LOCUS       sequence-489496-       14586 bp    DNA     circular SYN 16-OCT-2025
DEFINITION  Multigene vector for constitutive expression of mTagBFP2, IL-10Rb 
            NTEVp chain with human IgG VH signal peptide, and IL-10Ra CTEVp 
            chain with ZF1a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14586)
  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 14586)
  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: L4866 IL-10Rb NTEVp chain, IL-10Ra CTEVp chain...
FEATURES             Location/Qualifiers
     source          1..14586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        477..856
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        857..1060
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1011..1031
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1105..1124
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1105..1123
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1143..1152
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1913..2137
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        3186..4368
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3417..4359
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     regulatory      4475..4484
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4538..4603
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    5710..5934
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        6983..8165
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          7214..8156
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     CDS             8341..8406
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             9511..9531
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     CDS             9550..9570
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9577..9726
                     /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    10107..10331
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    11120..11344
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11390..11818
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11477..11496)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11687..11708
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11841..12037
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      11888..12023
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11950..11969
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    12157..12278
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12194..12213)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12248..12267
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12327..12343)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12327..12343)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12340..12362)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12351..12367
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(12375..12381,12382..12399,12400..12405))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12420..12441
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12558..12575)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12729..13317)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12809..12828)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(13525..13619)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(13650..14444)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(13761..13780)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     14371..14390
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
       61 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
      121 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
      181 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgccaaggt cttgacaaac
      241 gagtgctagt acaagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
      301 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      361 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      421 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      481 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      541 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      601 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      661 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      721 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      781 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      841 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      901 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      961 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     1021 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
     1081 cactgcttac tggcttatcg aaattaatac gactcactat agggagtccc aagctggcta
     1141 gcgccaccat ggtttccaag ggcgaagaac tgatcaaaga aaacatgcac atgaagctct
     1201 acatggaagg caccgtggac aaccaccact tcaagtgtac cagcgaaggc gagggcaagc
     1261 cttacgaggg cacccagacc atgagaatca aggtggtgga aggcggccct ctgcctttcg
     1321 cctttgatat cctggccacc agctttctgt acggcagcaa gaccttcatc aatcacaccc
     1381 agggcatccc cgatttcttc aagcagagct tccccgaggg cttcacctgg gagagagtga
     1441 ccacatacga ggatggcggc gtgctgacag ccacacagga tacaagcctc caggacggct
