LOCUS       sequence-489486-       20394 bp    DNA     circular SYN 16-OCT-2025
DEFINITION  PiggyBac transposon vector for expression of dsRed-Express2 synTF 
            promoter; constitutive expression of VEGFR2 NTEVp chain, VEGFR1 
            CTEVp chain, and mNeonGreen-P2A-PuroR selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 20394)
  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 20394)
  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: L4857 dsRedExpress2 reporter, VEGFR2 NTEVp, VE...
FEATURES             Location/Qualifiers
     source          1..20394
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      957..966
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             963..1640
                     /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(1143..1163)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1555..1578
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1691..1915
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2964..4146
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3195..4137
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     CDS             4316..4381
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    7105..7329
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        8378..9560
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          8609..9551
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     regulatory      9667..9676
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             9751..9816
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             12466..12486
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     CDS             12505..12525
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12532..12681
                     /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    13062..13286
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        14335..15517
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          14566..15508
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     regulatory      15618..15627
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      15879..15888
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             16332..16388
                     /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             16389..16991
                     /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     16888..16908
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    17042..17266
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   18206..18268
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     rep_origin      18415..19003
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     18904..18923
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(19151..19170)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             19162..19971
                     /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(19229..19248)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     repeat_region   20065..20099
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 gcatcgcgat aagctctaga gggacagccc ccccccaaag cccccaggga tgtaattacg
       61 tccctccccc gctagggggc agcagcgagc cgcccggggc tccgctccgg tccggcgctc
      121 cccccgcatc cccgagccgg cagcgtgcgg ggacagcccg ggcacgggga aggtggcacg
      181 ggatcgcttt cctctgaacg cttctcgctg ctctttgagc ctgcagacac ctggggggat
      241 acggggaaaa agctttaggc tgaaagagag atttagaatg acagaatcat agaacggcct
      301 gcgtacggat ggggctctag ctctagagct tgatatcgaa ttcctgcagc cccgctagag
      361 ggacagcccc cccccaaagc ccccagggat gtaattacgt ccctcccccg ctagggggca
      421 gcagcgagcc gcccggggct ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc
      481 agcgtgcggg gacagcccgg gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc
      541 ttctcgctgc tctttgagcc tgcagacacc tggggggata cggggaaaaa gctttaggct
      601 gaaagagaga tttagaatga cagaatcata gaacggcctg ggcgcgcctg gtggcgaacg
      661 atcaacagac gggagattgc caaggtcttg acaaacgagt gctagtacaa gatctgcgag
