LOCUS       sequence-423952-        8761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luminescent reporter system to measure EJC-enhanced NMD activity. No
            intron in 5'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8761)
  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 8761)
  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. 2024 Feb 20:gkae121. doi: 10.1093/nar/gkae121."
COMMENT     Sequence Label: LRTi
FEATURES             Location/Qualifiers
     source          1..8761
                     /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"
     regulatory      3316..3325
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3325..3354
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3355..5001
                     /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(3418..3436)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             5056..5121
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5122..5175
                     /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             5176..5217
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6019..6045
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    6186..6307
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    6403..6514
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6503..6520
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(6656..7255)
                     /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(6739..6759)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7236..7255
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     protein_bind    complement(7260..7293)
                     /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    7505..7553
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        7626..7730
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7731..7799,7800..8591)
                     /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(7949..7968)
                     /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 ataccggtcg ccaccgccac catggaacaa aagctgattt ctgaagaaga cttggaagac
     3361 gccaaaaaca taaagaaagg cccggcgcca ttctatccgc tggaagatgg aaccgctgga
     3421 gagcaactgc ataaggctat gaagagatac gccctggttc ctggaataat tgcttttaca
     3481 gatgcacata tcgaggtgga catcacttac gctgagtact tcgaaatgtc cgttcggttg
     3541 gcagaagcta tgaaacgata tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa
     3601 aactctcttc aattctttat gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg
     3661 cccgcgaacg acatttataa tgaacgtgaa ttgctcaaca gtatgggcat ttcgcagcct
     3721 accgtggtgt tcgtttccaa aaaggggttg caaaaaattt tgaacgtgca aaaaaagctc
     3781 ccaatcatcc aaaaaattat tatcatggat tctaaaacgg attaccaggg atttcagtcg
     3841 atgtacacgt tcgtcacatc tcatctacct cccggtttta atgaatacga ttttgtgcca
     3901 gagtccttcg atagggacaa gataattgca ctgatcatga actcctctgg atctactggt
     3961 ctgcctaaag gtgtcgctct gcctcataga actgcctgcg tgagattctc gcatgccaga
     4021 gatcctattt ttggcaatca aatcattccg gatactgcga ttttaagtgt tgttccattc
     4081 catcacggtt ttggaatgtt tactacactc ggatatttga tatgtggatt tcgagtcgtc
     4141 ttaatgtata gatttgaaga agagctgttt ctgaggagcc ttcaggatta caagattcaa
     4201 agtgcgctgc tggtgccaac cctattctcc ttcttcgcca aaagcactct gattgacaaa
     4261 tacgatttat ctaatttaca cgaaattgct tctggtggcg ctcccctctc taaggaagtc
     4321 ggggaagcgg ttgccaagag gttccatctg ccaggtatca ggcaaggata tgggctcact
     4381 gagactacat cagctattct gattacaccc gagggggatg ataaaccggg cgcggtcggt
     4441 aaagttgttc cattttttga agcgaaggtt gtggatctgg ataccgggaa aacgctgggc
     4501 gttaatcaaa gaggcgaact gtgtgtgaga ggtcctatga ttatgtccgg ttatgtaaac
     4561 aatccggaag cgaccaacgc cttgattgac aaggatggat ggctacattc tggagacata
     4621 gcttactggg acgaagacga acacttcttc atcgttgacc gcctgaagtc tctgattaag
     4681 tacaaaggct atcaggtggc tcccgctgaa ttggaatcca tcttgctcca acaccccaac
     4741 atcttcgacg caggtgtcgc aggtcttccc gacgatgacg ccggtgaact tcccgccgcc
     4801 gttgttgttt tggagcacgg aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc
     4861 agtcaagtaa caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg
     4921 aaaggtctta ccggaaaact cgacgcaaga aaaatcagag agatcctcat aaaggccaag
     4981 aagggcggaa agatcgccgt gctcgagggt ggttctggtg gttctgccta aggtggttct
     5041 ggtggttctg aattcgacta caaggaccac gacggtgact acaaggacca cgacatcgac
