LOCUS       sequence-489485-       17223 bp    DNA     circular SYN 16-OCT-2025
DEFINITION  PiggyBac transposon vector for expression of dsRedExpress2 under 
            synTF promoter; constitutive expression of IL-10Rb NTEVp chain, 
            IL-10Ra CTEVp chain, and mNeonGreen-P2A-PuroR selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17223)
  AUTHORS   Edelstein HI, Cosio A, Ezekiel ML, Corcoran WK, Morris AH, Leonard 
            JN
  TITLE     Conversion of natural cytokine receptors into orthogonal synthetic 
            biosensors.
  JOURNAL   Nat Chem Biol. 2025 Aug 22:10.1038/s41589-025-01986-1. doi: 
            10.1038/s41589-025-01986-1.
  PUBMED    40846994
REFERENCE   2  (bases 1 to 17223)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol. 2025 Aug 22:10.1038/s41589-025-01986-1. doi: 
            10.1038/s41589-025-01986-1."
COMMENT     Sequence Label: L4385 dsRedExpress2 reporter, IL-10Rb NTEVp, I...
FEATURES             Location/Qualifiers
     source          1..17223
                     /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     complement(737..756)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             748..1557
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(815..834)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     repeat_region   1651..1685
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     regulatory      2937..2946
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2943..3620
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(3123..3143)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     3535..3558
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    3671..3895
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        4944..6126
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5175..6117
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     regulatory      6233..6242
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6302..6367
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    7474..7698
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        8747..9929
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          8978..9920
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     CDS             10105..10170
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             11275..11295
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     CDS             11314..11334
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11341..11490
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     polyA_signal    11871..12095
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        13144..14326
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          13375..14317
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     regulatory      14427..14436
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      14688..14697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             15141..15197
                     /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             15198..15800
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     15697..15717
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    15851..16075
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   17015..17077
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
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 ctggagctag catggcccgt gtctcaaaat
      661 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
      721 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg
      781 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
      841 gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt
      901 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
      961 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     1021 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
     1081 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     1141 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     1201 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     1261 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     1321 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     1381 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     1441 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     1501 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     1561 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca
     1621 agctcatgac ggtcgccacc gctagattaa ccctagaaag atagtctgcg taaaattgac
     1681 gcatgcattc ttgaaatatt gctctctctt tctaaatagc gcgaatccgt cgctgtgcat
     1741 ttaggacatc tcagtcgccg cttggagctc ccgtgaggcg tgcttgtcaa tgcggtaagt