      721 ggtgtagggg ttaatggtgt aggggtaatt ggtgtagggg tctagggtgt aggggtaatg
      781 ggtgtagggg tttaaggtgt aggggtagca ggtgtagggg ttcgtggtgt aggggttata
      841 ggtgtagggg taattggtgt aggggtatgt ggtgtagggg taacgggtgt aggggtacgc
      901 tctctagagg gtatataatg ggggccacta gtctactacc agagctcatc gctagcgcca
      961 ccatggatag cactgagaac gtcatcaagc ccttcatgcg cttcaaggtg cacatggagg
     1021 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggcaag ccctacgagg
     1081 gcacccagac cgccaagctc caggtgacca agggcggccc cctgcccttc gcctgggaca
     1141 tcctgtcccc ccagttccag tacggctcca aggtgtacgt gaagcacccc gccgacatcc
     1201 ccgactacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     1261 aggacggcgg cgtggtgacc gtgacccagg actcctccct ccaggacggc accttcatct
     1321 accacgtgaa gttcatcggc gtgaacttcc cctccgacgg ccccgtaatg cagaagaaaa
     1381 ctctgggctg ggagccctcc accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg
     1441 agatccacaa ggcgctgaag ctgaagggcg gcggccacta cctggtggag ttcaagtcaa
     1501 tctacatggc caagaagccc gtgaagctgc ccggctacta ctacgtggac tccaagctgg
     1561 acatcacctc ccacaacgag gactacaccg tggtggagca gtacgagcgc gccgaggccc
     1621 gccaccacct gttccagtag gcggccgctc gagtctagag ggcccgttta aacccgctga
     1681 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     1741 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     1801 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     1861 ggggaggatt gggaagtcaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     1921 gaggcaggtc gagggtacat gcgccttact ccttgtgagc gcatcgcgat aagctctaga
     1981 gggacagccc ccccccaaag cccccaggga tgtaattacg tccctccccc gctagggggc
     2041 agcagcgagc cgcccggggc tccgctccgg tccggcgctc cccccgcatc cccgagccgg
     2101 cagcgtgcgg ggacagcccg ggcacgggga aggtggcacg ggatcgcttt cctctgaacg
     2161 cttctcgctg ctctttgagc ctgcagacac ctggggggat acggggaaaa agctttaggc
     2221 tgaaagagag atttagaatg acagaatcat agaacggcct gcgtacggat ggggctctag
     2281 ctctagagct tgatatcgaa ttcctgcagc cccgctagag ggacagcccc cccccaaagc
     2341 ccccagggat gtaattacgt ccctcccccg ctagggggca gcagcgagcc gcccggggct
     2401 ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc agcgtgcggg gacagcccgg
     2461 gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc ttctcgctgc tctttgagcc
     2521 tgcagacacc tggggggata cggggaaaaa gctttaggct gaaagagaga tttagaatga
     2581 cagaatcata gaacggcctg ggcgcgcctg gtggcgaacg atcaacagac gggagagggc
     2641 aaaaccgatg ttgacggact aatcctgaca agatctgggg aggtcgctga gtagtgcgcg
     2701 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     2761 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg tgagtaattc
     2821 atacaaaagg actcgcccct gccttgggga atcccaggga ccgtcgttaa actcccacta
     2881 acgtagaacc cagagatcgc tgcgttcccg ccccctcacc cgcccgctct cgtcatcact
     2941 gaggtggaga agagcatgcg tgaggctccg gtgcccgtca gtgggcagag cgcacatcgc
     3001 ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt
     3061 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
     3121 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
     3181 ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt
     3241 atggcccttg cgtgccttga attacttcca cgcccctggc tgcagtacgt gattcttgat
     3301 cccgagcttc gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct
     3361 tcgcctcgtg cttgagttga ggcctggctt gggcgctggg gccgccgcgt gcgaatctgg
     3421 tggcaccttc gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga
     3481 tgacctgctg cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg
     3541 cacactggta tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc
     3601 acatgttcgg cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct
     3661 caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg
     3721 gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc
     3781 cctgctgcag ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca
     3841 cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag
     3901 taccgggcgc cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta
     3961 ggttgggggg aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa
     4021 gttaggccag cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga
     4081 tcttggttca ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg
     4141 tcgtgagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
     4201 acctccatag aagagaacgg gaccgatcca aattctcgag gctagctgta cagccaccat
     4261 gcagagcaag gtgctgctgg ctgttgccct gtggctgtgc gtggaaacca gagccgacta
     4321 caaggaccac gacggcgatt ataaggatca cgacatcgat tacaaagacg acgacgacaa
     4381 ggccagcgtg ggcctgcctt ctgtttctct ggatctgccc agactgagca tccagaagga
     4441 catcctgacc atcaaggcca acaccacact ccagatcacc tgtcggggac agcgcgatct
     4501 ggattggctg tggcctaaca atcagagcgg cagcgagcag cgcgtggaag tgacagagtg
     4561 ttccgatggc ctgttctgca agaccctgac aatccccaaa gtgatcggca acgacaccgg
     4621 ggcctacaag tgcttctaca gagagacaga cctggcctcc gtgatctacg tgtacgtgca
     4681 ggactacaga agccccttta tcgccagcgt gtccgatcag cacggcgtgg tgtatatcac
     4741 cgagaacaag aacaagaccg tggtcatccc ctgcctgggc agcatcagca acctgaatgt
     4801 gtccctgtgc gcccggtatc ctgagaagag attcgtgccc gacggcaaca gaatcagctg
     4861 ggacagcaag aagggcttca ctatccccag ctacatgatc agctacgccg gcatggtgtt
     4921 ctgcgaggcc aagatcaacg acgagagcta ccagagcatc atgtatatcg tggtggtcgt
     4981 gggctaccgc atctacgatg tggtgctgag ccctagccac ggcatcgaac tgtctgtggg
     5041 agagaagctg gtgctgaact gcaccgccag aaccgagctg aatgtgggca tcgacttcaa
     5101 ctgggagtac cccagcagca agcaccagca caagaaactg gtcaaccggg acctgaaaac
     5161 ccagagcggc tccgagatga agaagttcct gagcacactg acaatcgacg gcgtgaccag
     5221 aagcgaccag ggcctgtata cttgtgccgc ctctagcggc ctgatgacca agaaaaacag
     5281 caccttcgtg cgggtccacg agaagccttt tgtggccttt ggcagcggca tggaaagcct
     5341 ggtggaagcc acagtgggcg agagagtcag aatccccgct aagtacctgg gctaccctcc
     5401 acctgagatc aagtggtaca agaacggcat ccctctggaa agcaaccaca caatcaaggc
     5461 cggccacgtg ctgacaatca tggaagtgtc cgagagagac accggcaatt acaccgtgat
     5521 cctgactaac cccatcagca aagaaaagca gagccacgtg gtgtccctgg tggtgtatgt
     5581 gcctcctcag atcggcgaga agtccctgat cagccccgtg gacagctacc agtacggcac
     5641 cacacagaca ctgacctgca ccgtgtacgc cattcctcca cctcaccaca tccactggta
     5701 ctggcagctg gaagaggaat gcgccaacga gccttctcag gccgtgtccg tgacaaaccc
     5761 ctatccttgt gaagagtggc ggagcgtgga agatttccaa ggcggcaaca agatcgaagt
     5821 caacaagaac cagttcgccc tgatcgaggg caagaacaaa acagtgtcta ccctcgtgat
     5881 ccaggccgcc aatgtgtctg ccctgtataa gtgcgaggcc gtgaacaaag tcggcagagg
     5941 cgaaagagtg atcagcttcc acgtgacaag aggccccgag atcaccctcc agccagatat
     6001 gcagcctacc gagcaagagt ccgtcagcct gtggtgtacc gccgacagat ccaccttcga
     6061 gaacctgacc tggtacaagc tgggacccca gcctctgcct attcacgtgg gagaactgcc
     6121 cacacctgtg tgcaagaacc tggacaccct gtggaaactg aacgccacca tgttcagcaa
     6181 cagcaccaac gacattctga tcatggaact gaagaacgcc agcctccagg accagggcga
     6241 ttatgtgtgt ctggcccagg acagaaagac caaaaagcgg cactgtgtcg tgcggcagct
     6301 gaccgtgctt gaaagagtgg ccccaaccat caccggcaac ctggaaaatc agaccaccag
     6361 catcggcgag tccatcgagg tgtcatgtac cgcctctggc aacccacctc cacagatcat
     6421 gtggttcaag gacaacgaga cactggtcga ggacagcggc atcgtgctga aggacggcaa
     6481 tcggaatctg accattcgga gagtgcggaa agaggacgag ggactgtata cctgccaggc
     6541 ctgtagcgtg ctgggctgtg ctaaagtgga agccttcttt atcatcgagg gcgctcaaga
     6601 aaagaccaac ctcgaaatca tcatcctcgt gggcacagcc gtgatcgcca tgttcttttg
     6661 gctgctgctc gtgatcatcc tgcggaccgt gaaaggcgcc gagagcttgt ttaaggggcc
     6721 gcgtgattac aacccgatat cgagcaccat ttgtcatttg acgaatgaat ctgatgggca
     6781 cacaacatcg ttgtatggta ttggatttgg tcccttcatc attacaaaca agcacttgtt
     6841 tagaagaaat aatggaacac tgttggtcca atcactacat ggtgtattca aggtcaagaa
     6901 caccacgact ttgcaacaac acctcattga tgggagggac atgataatta ttcgcatgcc
     6961 taaggatttc ccaccatttc ctcaaaagct gaaatttaga gagccacaaa gggaagagcg
     7021 catatgtctt gtgacaacca acttccaaac ttaggcggcc gctcgagtct agagggcccg