     1501 gcctgatcta caacgtgaag atccggggcg tgaacttcac cagcaacggc cccgtgatgc
     1561 agaagaaaac ccttggctgg gaagccttca ccgagacact gtatcctgcc gatggcggcc
     1621 tggaaggcag aaacgatatg gccctgaagc tcgtcggcgg ctctcacctg attgccaatg
     1681 ccaagaccac ctacagaagc aagaagcccg ccaagaacct gaagatgccc ggcgtgtact
     1741 acgtggacta ccggctggaa agaatcaaag aggccaacaa cgagacatac gtggaacagc
     1801 acgaggtggc cgtggccaga tactgtgatc tgccttctaa gctgggccac aagctgaact
     1861 gagcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     1921 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1981 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2041 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagtc
     2101 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcagg tcgagggtac
     2161 atgcgcctta ctccttgtga gcgcatcgcg ataagctcta gagggacagc ccccccccaa
     2221 agcccccagg gatgtaatta cgtccctccc ccgctagggg gcagcagcga gccgcccggg
     2281 gctccgctcc ggtccggcgc tccccccgca tccccgagcc ggcagcgtgc ggggacagcc
     2341 cgggcacggg gaaggtggca cgggatcgct ttcctctgaa cgcttctcgc tgctctttga
     2401 gcctgcagac acctgggggg atacggggaa aaagctttag gctgaaagag agatttagaa
     2461 tgacagaatc atagaacggc ctgcgtacgg atggggctct agctctagag cttgatatcg
     2521 aattcctgca gccccgctag agggacagcc cccccccaaa gcccccaggg atgtaattac
     2581 gtccctcccc cgctaggggg cagcagcgag ccgcccgggg ctccgctccg gtccggcgct
     2641 ccccccgcat ccccgagccg gcagcgtgcg gggacagccc gggcacgggg aaggtggcac
     2701 gggatcgctt tcctctgaac gcttctcgct gctctttgag cctgcagaca cctgggggga
     2761 tacggggaaa aagctttagg ctgaaagaga gatttagaat gacagaatca tagaacggcc
     2821 tgggcgcgcc tggtggcgaa cgatcaacag acgggagagg gcaaaaccga tgttgacgga
     2881 ctaatcctga caagatctgg ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     2941 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     3001 ctgcttcgcg atgtacgggc cagatatacg cgtgagtaat tcatacaaaa ggactcgccc
     3061 ctgccttggg gaatcccagg gaccgtcgtt aaactcccac taacgtagaa cccagagatc
     3121 gctgcgttcc cgccccctca cccgcccgct ctcgtcatca ctgaggtgga gaagagcatg
     3181 cgtgaggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
     3241 tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg
     3301 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
     3361 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa
     3421 gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt
     3481 gaattacttc cacgcccctg gctgcagtac gtgattcttg atcccgagct tcgggttgga
     3541 agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt
     3601 gaggcctggc ttgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt
     3661 ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt
     3721 tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt
     3781 tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg
     3841 ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct
     3901 ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg
     3961 tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca
     4021 aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg
     4081 gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg
     4141 cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt
     4201 tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac
     4261 ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag
     4321 cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgaga gctcgtttag
     4381 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagagaac
     4441 gggaccgatc caaattctcg aggctagctg tacagccacc atggagtttg gcctgagctg
     4501 gctgtttctg gtggccattc tgaagggcgt gcagtgcgac tacaaggacc acgacggcga
     4561 ttataaggat catgacatcg attacaagga cgatgacgac aagatggtgc ctccacctga
     4621 gaacgtgcgg atgaacagcg tgaacttcaa gaacatcctc cagtgggaga gccccgcctt
     4681 cgccaaggga aatctgacct tcacagccca gtacctgagc taccggatct tccaggacaa
     4741 gtgcatgaac accacactga ccgagtgcga cttcagcagc ctgtctaagt acggcgatca
     4801 caccctgaga gtgcgggccg agtttgccga tgagcacagc gattgggtca acatcacatt
     4861 ctgccccgtg gacgacacca tcatcggccc acctggaatg caggttgagg tgctggccga
     4921 tagcctgcac atgagatttc tggcccctaa gatcgagaac gagtacgaga catggaccat
     4981 gaagaacgtc tacaacagct ggacctacaa cgtgcagtac tggaagaacg gcaccgacga
     5041 gaagttccag atcacccctc agtacgactt cgaggtgctg cggaacctgg aaccttggac
     5101 cacctactgt gtgcaagtgc ggggcttcct gcctgacaga aacaaagctg gcgagtggag
     5161 cgagcctgtg tgcgagcaga caacccacga tgagacagtg ccctcttgga tggtggccgt
     5221 gatcctgatg gccagcgtgt tcatggtttg cctggctctg ctgggctgct tcgctctgct
     5281 ttggtgcgtc tacaagaagg gcgccgagag cttgtttaag gggccgcgtg attacaaccc
     5341 gatatcgagc accatttgtc atttgacgaa tgaatctgat gggcacacaa catcgttgta
     5401 tggtattgga tttggtccct tcatcattac aaacaagcac ttgtttagaa gaaataatgg
     5461 aacactgttg gtccaatcac tacatggtgt attcaaggtc aagaacacca cgactttgca
     5521 acaaagcctc attgatggga gggacatgat aattattcgc atgcctaagg atttcccacc
     5581 atttcctcaa aagctgaaat ttagagagcc acaaagggaa gagcgcatat gtcttgtgac
     5641 aaccaacttc caaacttagg cggccgctcg agtctagagg gcccgtttaa acccgctgat