     1801 gtcactgatt ttgaactata acgaccgcgt gagtcaaaat gacgcatgat tatcttttac
     1861 gtgactttta agatttaact catacgataa ttatattgtt atttcatgtt ctacttacgt
     1921 gataacttat tatatatata ttttcttgtt atagatatca ttgatgagtt tggacaaacc
     1981 gcatcgcgat aagctctaga gggacagccc ccccccaaag cccccaggga tgtaattacg
     2041 tccctccccc gctagggggc agcagcgagc cgcccggggc tccgctccgg tccggcgctc
     2101 cccccgcatc cccgagccgg cagcgtgcgg ggacagcccg ggcacgggga aggtggcacg
     2161 ggatcgcttt cctctgaacg cttctcgctg ctctttgagc ctgcagacac ctggggggat
     2221 acggggaaaa agctttaggc tgaaagagag atttagaatg acagaatcat agaacggcct
     2281 gcgtacggat ggggctctag ctctagagct tgatatcgaa ttcctgcagc cccgctagag
     2341 ggacagcccc cccccaaagc ccccagggat gtaattacgt ccctcccccg ctagggggca
     2401 gcagcgagcc gcccggggct ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc
     2461 agcgtgcggg gacagcccgg gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc
     2521 ttctcgctgc tctttgagcc tgcagacacc tggggggata cggggaaaaa gctttaggct
     2581 gaaagagaga tttagaatga cagaatcata gaacggcctg ggcgcgcctg gtggcgaacg
     2641 atcaacagac gggagattgc caaggtcttg acaaacgagt gctagtacaa gatctgcgag
     2701 ggtgtagggg ttaatggtgt aggggtaatt ggtgtagggg tctagggtgt aggggtaatg
     2761 ggtgtagggg tttaaggtgt aggggtagca ggtgtagggg ttcgtggtgt aggggttata
     2821 ggtgtagggg taattggtgt aggggtatgt ggtgtagggg taacgggtgt aggggtacgc
     2881 tctctagagg gtatataatg ggggccacta gtctactacc agagctcatc gctagcgcca
     2941 ccatggatag cactgagaac gtcatcaagc ccttcatgcg cttcaaggtg cacatggagg
     3001 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggcaag ccctacgagg
     3061 gcacccagac cgccaagctc caggtgacca agggcggccc cctgcccttc gcctgggaca
     3121 tcctgtcccc ccagttccag tacggctcca aggtgtacgt gaagcacccc gccgacatcc
     3181 ccgactacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     3241 aggacggcgg cgtggtgacc gtgacccagg actcctccct ccaggacggc accttcatct
     3301 accacgtgaa gttcatcggc gtgaacttcc cctccgacgg ccccgtaatg cagaagaaaa
     3361 ctctgggctg ggagccctcc accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg
     3421 agatccacaa ggcgctgaag ctgaagggcg gcggccacta cctggtggag ttcaagtcaa
     3481 tctacatggc caagaagccc gtgaagctgc ccggctacta ctacgtggac tccaagctgg
     3541 acatcacctc ccacaacgag gactacaccg tggtggagca gtacgagcgc gccgaggccc
     3601 gccaccacct gttccagtag gcggccgctc gagtctagag ggcccgttta aacccgctga
     3661 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3721 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3781 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3841 ggggaggatt gggaagtcaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3901 gaggcaggtc gagggtacat gcgccttact ccttgtgagc gcatcgcgat aagctctaga
     3961 gggacagccc ccccccaaag cccccaggga tgtaattacg tccctccccc gctagggggc
     4021 agcagcgagc cgcccggggc tccgctccgg tccggcgctc cccccgcatc cccgagccgg
     4081 cagcgtgcgg ggacagcccg ggcacgggga aggtggcacg ggatcgcttt cctctgaacg
     4141 cttctcgctg ctctttgagc ctgcagacac ctggggggat acggggaaaa agctttaggc
     4201 tgaaagagag atttagaatg acagaatcat agaacggcct gcgtacggat ggggctctag
     4261 ctctagagct tgatatcgaa ttcctgcagc cccgctagag ggacagcccc cccccaaagc
     4321 ccccagggat gtaattacgt ccctcccccg ctagggggca gcagcgagcc gcccggggct
     4381 ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc agcgtgcggg gacagcccgg
     4441 gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc ttctcgctgc tctttgagcc
     4501 tgcagacacc tggggggata cggggaaaaa gctttaggct gaaagagaga tttagaatga
     4561 cagaatcata gaacggcctg ggcgcgcctg gtggcgaacg atcaacagac gggagagggc
     4621 aaaaccgatg ttgacggact aatcctgaca agatctgggg aggtcgctga gtagtgcgcg
     4681 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     4741 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg tgagtaattc
     4801 atacaaaagg actcgcccct gccttgggga atcccaggga ccgtcgttaa actcccacta
     4861 acgtagaacc cagagatcgc tgcgttcccg ccccctcacc cgcccgctct cgtcatcact
     4921 gaggtggaga agagcatgcg tgaggctccg gtgcccgtca gtgggcagag cgcacatcgc
     4981 ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt
     5041 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
     5101 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
     5161 ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt
     5221 atggcccttg cgtgccttga attacttcca cgcccctggc tgcagtacgt gattcttgat
     5281 cccgagcttc gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct
     5341 tcgcctcgtg cttgagttga ggcctggctt gggcgctggg gccgccgcgt gcgaatctgg
     5401 tggcaccttc gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga
     5461 tgacctgctg cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg
     5521 cacactggta tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc
     5581 acatgttcgg cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct
     5641 caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg
     5701 gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc
     5761 cctgctgcag ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca
     5821 cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag
     5881 taccgggcgc cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta
     5941 ggttgggggg aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa
     6001 gttaggccag cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga
     6061 tcttggttca ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg
     6121 tcgtgagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
     6181 acctccatag aagagaacgg gaccgatcca aattctcgag gctagctgta cagccaccat
     6241 ggccctgcct gtgacagccc tgctgctgcc tctggctctg ctgctgcacg ccgctagacc
     6301 cgactacaag gaccacgacg gcgattataa ggatcatgac atcgattaca aggacgatga
     6361 cgacaagatg gtgcctccac ctgagaacgt gcggatgaac agcgtgaact tcaagaacat
     6421 cctccagtgg gagagccccg ccttcgccaa gggaaatctg accttcacag cccagtacct
     6481 gagctaccgg atcttccagg acaagtgcat gaacaccaca ctgaccgagt gcgacttcag
     6541 cagcctgtct aagtacggcg atcacaccct gagagtgcgg gccgagtttg ccgatgagca
     6601 cagcgattgg gtcaacatca cattctgccc cgtggacgac accatcatcg gcccacctgg
     6661 aatgcaggtt gaggtgctgg ccgatagcct gcacatgaga tttctggccc ctaagatcga
     6721 gaacgagtac gagacatgga ccatgaagaa cgtctacaac agctggacct acaacgtgca
     6781 gtactggaag aacggcaccg acgagaagtt ccagatcacc cctcagtacg acttcgaggt
     6841 gctgcggaac ctggaacctt ggaccaccta ctgtgtgcaa gtgcggggct tcctgcctga
     6901 cagaaacaaa gctggcgagt ggagcgagcc tgtgtgcgag cagacaaccc acgatgagac
     6961 agtgccctct tggatggtgg ccgtgatcct gatggccagc gtgttcatgg tttgcctggc
     7021 tctgctgggc tgcttcgctc tgctttggtg cgtctacaag aagggcgccg agagcttgtt
     7081 taaggggccg cgtgattaca acccgatatc gagcaccatt tgtcatttga cgaatgaatc
     7141 tgatgggcac acaacatcgt tgtatggtat tggatttggt cccttcatca ttacaaacaa
     7201 gcacttgttt agaagaaata atggaacact gttggtccaa tcactacatg gtgtattcaa
     7261 ggtcaagaac accacgactt tgcaacaaag cctcattgat gggagggaca tgataattat
     7321 tcgcatgcct aaggatttcc caccatttcc tcaaaagctg aaatttagag agccacaaag
     7381 ggaagagcgc atatgtcttg tgacaaccaa cttccaaact taggcggccg ctcgagtcta
     7441 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     7501 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     7561 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     7621 gtggggtggg gcaggacagc aagggggagg attgggaagt caatagcagg catgctgggg
     7681 atgcggtggg ctctatggct tctgaggcag gtcgaagaac tatttcctgg ctgttacgcg
     7741 agcgcatcgc gataagctct agagggacag ccccccccca aagcccccag ggatgtaatt
     7801 acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc cggtccggcg
     7861 ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg ggaaggtggc
     7921 acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga cacctggggg
     7981 gatacgggga aaaagcttta ggctgaaaga gagatttaga atgacagaat catagaacgg
     8041 cctgcgtacg gatggggctc tagctctaga gcttgatatc gaattcctgc agccccgcta
     8101 gagggacagc ccccccccaa agcccccagg gatgtaatta cgtccctccc ccgctagggg
     8161 gcagcagcga gccgcccggg gctccgctcc ggtccggcgc tccccccgca tccccgagcc
     8221 ggcagcgtgc ggggacagcc cgggcacggg gaaggtggca cgggatcgct ttcctctgaa
     8281 cgcttctcgc tgctctttga gcctgcagac acctgggggg atacggggaa aaagctttag
     8341 gctgaaagag agatttagaa tgacagaatc atagaacggc ctgggcgcgc ctggtggcga
     8401 acgatcaaca gacgggagag gactaaatag tagttcagac gccgttaagc gcaagatctg
     8461 gggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     8521 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
     8581 ccagatatac gcgtgagtaa ttcatacaaa aggactcgcc cctgccttgg ggaatcccag
     8641 ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc ccgccccctc
     8701 acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct ccggtgcccg
     8761 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     8821 ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     8881 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     8941 cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg
     9001 cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacgcccct
     9061 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
     9121 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cttgggcgct
     9181 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
     9241 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
     9301 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
     9361 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
     9421 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