     7081 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     7141 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     7201 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     7261 ggcaggacag caagggggag gattgggaag tcaatagcag gcatgctggg gatgcggtgg
     7321 gctctatggc ttctgaggca ggtcgaagaa ctatttcctg gctgttacgc gagcgcatcg
     7381 cgataagctc tagagggaca gccccccccc aaagccccca gggatgtaat tacgtccctc
     7441 ccccgctagg gggcagcagc gagccgcccg gggctccgct ccggtccggc gctccccccg
     7501 catccccgag ccggcagcgt gcggggacag cccgggcacg gggaaggtgg cacgggatcg
     7561 ctttcctctg aacgcttctc gctgctcttt gagcctgcag acacctgggg ggatacgggg
     7621 aaaaagcttt aggctgaaag agagatttag aatgacagaa tcatagaacg gcctgcgtac
     7681 ggatggggct ctagctctag agcttgatat cgaattcctg cagccccgct agagggacag
     7741 ccccccccca aagcccccag ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg
     7801 agccgcccgg ggctccgctc cggtccggcg ctccccccgc atccccgagc cggcagcgtg
     7861 cggggacagc ccgggcacgg ggaaggtggc acgggatcgc tttcctctga acgcttctcg
     7921 ctgctctttg agcctgcaga cacctggggg gatacgggga aaaagcttta ggctgaaaga
     7981 gagatttaga atgacagaat catagaacgg cctgggcgcg cctggtggcg aacgatcaac
     8041 agacgggaga ggactaaata gtagttcaga cgccgttaag cgcaagatct ggggaggtcg
     8101 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     8161 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
     8221 cgcgtgagta attcatacaa aaggactcgc ccctgccttg gggaatccca gggaccgtcg
     8281 ttaaactccc actaacgtag aacccagaga tcgctgcgtt cccgccccct cacccgcccg
     8341 ctctcgtcat cactgaggtg gagaagagca tgcgtgaggc tccggtgccc gtcagtgggc
     8401 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg
     8461 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     8521 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     8581 tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg
     8641 cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacgcccc tggctgcagt
     8701 acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg
     8761 cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcttgggcgc tggggccgcc
     8821 gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca
     8881 tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg
     8941 cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc
     9001 gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg
     9061 gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta
     9121 tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat
     9181 ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc
     9241 gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat
     9301 gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga
     9361 gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt
     9421 gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc
     9481 tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc
     9541 ttccatttca ggtgtcgtga gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
     9601 tccacgctgt tttgacctcc atagaagaga acgggaccga tccaaattct cgaggctagc
     9661 tgtacagcca ccatggtgtc ctactgggat accggcgttc tgctgtgtgc cctgctgtct
     9721 tgtctgctgc tgaccggatc tagcagcggc gactacaagg accacgacgg cgattataag
     9781 gatcatgaca tcgattacaa ggacgatgac gacaagagca agctgaagga ccccgagctg
     9841 agcctgaagg gaacccagca tattatgcag gccggccaga cactgcacct ccagtgtaga
     9901 ggcgaagccg ctcacaaatg gtccctgcct gagatggtgt ccaaagagag cgagcggctg
     9961 agcatcacca agagcgcctg tggcagaaac ggcaagcagt tctgctccac actgaccctg
    10021 aatactgccc aggccaacca caccggcttc tacagctgca agtatctggc cgtgcctacc
    10081 agcaagaaga aagaaaccga gagcgccatc tacatcttca tcagcgacac cggcagaccc
    10141 ttcgtggaaa tgtactctga gatccccgag atcatccaca tgaccgaggg cagagagctg
    10201 gtcatccctt gcagagtgac aagccccaac atcaccgtga ctctgaagaa gttccctctg
    10261 gacacactga tccccgacgg caagagaatc atctgggaca gccggaaggg cttcatcatc
    10321 agcaacgcca cctacaaaga gatcggcctg ctgacctgtg aagccaccgt gaacggccac
    10381 ctgtataaga ccaactacct gacacacagg cagaccaaca ccatcatcga cgtgcagatc