     5701 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     5761 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     5821 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     5881 gggaggattg ggaagtcaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     5941 aggcaggtcg aagaactatt tcctggctgt tacgcgagcg catcgcgata agctctagag
     6001 ggacagcccc cccccaaagc ccccagggat gtaattacgt ccctcccccg ctagggggca
     6061 gcagcgagcc gcccggggct ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc
     6121 agcgtgcggg gacagcccgg gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc
     6181 ttctcgctgc tctttgagcc tgcagacacc tggggggata cggggaaaaa gctttaggct
     6241 gaaagagaga tttagaatga cagaatcata gaacggcctg cgtacggatg gggctctagc
     6301 tctagagctt gatatcgaat tcctgcagcc ccgctagagg gacagccccc ccccaaagcc
     6361 cccagggatg taattacgtc cctcccccgc tagggggcag cagcgagccg cccggggctc
     6421 cgctccggtc cggcgctccc cccgcatccc cgagccggca gcgtgcgggg acagcccggg
     6481 cacggggaag gtggcacggg atcgctttcc tctgaacgct tctcgctgct ctttgagcct
     6541 gcagacacct ggggggatac ggggaaaaag ctttaggctg aaagagagat ttagaatgac
     6601 agaatcatag aacggcctgg gcgcgcctgg tggcgaacga tcaacagacg ggagaggact
     6661 aaatagtagt tcagacgccg ttaagcgcaa gatctgggga ggtcgctgag tagtgcgcga
     6721 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     6781 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt gagtaattca
     6841 tacaaaagga ctcgcccctg ccttggggaa tcccagggac cgtcgttaaa ctcccactaa
     6901 cgtagaaccc agagatcgct gcgttcccgc cccctcaccc gcccgctctc gtcatcactg
     6961 aggtggagaa gagcatgcgt gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc
     7021 cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg
     7081 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     7141 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     7201 cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta
     7261 tggcccttgc gtgccttgaa ttacttccac gcccctggct gcagtacgtg attcttgatc
     7321 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
     7381 cgcctcgtgc ttgagttgag gcctggcttg ggcgctgggg ccgccgcgtg cgaatctggt
     7441 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
     7501 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
     7561 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
     7621 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
     7681 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
     7741 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
     7801 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
     7861 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     7921 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     7981 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     8041 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     8101 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     8161 cgtgagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
     8221 cctccataga agagaacggg accgatccaa attctcgagg ctagctgtac agccaccatg
     8281 ctgccctgcc tggttgttct gcttgccgct ctgctgtctc tgagactggg atctgacgcc
     8341 gactacaagg accacgacgg cgattataag gatcatgaca tcgattacaa ggacgatgac
     8401 gacaagcacg gcaccgagct gccttctcca cctagcgttt ggttcgaggc cgagttcttc
     8461 caccacatcc tgcactggac ccctattcct aaccagagcg agagcacctg ttacgaggtg
     8521 gccctgctga gatacggcat cgagagctgg aacagcatca gcaactgcag ccagacactg
     8581 agctacgacc tgaccgccgt gacactggat ctgtaccaca gcaacggcta ccgggccaga
     8641 gttagagccg tggatggaag cagacactcc aactggaccg tgaccaacac cagattcagc
     8701 gtggacgaag tgaccctgac agtgggcagc gtgaacctgg aaatccacaa cggcttcatc
     8761 ctgggcaaga tccagctgcc tcggcctaag atggcccctg ccaatgatac ctacgagagc
     8821 atcttcagcc acttccgcga gtacgagatc gccatcagaa aggtgcccgg caacttcacc
     8881 ttcacacaca agaaagtgaa gcacgagaac ttcagcctgc tgacctctgg cgaagtgggc
     8941 gagttctgcg tgcaagtgaa accctctgtg gccagccggt ccaacaaagg catgtggtcc
     9001 aaagaggaat gcatcagcct gaccagacag tacttcaccg tgacaaacgt gatcatcttc
     9061 ttcgccttcg tgctgctgct gtctggcgct ctggcttatt gtctggccct gcagctgtac
     9121 gtgcggcgga gaaagaaggg cgccaagagc atgtctagca tggtgtcaga cactagttgc
     9181 acattccctt catctgatgg catattctgg aagcattgga ttcaaaccaa ggatgggcag
     9241 tgtggcagtc cattagtatc aactagagat gggttcattg ttggtataca ctcagcatcg
     9301 aatttcacca acacaaacaa ttatttcaca agcgtgccga aaaacttcat ggaattgaaa
     9361 acaaatcagg aggcgcagca gtgggttagt ggttggcgat taaatgctga ctcagtattg
     9421 tgggggggcc ataaagtttt catggtgaaa cctgaagagc cttttcagcc agttaaggaa
     9481 gcgactcaac tcatgaatct taagggctcg gagaacctgt attttcagat ggggggaggc
     9541 agcgggggcc ctaagaaaaa gcgcaaagtc tccggagacg ctctggatga ctttgatctg
     9601 gacatgcttg gttccgatgc cttggacgat tttgacctcg atatgctggg ctccgacgct
     9661 ctcgacgatt tcgacctcga catgctgggt agcgatgcac tggatgactt tgatctggat
     9721 atgctcggat ccggtggtgg cgggtctgga ggaggaggta gtggtggagg tggctctggt
     9781 accgccagac ctggagaaag acctttccag tgcagaatct gtatgagaaa cttcagcaga
     9841 caggaccgcc tggacagaca caccagaaca catacaggag agaagccatt tcagtgtagg
     9901 atctgtatga ggaactttag ccagaaggag cacctggccg gacatctgcg cacacacact
     9961 ggagagaagc cattccagtg tagaatctgc atgagaaact tcagcagacg cgacaacctg
