LOCUS       sequence-380399-       15403 bp    DNA     circular SYN 14-APR-2025
DEFINITION  FlpO-based digitizer circuit driven by AP1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15403)
  AUTHORS   Wong NM, Frias E, Sigoillot FD, Letendre JH, Hild M, Wong WW
  TITLE     Engineering digitizer circuits for chemical and genetic screens in 
            human cells.
  JOURNAL   Nat Commun. 2021 Oct 22;12(1):6150. doi: 10.1038/s41467-021-26359-9.
  PUBMED    34686672
REFERENCE   2  (bases 1 to 15403)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2021-10-22"; volume: "12(1):6150. doi"; pages: " 
            10.1038/s41467-021-26359-9"
COMMENT     Sequence Label: 
            pNW538_pAAVS1-pAP1(2)-FlpO-ABI-P2A-PYL-FlpO-CAG-U1-frt-STOP-frt-U2-G
            FP-P2A-luc2-ZeoR
FEATURES             Location/Qualifiers
     source          1..15403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    127..656
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        938..1000
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     CDS             3183..3206
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3207..3227
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3237..3293
                     /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             3834..3860
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3984..4004
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4014..4238
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        4415..4718
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          4995..6011
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     5935..5957
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     6019..6038
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    6129..6162
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             6205..6999
                     /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(6259..6278)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6869..6888
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7179..7313
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    7322..7355
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     regulatory      7405..7414
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7411..7413,7414..7416,7417..8127)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7411..7413,7414..7416,7417..8127)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7456..7477)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7717..7736)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8064..8085
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             8137..8193
                     /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             8194..9846
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(8260..8278)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    9871..10095
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        10160..10659
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(10195..10218)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     10592..10611
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      10677..10686
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             11064..11120
                     /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             11121..11822
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    11941..12062
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    12203..12704
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(12906..12922)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12906..12922)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12919..12941)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12930..12946
                     /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(12954..12960,12961..12978,12979..12984))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12999..13020
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        13079..13274
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      13125..13260
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    13280..13414
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13482..13499)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13653..14241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13733..13752)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14412..15203,15204..15272))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     15035..15054
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15273..15377)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agtgaagggg caggagagcc aggggcatga gatggtggac