    10441 agcacccctc ggccagtgaa actgctgaga ggacacaccc tggtgctgaa ttgcaccgcc
    10501 accacacctc tgaacaccag agtgcagatg acctggtctt accccgacga gaagaacaag
    10561 cgggcttccg tgcggagaag aatcgaccag agcaacagcc acgccaacat cttctactcc
    10621 gtgctgacca tcgacaagat gcagaacaag gacaagggcc tgtatacctg tagagtgcgg
    10681 agcggcccta gcttcaagag cgtgaacacc tccgtgcata tctacgacaa ggccttcatc
    10741 acagtgaagc accggaagca gcaggtcctg gaaacagtgg ccggcaagag gtcctacaga
    10801 ctgagcatga aagtgaaggc tttcccatct cctgaggtcg tgtggctgaa ggatggactg
    10861 cctgccacag agaagtctgc cagatacctg accagaggct acagcctgat catcaaggac
    10921 gtgaccgagg aagatgccgg caactacacc atcctgctgt ccatcaagca gagcaacgtg
    10981 ttcaagaacc tgaccgccac tctgatcgtg aacgtgaagc cccaaatcta tgagaaggcc
    11041 gtgtccagct ttcccgatcc tgctctgtat cccctgggct ccagacagat cctgacctgt
    11101 accgcctatg gcatccctca gcctaccatc aagtggttct ggcacccctg caaccacaac
    11161 cacagcgagg ccagatgcga cttctgtagc aacaacgagg aatccttcat cctggacgcc
    11221 gacagcaaca tgggcaacag aatcgagagc atcacacagc gcatggccat cattgagggc
    11281 aagaacaaga tggcctcaac tctggtggtg gccgactcca gaatcagcgg catctatatc
    11341 tgtatcgcca gcaacaaagt gggcaccgtg ggcagaaaca tcagcttcta catcaccgat
    11401 gtgcccaacg gcttccacgt gaacctggaa aagatgccca ccgaaggcga ggacctgaag
    11461 ctgagctgta ccgtgaacaa gttcctgtac cgggatgtga cctggattct gctgcggaca
    11521 gtgaacaacc ggaccatgca ctacagcatc agcaagcaga agatggccat taccaaagag
    11581 cacagcatca ccctgaatct gaccatcatg aacgtgtccc tgcaagacag cggcacctac
    11641 gcctgtagag cccggaatgt gtataccggc gaggaaatcc tgcaaaagaa agagatcacc
    11701 atccgggacc aagaggcccc ttacctgctg agaaacctga gcgatcacac cgtggccatc
    11761 agctccagca ccacactgga ctgtcatgcc aatggcgtgc ccgagcctca gatcacctgg
    11821 tttaagaaca accacaagat ccagcaagag cccggcatca ttctcggccc tggaagctct
    11881 accctgttca tcgagagagt gaccgaagag gacgagggcg tgtaccactg caaggccacc
    11941 aatcagaaag gcagcgtgga aagcagcgcc tacctgaccg ttcagggcac cagcgacaag
    12001 tccaacctgg aactgatcac tctgacctgc acctgtgtgg ccgctacact gttttggctg
    12061 ctgctcaccc tgtttattcg gaagatgaag cggggcgcca agagcatgtc tagcatggtg
    12121 tcagacacta gttgcacatt cccttcatct gatggcatat tctggaagca ttggattcaa
    12181 accaaggatg ggcagtgtgg cagtccatta gtatcaacta gagatgggtt cattgttggt
    12241 atacactcag catcgaattt caccaacaca aacaattatt tcacaagcgt gccgaaaaac
    12301 ttcatggaat tgaaaacaaa tcaggaggcg cagcagtggg ttagtggttg gcgattaaat
    12361 gctgactcag tattgtgggg gggccataaa gttttcatgg tgaaacctga agagcctttt
    12421 cagccagtta aggaagcgac tcaactcatg aatcttaagg gctcggagaa cctgtatttt
    12481 cagatggggg gaggcagcgg gggccctaag aaaaagcgca aagtctccgg agacgctctg
    12541 gatgactttg atctggacat gcttggttcc gatgccttgg acgattttga cctcgatatg
    12601 ctgggctccg acgctctcga cgatttcgac ctcgacatgc tgggtagcga tgcactggat
    12661 gactttgatc tggatatgct cggatccggt ggtggcgggt ctggaggagg aggtagtggt
    12721 ggaggtggct ctggtaccgc tagacccggc gaaagacctt tccagtgccg gatctgcatg
    12781 aggaacttca gcaagggcga gagactcgtg cggcacacca gaacacacac aggcgagaag
    12841 cccttccagt gtagaatctg tatgcgcaac ttcagccgga tggacaacct gagcacccac
    12901 ctgagaaccc ataccgggga gaagccattt caatgccgca tctgtatgag aaatttttcc
    12961 cggaaggacg ccctgaaccg gcacctgaaa acacacctga gaggcagctg agcggccgct
    13021 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
    13081 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
    13141 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
    13201 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagtca atagcaggca
    13261 tgctggggat gcggtgggct ctatggcttc tgaggcaggt cgaagagact gcccgacaca
    13321 tcttagtgag cgcatcgcga taagctctag agggacagcc cccccccaaa gcccccaggg
    13381 atgtaattac gtccctcccc cgctaggggg cagcagcgag ccgcccgggg ctccgctccg
    13441 gtccggcgct ccccccgcat ccccgagccg gcagcgtgcg gggacagccc gggcacgggg
    13501 aaggtggcac gggatcgctt tcctctgaac gcttctcgct gctctttgag cctgcagaca
    13561 cctgggggga tacggggaaa aagctttagg ctgaaagaga gatttagaat gacagaatca