     9481 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     9541 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     9601 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     9661 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     9721 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     9781 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     9841 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     9901 gtttttttct tccatttcag gtgtcgtgag agctcgttta gtgaaccgtc agatcgcctg
     9961 gagacgccat ccacgctgtt ttgacctcca tagaagagaa cgggaccgat ccaaattctc
    10021 gaggctagct gtacagccac catgctgccc tgcctggttg ttctgcttgc cgctctgctg
    10081 tctctgagac tgggatctga cgccgactac aaggaccacg acggcgatta taaggatcat
    10141 gacatcgatt acaaggacga tgacgacaag cacggcaccg agctgccttc tccacctagc
    10201 gtttggttcg aggccgagtt cttccaccac atcctgcact ggacccctat tcctaaccag
    10261 agcgagagca cctgttacga ggtggccctg ctgagatacg gcatcgagag ctggaacagc
    10321 atcagcaact gcagccagac actgagctac gacctgaccg ccgtgacact ggatctgtac
    10381 cacagcaacg gctaccgggc cagagttaga gccgtggatg gaagcagaca ctccaactgg
    10441 accgtgacca acaccagatt cagcgtggac gaagtgaccc tgacagtggg cagcgtgaac
    10501 ctggaaatcc acaacggctt catcctgggc aagatccagc tgcctcggcc taagatggcc
    10561 cctgccaatg atacctacga gagcatcttc agccacttcc gcgagtacga gatcgccatc
    10621 agaaaggtgc ccggcaactt caccttcaca cacaagaaag tgaagcacga gaacttcagc
    10681 ctgctgacct ctggcgaagt gggcgagttc tgcgtgcaag tgaaaccctc tgtggccagc
    10741 cggtccaaca aaggcatgtg gtccaaagag gaatgcatca gcctgaccag acagtacttc
    10801 accgtgacaa acgtgatcat cttcttcgcc ttcgtgctgc tgctgtctgg cgctctggct
    10861 tattgtctgg ccctgcagct gtacgtgcgg cggagaaaga agggcgccaa gagcatgtct
    10921 agcatggtgt cagacactag ttgcacattc ccttcatctg atggcatatt ctggaagcat
    10981 tggattcaaa ccaaggatgg gcagtgtggc agtccattag tatcaactag agatgggttc
    11041 attgttggta tacactcagc atcgaatttc accaacacaa acaattattt cacaagcgtg
    11101 ccgaaaaact tcatggaatt gaaaacaaat caggaggcgc agcagtgggt tagtggttgg
    11161 cgattaaatg ctgactcagt attgtggggg ggccataaag ttttcatggt gaaacctgaa
    11221 gagccttttc agccagttaa ggaagcgact caactcatga atcttaaggg ctcggagaac
    11281 ctgtattttc agatgggggg aggcagcggg ggccctaaga aaaagcgcaa agtctccgga
    11341 gacgctctgg atgactttga tctggacatg cttggttccg atgccttgga cgattttgac
    11401 ctcgatatgc tgggctccga cgctctcgac gatttcgacc tcgacatgct gggtagcgat
    11461 gcactggatg actttgatct ggatatgctc ggatccggtg gtggcgggtc tggaggagga
    11521 ggtagtggtg gaggtggctc tggtaccgct agacccggcg aaagaccttt ccagtgccgg
    11581 atctgcatga ggaacttcag caagggcgag agactcgtgc ggcacaccag aacacacaca
    11641 ggcgagaagc ccttccagtg tagaatctgt atgcgcaact tcagccggat ggacaacctg
    11701 agcacccacc tgagaaccca taccggggag aagccatttc aatgccgcat ctgtatgaga
    11761 aatttttccc ggaaggacgc cctgaaccgg cacctgaaaa cacacctgag aggcagctga
    11821 gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
    11881 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
    11941 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
    12001 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagtcaa
    12061 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcaggtc gaagagactg
    12121 cccgacacat cttagtgagc gcatcgcgat aagctctaga gggacagccc ccccccaaag
    12181 cccccaggga tgtaattacg tccctccccc gctagggggc agcagcgagc cgcccggggc
    12241 tccgctccgg tccggcgctc cccccgcatc cccgagccgg cagcgtgcgg ggacagcccg
    12301 ggcacgggga aggtggcacg ggatcgcttt cctctgaacg cttctcgctg ctctttgagc
    12361 ctgcagacac ctggggggat acggggaaaa agctttaggc tgaaagagag atttagaatg
    12421 acagaatcat agaacggcct gcgtacggat ggggctctag ctctagagct tgatatcgaa
    12481 ttcctgcagc cccgctagag ggacagcccc cccccaaagc ccccagggat gtaattacgt
    12541 ccctcccccg ctagggggca gcagcgagcc gcccggggct ccgctccggt ccggcgctcc
    12601 ccccgcatcc ccgagccggc agcgtgcggg gacagcccgg gcacggggaa ggtggcacgg
    12661 gatcgctttc ctctgaacgc ttctcgctgc tctttgagcc tgcagacacc tggggggata
    12721 cggggaaaaa gctttaggct gaaagagaga tttagaatga cagaatcata gaacggcctg
    12781 ggcgcgcctg gtggcgaacg atcaacagac gggagaggtt acaatatccc gtgaagcttg
    12841 agtggaatca agatctgggg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
    12901 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
    12961 gcttcgcgat gtacgggcca gatatacgcg tgagtaattc atacaaaagg actcgcccct
    13021 gccttgggga atcccaggga ccgtcgttaa actcccacta acgtagaacc cagagatcgc
    13081 tgcgttcccg ccccctcacc cgcccgctct cgtcatcact gaggtggaga agagcatgcg
    13141 tgaggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
    13201 gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa
    13261 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