       61 gaggaagggg gacagggaag cctgagcgcc tctcctgggc ttgccaagga ctcaaaccca
      121 gaagcccaga gcagggcctt agggaagcgg gaccctgctc tgggcggagg aatatgtccc
      181 agatagcact ggggactctt taaggaaaga aggatggaga aagagaaagg gagtagaggc
      241 ggccacgacc tggtgaacac ctaggacgca ccattctcac aaagggagtt ttccacacgg
      301 acacccccct cctcaccaca gccctgccag gacggggctg gctactggcc ttatctcaca
      361 ggtaaaactg acgcacggag gaacaatata aattggggac tagaaaggtg aagagccaaa
      421 gttagaactc aggaccaact tattctgatt ttgtttttcc aaactgcttc tcctcttggg
      481 aagtgtaagg aagctgcagc accaggatca gtgaaacgca ccagacagcc gcgtcagagc
      541 agctcaggtt ctgggagagg gtagcgcagg gtggccactg agaaccgggc aggtcacgca
      601 tccccccctt ccctcccacc ccctgccaag ctctccctcc caggatcctc tctggctcca
      661 tcgtaagcaa accttagagg ttctggcaag gagagagatg gctccaggaa atgggggtgt
      721 gtcaccagat aaggaatctg cctaacagga ggtgggggtt agacccaata tcaggagact
      781 aggaaggagg aggcctaagg atggggcttt tctgtcacca atcgccggcg aattcgcggc
      841 cgccttacgc gtgctagctg actaatgact aatgactaat gactaatgac taatgactaa
      901 tgactaatga ctaaagatct gggtatatag tttaaacttc gcatattaag gtgacgcgtg
      961 tggcctcgaa caccgagcga ccctgcagcg acccgcttaa gatccattac tcgcatccat
     1021 tctcaggctg tctcgtctcg tctcggccgg ccgccgccac catgagccag ttcgacatcc
     1081 tgtgcaagac cccccccaag gtgctggtgc ggcagttcgt ggagagattc gagaggccca
     1141 gcggcgagaa gatcgccagc tgtgccgccg agctgaccta cctgtgctgg atgatcaccc
     1201 acaacggcac cgccatcaag agggccacct tcatgagcta caacaccatc atcagcaaca
     1261 gcctgagctt cgacatcgtg aacaagagcc tgcagttcaa gtacaagacc cagaaggcca
     1321 ccatcctgga ggccagcctg aagaagctga tccccgcctg ggagttcacc atcatccctt
     1381 acaacggcca gaagcaccag agcgacatca ccgacatcgt gtccagcctg cagctgcagt
     1441 tcgagagcag cgaggaggcc gacaagggca acagccacag caagaagatg ctgaaggccc
     1501 tgctgtccga gggcgagagc atctgggaga tcaccgagaa gatcctgaac agcttcgagt
     1561 acaccagcag gttcaccaag accaagaccc tgtaccagtt cctgttcctg gccacattca
     1621 tcaactgcgg caggttcagc gacatcaaga acgtggaccc caagagcttc aagctggtgc
     1681 agaacaagta cctgggcgtg atcattcagt gcctggtgac cgagaccaag acaagcgtgt
     1741 ccaggcacat ctactttttc agcgccagag gcaggatcga ccccctggtg tacctggacg
     1801 agttcctgag gaacagcgag cccgtgctga agagagtgaa caggaccggc aacagcagca
     1861 gcaacaagca ggagtaccag ctgctgaagg acaacctggt gcgcagctac aacaaggccc
     1921 tgaagaagaa cgccccctac cccatcttcg ctatcaagaa cggccctaag agccacatcg
     1981 gcaggcacct gatgaccagc tttctgagca tgaagggcct gaccgagctg acaaacgtgg
     2041 tgggcaactg gagcgacaag agggcctccg ccgtggccag gaccacctac acccaccaga
     2101 tcaccgccat ccccgaccac tacttcgccc tggtgtccag gtactacgcc tacgacccca
     2161 tcagcaagga gatgatcgcc ctgaaggacg agaccaaccc catcgaggag tggcagcaca
     2221 tcgagcagct gaagggcagc gccgagggct ccggagggtc tggctccgga tcaagtggtg
     2281 gcagcggtac ccctttgtat ggttttactt cgatttgtgg aagaagacct gagatggaag
     2341 atgctgtttc gactatacca agattccttc aatcttcctc tggttcgatg ttagatggtc
     2401 ggtttgatcc tcaatccgcc gctcatttct tcggtgttta cgacggccat ggcggttctc
     2461 aggtagcgaa ctattgtaga gagaggatgc atttggcttt ggcggaggag atagctaagg
     2521 agaaaccgat gctctgcgat ggtgatacgt ggctggagaa gtggaagaaa gctcttttca
     2581 actcgttcct gagagttgac tcggagattg ggtcagttgc gccggagacg gttgggtcaa
     2641 cgtcggtggt tgccgttgtt ttcccgtctc acatcttcgt cgctaactgc ggtgactcta
     2701 gagccgttct ttgccgcggc aaaactgcac ttccattatc cgttgaccat aaaccggata
     2761 gagaagatga agctgcgagg attgaagccg caggagggaa agtgattcag tggaatggag
     2821 ctcgtgtttt cggtgttctc gccatgtcga gatccattgg cgatagatac ttgaaaccat
     2881 ccatcattcc tgatccggaa gtgacggctg tgaagagagt aaaagaagat gattgtctga
     2941 ttttggcgag tgacggggtt tgggatgtaa tgacggatga agaagcgtgt gagatggcaa
     3001 ggaagcggat tctcttgtgg cacaagaaaa acgcggtggc tggggatgca tcgttgctcg
     3061 cggatgagcg gagaaaggaa gggaaagatc ctgcggcgat gtccgcggct gagtatttgt
     3121 caaagctggc gatacagaga ggaagcaaag acaacataag tgtggtggtg gttgatttga
     3181 aggattacaa ggacgatgac gataagccca agaaaaagcg gaaggtggga agcggagcta
     3241 ctaacttcag cctgctgaag caggctggag acgtggagga gaaccctgga cctatggcgc
     3301 caactcaaga cgagttcacc caactctccc aatcaatcgc cgagttccac acgtaccaac
     3361 tcggtaacgg ccgttgctca tctctcctag ctcagcgaat ccacgcgccg ccggaaacag
     3421 tatggtccgt ggtgagacgt ttcgataggc cacagattta caaacacttc atcaaaagct
     3481 gtaacgtgag tgaagatttc gagatgcgag tgggatgcac gcgcgacgtg aacgtgataa
     3541 gtggattacc ggcgaatacg tctcgagaga gattagatct gttggacgat gatcggagag
     3601 tgactgggtt tagtataacc ggtggtgaac ataggctgag gaattataaa tcggttacga
     3661 cggttcatag atttgagaaa gaagaagaag aagaaaggat ctggaccgtt gttttggaat