     5101 tacaaggacg acgacgacaa ggagggcaga ggaagtctgc taacatgcgg tgacgtcgag
     5161 gagaatcctg gcccaggtaa gcctatccct aaccctctcc tcggtctcga ttctacgagc
     5221 gggggcgagg agctgttcgc cggcatcgtg cccgtgctga tcgagctgga cggcgacgtg
     5281 cacggccaca agttcagcgt gcgcggcgag ggcgagggcg acgccgacta cggcaagctg
     5341 gagatcaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctggtgacc
     5401 accctctgct acggcatcca gtgcttcgcc cgctaccccg agcacatgaa gatgaacgac
     5461 ttcttcaaga gcgccatgcc cgagggctac atccaggagc gcaccatcca gttccaggac
     5521 gacggcaagt acaagacccg cggcgaggtg agtgctacat ggtgagcccc aaagctggtg
     5581 tgggaggagc cacctggctg atgggcagcc ccttcatacc ctcacgtatt cccccaggtg
     5641 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcaagga cttcaaggag
     5701 gacggcaaca tcctgggcca caagctggag tacagcttca acagccacaa cgtgtacatc
     5761 cgccccgaca aggccaacaa cggcctggag gctaacttca agacccgcca caacatcgag
     5821 ggcggcggcg tgcagctggc cgaccactac cagaccaacg tgcccctggg cgacggcccc
     5881 gtgctgatcc ccatcaacca ctacctgagc actcagacca agatcagcaa ggaccgcaac
     5941 gaggcccgcg accacatggt gctcctggag tccttcagcg cctgctgcca cacccacggc
     6001 atggacgagc tgtacaggta cccatacgat gttccagatt acgctatgca ttaaatagat
     6061 ccaccggatc tagataactg atcataatca gccataccac atttgtagag gttttacttg
     6121 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     6181 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6241 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     6301 tatcttaacg cgaaactagt gctattgtct tcccaatcct cccccttgct gtcctgcccc
     6361 accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt
     6421 tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg
     6481 caaacaacag atggctggca actagaaggc acagtcgagg ctgatcagcg agctctagag
     6541 tcggtgggcc tcgggggcgg gtgcggggtc ggcggggccg ccccgggtgg cttcggtcgg
     6601 agccatgggg tcgtgcgctc ctttcggtcg ggcgctgcgg gtcgtggggc gggcgtcagg
     6661 caccgggctt gcgggtcatg caccaggtgc gcggtccttc gggcacctcg acgtcggcgg
     6721 tgacggtgaa gccgagccgc tcgtagaagg ggaggttgcg gggcgcggag gtctccagga
     6781 aggcgggcac cccggcgcgc tcggccgcct ccactccggg gagcacgacg gcgctgccca
     6841 gacccttgcc ctggtggtcg ggcgacacgc cgacggtggc caggaaccac gcgggctcct
     6901 tgggccggtg cggcgccagg aggccttcca tctgttgctg cgcggccagc cgggaaccgc
     6961 tcaactcggc catgcgcggg ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg
     7021 gcgtggtcca gaccgccacc gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga
     7081 gcccgacgcg cgtgaggaag agttcttgca gctcggtgac ccgctcgatg tggcggtccg
     7141 ggtcgacggt gtggcgcgtg gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg
     7201 cccgggggac gtcgtcgcgg gtggcgaggc gcaccgtggg cttgtactcg gtcatggtga
     7261 taacttcgta taggatactt tatacgaagt tatttattct cgaatggggg accccgtccc
     7321 taacccacgg ggccagtggc tatggcaggg cctgccgccc cgacgttggc tgcgagccct
     7381 gggccttcac ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg
     7441 ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt
     7501 ttatgaacaa acgacccaac acccgtgcgt tttattctgt ctttttattg ccgtcatagc
     7561 gcgggttcct tccggtattg tctccttccg tgtttattaa gcttggcact tttcggggaa
     7621 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     7681 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     7741 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     7801 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     7861 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     7921 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     7981 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     8041 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     8101 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     8161 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     8221 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     8281 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     8341 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     8401 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     8461 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     8521 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     8581 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     8641 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     8701 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     8761 c
//