    13621 tagaacggcc tgcgtacgga tggggctcta gctctagagc ttgatatcga attcctgcag
    13681 ccccgctaga gggacagccc ccccccaaag cccccaggga tgtaattacg tccctccccc
    13741 gctagggggc agcagcgagc cgcccggggc tccgctccgg tccggcgctc cccccgcatc
    13801 cccgagccgg cagcgtgcgg ggacagcccg ggcacgggga aggtggcacg ggatcgcttt
    13861 cctctgaacg cttctcgctg ctctttgagc ctgcagacac ctggggggat acggggaaaa
    13921 agctttaggc tgaaagagag atttagaatg acagaatcat agaacggcct gggcgcgcct
    13981 ggtggcgaac gatcaacaga cgggagaggt tacaatatcc cgtgaagctt gagtggaatc
    14041 aagatctggg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
    14101 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
    14161 tgtacgggcc agatatacgc gtgagtaatt catacaaaag gactcgcccc tgccttgggg
    14221 aatcccaggg accgtcgtta aactcccact aacgtagaac ccagagatcg ctgcgttccc
    14281 gccccctcac ccgcccgctc tcgtcatcac tgaggtggag aagagcatgc gtgaggctcc
    14341 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
    14401 gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
    14461 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
    14521 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt
    14581 ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc
    14641 acgcccctgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa gtgggtggga
    14701 gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg aggcctggct
    14761 tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt
    14821 cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt ttttctggca
    14881 agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt ttggggccgc
    14941 gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc
    15001 gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg
    15061 cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt
    15121 tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa aatggaggac
    15181 gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg cctttccgtc
    15241 ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta
    15301 gttctcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga
    15361 gtttccccac actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt
    15421 ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt
    15481 ggttcaaagt ttttttcttc catttcaggt gtcgtgagag ctcgtttagt gaaccgtcag
    15541 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagagaacg ggaccgatcc
    15601 aaattctcga ggctagcgcc accatggttt ccaagggcga agaggacaat atggccagcc
    15661 tgcctgccac acacgagctg cacatctttg gcagcatcaa cggcgtggac ttcgacatgg
    15721 tcggacaagg caccggcaat cccaacgacg gctacgagga actgaacctg aagtccacaa
    15781 agggcgacct ccagttcagc ccctggattc tggtgcctca catcggctac ggcttccacc
    15841 agtacctgcc ttatcctgac ggcatgagcc ctttccaggc cgccatggtt gatggctctg
    15901 gctaccaggt ccaccggacc atgcagtttg aggatggcgc ttccctgacc gtgaactaca
    15961 gatacaccta cgagggcagc cacatcaaag gcgaggccca agtgaaaggc acaggcttcc
    16021 ctgctgatgg ccccgtgatg accaattctc tgacagccgc cgactggtgc agaagcaaga
    16081 aaacataccc caacgacaag accatcatca gcaccttcaa gtggtcctac accaccggca
    16141 acggcaagcg gtacagaagc accgccagaa ccacctacac cttcgccaaa cctatggccg
    16201 ccaactacct gaagaaccag cctatgtacg tgttccgcaa gaccgagctg aagcactcca
    16261 agacagaact gaacttcaaa gagtggcaga aagccttcac cgacgtgatg ggcatggacg
    16321 aactctacaa agccacgaac ttcagcctgc tgaagcaggc tggagacgtg gaggagaacc
    16381 ctggacccat ggccaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
    16441 cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
    16501 tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
    16561 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
    16621 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggctcg cgcatggccg
    16681 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
    16741 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
    16801 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gctggggtgc
    16861 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