    13321 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt
    13381 gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga
    13441 attacttcca cgcccctggc tgcagtacgt gattcttgat cccgagcttc gggttggaag
    13501 tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga
    13561 ggcctggctt gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct
    13621 cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt
    13681 tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt
    13741 tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg
    13801 cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct
    13861 ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc
    13921 ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa
    13981 atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc
    14041 ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca
    14101 cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta
    14161 tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt
    14221 gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc
    14281 tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgagagc tcgtttagtg
    14341 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagagaacgg
    14401 gaccgatcca aattctcgag gctagcgcca ccatggtttc caagggcgaa gaggacaata
    14461 tggccagcct gcctgccaca cacgagctgc acatctttgg cagcatcaac ggcgtggact
    14521 tcgacatggt cggacaaggc accggcaatc ccaacgacgg ctacgaggaa ctgaacctga
    14581 agtccacaaa gggcgacctc cagttcagcc cctggattct ggtgcctcac atcggctacg
    14641 gcttccacca gtacctgcct tatcctgacg gcatgagccc tttccaggcc gccatggttg
    14701 atggctctgg ctaccaggtc caccggacca tgcagtttga ggatggcgct tccctgaccg
    14761 tgaactacag atacacctac gagggcagcc acatcaaagg cgaggcccaa gtgaaaggca
    14821 caggcttccc tgctgatggc cccgtgatga ccaattctct gacagccgcc gactggtgca
    14881 gaagcaagaa aacatacccc aacgacaaga ccatcatcag caccttcaag tggtcctaca
    14941 ccaccggcaa cggcaagcgg tacagaagca ccgccagaac cacctacacc ttcgccaaac
    15001 ctatggccgc caactacctg aagaaccagc ctatgtacgt gttccgcaag accgagctga
    15061 agcactccaa gacagaactg aacttcaaag agtggcagaa agccttcacc gacgtgatgg
    15121 gcatggacga actctacaaa gccacgaact tcagcctgct gaagcaggct ggagacgtgg
    15181 aggagaaccc tggacccatg gccaccgagt acaagcccac ggtgcgcctc gccacccgcg
    15241 acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
    15301 gccacaccgt cgacccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
    15361 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
    15421 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggctcgc
    15481 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
    15541 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
    15601 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
    15661 ctggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
    15721 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
    15781 cccgcaagcc cggtgcctga gcggccgctc gagtctagag ggcccgttta aacccgctga
    15841 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
    15901 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
    15961 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
    16021 ggggaggatt gggaagtcaa tagcaggcat gctggggatg cggtgggctc tatggcttct
    16081 gaggcaggtc gacactgttt aacaagcccg tcagtagagc gcatcgcgat aagctctaga
    16141 gggacagccc ccccccaaag cccccaggga tgtaattacg tccctccccc gctagggggc
    16201 agcagcgagc cgcccggggc tccgctccgg tccggcgctc cccccgcatc cccgagccgg
    16261 cagcgtgcgg ggacagcccg ggcacgggga aggtggcacg ggatcgcttt cctctgaacg
    16321 cttctcgctg ctctttgagc ctgcagacac ctggggggat acggggaaaa agctttaggc
    16381 tgaaagagag atttagaatg acagaatcat agaacggcct gcgtacggat ggggctctag
    16441 ctctagagct tgatatcgaa ttcctgcagc cccgctagag ggacagcccc cccccaaagc
    16501 ccccagggat gtaattacgt ccctcccccg ctagggggca gcagcgagcc gcccggggct
    16561 ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc agcgtgcggg gacagcccgg
    16621 gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc ttctcgctgc tctttgagcc
    16681 tgcagacacc tggggggata cggggaaaaa gctttaggct gaaagagaga tttagaatga
    16741 cagaatcata gaacggcctg ggcgcgcctg gtggcgaacg atcaacagac gggagaggca
    16801 gagaggatgc acatgtgacc gagggatcca gatgctagtt aaaagttttg ttactttata
    16861 gaagaaattt tgagtttttg ttttttttta ataaataaat aaacataaat aaattgtttg
    16921 ttgaatttat tattagtatg taagtgtaaa tataataaaa cttaatatct attcaaatta
    16981 ataaataaac ctcgatatac agaccgataa aacacatgcg tcaattttac gcatgattat
    17041 ctttaacgta cgtcacaata tgattatctt tctagggtta atctagtata cgcgtgagca
    17101 aaaggtctga cgagaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
    17161 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
    17221 ttc
//