     3721 cttatgttgt tgatgtaccg gaaggtaatt cggaggaaga tacgagattg tttgctgata
     3781 cggttattag attgaatctt cagaaacttg cttcgatcac tgaagctatg aactacccat
     3841 acgatgttcc agattacgct tccggagggt ctggctccgg atcaagtggt ggcagcggta
     3901 ccagcatcag ataccccgcc tggaacggca tcatcagcca ggaggtgctg gactacctga
     3961 gcagctacat caacaggcgg atccccaaga aaaagcggaa ggtgtgagaa ttcctgtgcc
     4021 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     4081 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     4141 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4201 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     4261 ctggggctct agggggtatc cccacgcgcc ctgtagcggt atacgctggg agttcgtaga
     4321 cggaaacaaa cgcagaatcc aagcgcccgg gcactagtta ttaatagtaa tcaattacgg
     4381 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     4441 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     4501 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     4561 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     4621 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     4681 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct
     4741 gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt
     4801 aattattttg tgcagcgatg ggggcggggg ggggggggcg cgcgccaggc ggggcggggs
     4861 ggggsggggg ggggggsggg gsgrggsggr grggkgsggc ggcagccaat cagagcggcg
     4921 cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa
     4981 gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg
     5041 cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg
     5101 acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc
     5161 tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct
     5221 cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg
     5281 gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg
     5341 ggagcgcggc cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct
     5401 gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc
     5461 aaccccccct gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc
     5521 tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg
     5581 tgccgggcgg ggcggggccg cctcgggccg gggrgggctc gggggrgggg cgcggcggcc
     5641 cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat
     5701 cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga
     5761 ggcgccgccg caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg
     5821 aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc
     5881 ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg
     5941 gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct
     6001 ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag
     6061 aattgattaa ttcgagcgaa gcactgaagg tcctcaatcg cactggaaac atcaaggtcg
     6121 atttaaatga agttcctatt ctctagaaag tataggaact tccggatttg atctgatcaa
     6181 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     6241 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     6301 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     6361 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     6421 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     6481 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     6541 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     6601 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     6661 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     6721 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     6781 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     6841 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     6901 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     6961 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     7021 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     7081 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     7141 gggatctcat gctggagttc ttcgcccacc ccatcgataa cttgtttatt gcagcttata
     7201 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     7261 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaaatccg
     7321 agaagttcct attctctaga aagtatagga acttctcgcg actgacctcc tgccagcaat
     7381 agtaagacaa cacgcaaagt cgccgccacc atggtgagca agggcgagga gctgttcacc
     7441 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     7501 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     7561 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     7621 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     7681 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     7741 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     7801 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     7861 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     7921 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     7981 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     8041 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     8101 actctcggca tggacgagct gtacaaggga agcggagcta ctaacttcag cctgctgaag
