt tcgaagtgtg
     2581 gtgatggtga tgatgatcca ccatggtggc ggtggcgacc acgatcctca tcctgtctct
     2641 gcgatctgac ggttcactaa acgagctctg cttatatagg cctcccaccg tacacgccac
     2701 ctcgacatac tcagtagtta ttaatagtaa tcaattacgg ggtcattagt tcatagcgag
     2761 gctgcagtct accgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     2821 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     2881 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     2941 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     3001 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     3061 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     3121 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     3181 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     3241 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
     3301 ataccggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg
     3361 cttgtcgaga cagagaagac tcttgcgttt cggtcgactt ctagagctct ggaggcttgc
     3421 tgaaggctgt atgctagaga cgtacagatg cgtctcacag gacacaaggc ctgttactag
     3481 cactcacatg gaacaaatgg ccgaattcgg cgctagctgc tgatatcgca tacgcgtgga
     3541 ccagataggc acctattggt cttactgaca tccactttgc ctttctctcc acaggtgccg
     3601 gtcgccaccg ccaccatgga acaaaagctg atttctgaag aagacttgga agacgccaaa
     3661 aacataaaga aaggcccggc gccattctat ccgctggaag atggaaccgc tggagagcaa
     3721 ctgcataagg ctatgaagag atacgccctg gttcctggaa taattgcttt tacagatgca
     3781 catatcgagg tggacatcac ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa
     3841 gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag tgaaaactct
     3901 cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg
     3961 aacgacattt ataatgaacg tgaattgctc aacagtatgg gcatttcgca gcctaccgtg
     4021 gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc
     4081 atccaaaaaa ttattatcat ggattctaaa acggattacc agggatttca gtcgatgtac
     4141 acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt gccagagtcc
     4201 ttcgataggg acaagataat tgcactgatc atgaactcct ctggatctac tggtctgcct
     4261 aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc cagagatcct
     4321 atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc attccatcac
     4381 ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt cgtcttaatg
     4441 tatagatttg aagaagagct gtttctgagg agccttcagg attacaagat tcaaagtgcg
     4501 ctgctggtgc caaccctatt ctccttcttc gccaaaagca ctctgattga caaatacgat
     4561 ttatctaatt tacacgaaat tgcttctggt ggcgctcccc tctctaagga agtcggggaa
     4621 gcggttgcca agaggttcca tctgccaggt atcaggcaag gatatgggct cactgagact
     4681 acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt cggtaaagtt
     4741 gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct gggcgttaat
     4801 caaagaggcg aactgtgtgt gagaggtcct atgattatgt ccggttatgt aaacaatccg
     4861 gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga catagcttac
     4921 tgggacgaag acgaacactt cttcatcgtt gaccgcctga agtctctgat taagtacaaa
     4981 ggctatcagg tggctcccgc tgaattggaa tccatcttgc tccaacaccc caacatcttc
     5041 gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg aacttcccgc cgccgttgtt
     5101 gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt cgccagtcaa
     5161 gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt accgaaaggt
     5221 cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc caagaagggc
     5281 ggaaagatcg ccgtgctcga gggtggttct ggtggttctg gctacggtgg ttctggtggt
     5341 tctgaattcg actacaagga ccacgacggt gactacaagg accacgacat cgactacaag
     5401 gacgacgacg acaaggaggg cagaggaagt ctgctaacat gcggtgacgt cgaggagaat
     5461 cctggcccag gtaagcctat ccctaaccct ctcctcggtc tcgattctac gagcgggggc
     5521 gaggagctgt tcgccggcat cgtgcccgtg ctgatcgagc tggacggcga cgtgcacggc
     5581 cacaagttca gcgtgcgcgg cgagggcgag ggcgacgccg actacggcaa gctggagatc
     5641 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctggt gaccaccctc
     5701 tgctacggca tccagtgctt cgcccgctac cccgagcaca tgaagatgaa cgacttcttc
     5761 aagagcgcca tgcccgaggg ctacatccag gagcgcacca tccagttcca ggacgacggc
     5821 aagtacaaga cccgcggcga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     5881 ctgaagggca aggacttcaa ggaggacggc aacatcctgg gccacaagct ggagtacagc
     5941 ttcaacagcc acaacgtgta catccgcccc gacaaggcca acaacggcct ggaggctaac
     6001 ttcaagaccc gccacaacat cgagggcggc ggcgtgcagc tggccgacca ctaccagacc
     6061 aacgtgcccc tgggcgacgg ccccgtgctg atccccatca accactacct gagcactcag
     6121 accaagatca gcaaggaccg caacgaggcc cgcgaccaca tggtgctcct ggagtccttc
     6181 agcgcctgct gccacaccca cggcatggac gagctgtaca ggtacccata cgatgttcca
     6241 gattacgctt aaatagatcc accggatcta gataactgat cataatcagc cataccacat
     6301 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     6361 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     6421 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6481 tgtccaaact catcaatgta tcttaacgcg aaactagtgc tattgtcttc ccaatcctcc
     6541 cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc
     6601 gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag
     6661 gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct
     6721 gatcagcgag ctctagagtc ggtgggcctc gggggcgggt gcggggtcgg cggggccgcc
     6781 ccgggtggct tcggtcggag ccatggggtc gtgcgctcct ttcggtcggg cgctgcgggt
     6841 cgtggggcgg gcgtcaggca ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg
     6901 gcacctcgac gtcggcggtg acggtgaagc cgagccgctc gtagaagggg aggttgcggg
     6961 gcgcggaggt ctccaggaag gcgggcaccc cggcgcgctc ggccgcctcc actccgggga
     7021 gcacgacggc gctgcccaga cccttgccct ggtggtcggg cgacacgccg acggtggcca
     7081 ggaaccacgc gggctccttg ggccggtgcg gcgccaggag gccttccatc tgttgctgcg
     7141 cggccagccg ggaaccgctc aactcggcca tgcgcgggcc gatctcggcg aacaccgccc
     7201 ccgcttcgac gctctccggc gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc
     7261 acaccttgcc gatgtcgagc ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc
     7321 gctcgatgtg gcggtccggg tcgacggtgt ggcgcgtggc ggggtagtcg gcgaacgcgg
     7381 cggcgagggt gcgtacggcc cgggggacgt cgtcgcgggt ggcgaggcgc accgtgggct
     7441 tgtactcggt catggtgata acttcgtata ggatacttta tacgaagtta tttattctcg
     7501 aatgggggac cccgtcccta acccacgggg ccagtggcta tggcagggcc tgccgccccg
     7561 acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa
     7621 gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga gtgccagccc
     7681 tgggaccgaa ccccgcgttt atgaacaaac gacccaacac ccgtgcgttt tattctgtct
     7741 ttttattgcc gtcatagcgc gggttccttc cggtattgtc tccttccgtg tttattaagc
     7801 ttggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     7861 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     7921 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     7981 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     8041 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     8101 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     8161 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     8221 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     8281 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     8341 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     8401 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     8461 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     8521 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     8581 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     8641 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     8701 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     8761 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     8821 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     8881 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     8941 aaaagatcaa aggatcttc
//