    10021 aacagacacc tcaagacaca cctgagagga agctgagcgg ccgctcgagt ctagagggcc
    10081 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
    10141 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
    10201 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
    10261 ggggcaggac agcaaggggg aggattggga agtcaatagc aggcatgctg gggatgcggt
    10321 gggctctatg gcttctgagg caggtcgaag agactgcccg acacatctta gtgagcgcat
    10381 cgcgataagc tctagaggga cagccccccc ccaaagcccc cagggatgta attacgtccc
    10441 tcccccgcta gggggcagca gcgagccgcc cggggctccg ctccggtccg gcgctccccc
    10501 cgcatccccg agccggcagc gtgcggggac agcccgggca cggggaaggt ggcacgggat
    10561 cgctttcctc tgaacgcttc tcgctgctct ttgagcctgc agacacctgg ggggatacgg
    10621 ggaaaaagct ttaggctgaa agagagattt agaatgacag aatcatagaa cggcctgcgt
    10681 acggatgggg ctctagctct agagcttgat atcgaattcc tgcagccccg ctagagggac
    10741 agcccccccc caaagccccc agggatgtaa ttacgtccct cccccgctag ggggcagcag
    10801 cgagccgccc ggggctccgc tccggtccgg cgctcccccc gcatccccga gccggcagcg
    10861 tgcggggaca gcccgggcac ggggaaggtg gcacgggatc gctttcctct gaacgcttct
    10921 cgctgctctt tgagcctgca gacacctggg gggatacggg gaaaaagctt taggctgaaa
    10981 gagagattta gaatgacaga atcatagaac ggcctgggcg cgcctggtgg cgaacgatca
    11041 acagacggga gaggttacga ggatgcacat gtgaccgagg gatctagagg gcccgtttaa
    11101 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
    11161 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
    11221 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
    11281 gacagcaagg gggaggattg ggaagtcaat agcaggcatg ctggggatgc ggtgggctct
    11341 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
    11401 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
    11461 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
    11521 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
    11581 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
    11641 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
    11701 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
    11761 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
    11821 ttctgtggaa tgtgtgtcag ttaggctcaa ttagtcagca accatagtcc cgcccctaac
    11881 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
    11941 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
    12001 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
    12061 cattttcgga ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
    12121 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
    12181 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
    12241 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
    12301 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
    12361 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
    12421 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
    12481 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
    12541 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
    12601 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
    12661 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
    12721 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
    12781 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
    12841 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
    12901 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
    12961 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
    13021 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
    13081 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
    13141 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
    13201 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
    13261 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
    13321 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
    13381 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
    13441 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgcttg
    13501 acattggtca tggctagtgc ttggattctc accaataaaa aacgcccggc ggcaaccgag
    13561 cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc
    13621 aagccaattc tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag
    13681 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
    13741 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc
    13801 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
    13861 ttcggcaagc aggcatcgcc gtgggtcacg acgagatcct cgccgtcggg catacgcgcc
    13921 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc
    13981 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg
    14041 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
    14101 atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg
    14161 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc cgcgcaagga
    14221 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cttgcagttc attcagggca
    14281 ccggacaggt cggtcttgac aaaaagaacc gggcgaccct gcgctgacag ccggaacacg
    14341 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
    14401 caagccgccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat
    14461 cctgtctctt gatcagatct tgatcccctg cgccatcaga tccttggcgg caagaaagcc
    14521 atccagttta ctttgcaggg cttcccaacc ttaccagagg gcggcccaac tggcaattcc
    14581 gacgtc
//