    16921 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
    16981 ccggtgcctg agcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
    17041 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
    17101 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
    17161 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
    17221 tgggaagtca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcaggt
    17281 cgacactgtt taacaagccc gtcagtagag cgcatcgcga taagctctag agggacagcc
    17341 cccccccaaa gcccccaggg atgtaattac gtccctcccc cgctaggggg cagcagcgag
    17401 ccgcccgggg ctccgctccg gtccggcgct ccccccgcat ccccgagccg gcagcgtgcg
    17461 gggacagccc gggcacgggg aaggtggcac gggatcgctt tcctctgaac gcttctcgct
    17521 gctctttgag cctgcagaca cctgggggga tacggggaaa aagctttagg ctgaaagaga
    17581 gatttagaat gacagaatca tagaacggcc tgcgtacgga tggggctcta gctctagagc
    17641 ttgatatcga attcctgcag ccccgctaga gggacagccc ccccccaaag cccccaggga
    17701 tgtaattacg tccctccccc gctagggggc agcagcgagc cgcccggggc tccgctccgg
    17761 tccggcgctc cccccgcatc cccgagccgg cagcgtgcgg ggacagcccg ggcacgggga
    17821 aggtggcacg ggatcgcttt cctctgaacg cttctcgctg ctctttgagc ctgcagacac
    17881 ctggggggat acggggaaaa agctttaggc tgaaagagag atttagaatg acagaatcat
    17941 agaacggcct gggcgcgcct ggtggcgaac gatcaacaga cgggagaggc agagaggatg
    18001 cacatgtgac cgagggatcc agatgctagt taaaagtttt gttactttat agaagaaatt
    18061 ttgagttttt gttttttttt aataaataaa taaacataaa taaattgttt gttgaattta
    18121 ttattagtat gtaagtgtaa atataataaa acttaatatc tattcaaatt aataaataaa
    18181 cctcgatata cagaccgata aaacacatgc gtcaatttta cgcatgatta tctttaacgt
    18241 acgtcacaat atgattatct ttctagggtt aatctagtat acgcgtgagc aaaaggtctg
    18301 acgagaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
    18361 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
    18421 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
    18481 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
    18541 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
    18601 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
    18661 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
    18721 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
    18781 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
    18841 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
    18901 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
    18961 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
    19021 ctttttacgg ttcctggcct tttgctggag ctagcatggc ccgtgtctca aaatctctga
    19081 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
    19141 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
    19201 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
    19261 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
    19321 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
    19381 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
    19441 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
    19501 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
    19561 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
    19621 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
    19681 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
    19741 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
    19801 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
    19861 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
    19921 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
    19981 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
    20041 tgacggtcgc caccgctaga ttaaccctag aaagatagtc tgcgtaaaat tgacgcatgc
    20101 attcttgaaa tattgctctc tctttctaaa tagcgcgaat ccgtcgctgt gcatttagga
    20161 catctcagtc gccgcttgga gctcccgtga ggcgtgcttg tcaatgcggt aagtgtcact
    20221 gattttgaac tataacgacc gcgtgagtca aaatgacgca tgattatctt ttacgtgact
    20281 tttaagattt aactcatacg ataattatat tgttatttca tgttctactt acgtgataac
    20341 ttattatata tatattttct tgttatagat atcattgatg agtttggaca aacc
//