     8161 caggctggag acgtggagga gaaccctgga cctatggaag acgccaaaaa cataaagaaa
     8221 ggcccggcgc cattctatcc gctggaagat ggaaccgctg gagagcaact gcataaggct
     8281 atgaagagat acgccctggt tcctggaaca attgctttta cagatgcaca tatcgaggtg
     8341 gacatcactt acgctgagta cttcgaaatg tccgttcggt tggcagaagc tatgaaacga
     8401 tatgggctga atacaaatca cagaatcgtc gtatgcagtg aaaactctct tcaattcttt
     8461 atgccggtgt tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa cgacatttat
     8521 aatgaacgtg aattgctcaa cagtatgggc atttcgcagc ctaccgtggt gttcgtttcc
     8581 aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaagc tcccaatcat ccaaaaaatt
     8641 attatcatgg attctaaaac ggattaccag ggatttcagt cgatgtacac gttcgtcaca
     8701 tctcatctac ctcccggttt taatgaatac gattttgtgc cagagtcctt cgatagggac
     8761 aagacaattg cactgatcat gaactcctct ggatctactg gtctgcctaa aggtgtcgct
     8821 ctgcctcata gaactgcctg cgtgagattc tcgcatgcca gagatcctat ttttggcaat
     8881 caaatcattc cggatactgc gattttaagt gttgttccat tccatcacgg ttttggaatg
     8941 tttactacac tcggatattt gatatgtgga tttcgagtcg tcttaatgta tagatttgaa
     9001 gaagagctgt ttctgaggag ccttcaggat tacaagattc aaagtgcgct gctggtgcca
     9061 accctattct ccttcttcgc caaaagcact ctgattgaca aatacgattt atctaattta
     9121 cacgaaattg cttctggtgg cgctcccctc tctaaggaag tcggggaagc ggttgccaag
     9181 aggttccatc tgccaggtat caggcaagga tatgggctca ctgagactac atcagctatt
     9241 ctgattacac ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt
     9301 gaagcgaagg ttgtggatct ggataccggg aaaacgctgg gcgttaatca aagaggcgaa
     9361 ctgtgtgtga gaggtcctat gattatgtcc ggttatgtaa acaatccgga agcgaccaac
     9421 gccttgattg acaaggatgg atggctacat tctggagaca tagcttactg ggacgaagac
     9481 gaacacttct tcatcgttga ccgcctgaag tctctgatta agtacaaagg ctatcaggtg
     9541 gctcccgctg aattggaatc catcttgctc caacacccca acatcttcga cgcaggtgtc
     9601 gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt tttggagcac
     9661 ggaaagacga tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg
     9721 aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa
     9781 ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca agaagggcgg aaagatcgcc
     9841 gtgtaagata ctgatccctg caggggtacc ctgtgccttc tagttgccag ccatctgttg
     9901 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     9961 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
    10021 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
    10081 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
    10141 acgcgccctg tagcgtaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc
    10201 gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat
    10261 tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct
    10321 tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg
    10381 tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa
    10441 tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg
    10501 gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg
    10561 gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct
    10621 gccgcgctgt tctcctcttc ctcatctccg ggcctttcga tatctcgcac agtgccgcca
    10681 ccatggccaa gctgacaagc gccgtgcctg tgctgaccgc cagagatgtg gctggcgccg
    10741 tggaattttg gaccgacaga ctgggcttca gccgggactt cgtggaagat gacttcgccg
    10801 gcgtcgtgcg ggacgacgtg accctgttta tctccgccgt gcaggaccag gtggtgcccg
    10861 ataatactct ggcctgggtg tgggtgcgcg gcctggatga actgtatgcc gagtggagcg
    10921 aggtggtgtc caccaacttc agggatgcca gcggccctgc catgaccgag attggagaac
    10981 agccctgggg cagagagttc gccctgagag atcctgccgg caactgcgtg cacttcgtgg
    11041 ccgaggaaca ggacggaagc ggagctacta acttcagcct gctgaagcag gctggagacg
    11101 tggaggagaa ccctggacct atgagcgagc tgattaagga gaacatgcac atgaagctgt
    11161 acatggaggg caccgtggac aaccatcact tcaagtgcac atccgagggc gaaggcaagc
    11221 cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct ctccccttcg
    11281 ccttcgacat cctggctact agcttcctct acggcagcaa gaccttcatc aaccacaccc
    11341 agggcatccc cgacttcttc aagcagtcct tccctgaggg cttcacatgg gagagagtca
    11401 ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct
    11461 gcctcatcta caacgtcaag atcagagggg tgaacttcac atccaacggc cctgtgatgc
    11521 agaagaaaac actcggctgg gaggccttca ccgagacgct gtaccccgct gacggcggcc
    11581 tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg gagccatctg atcgcaaaca
    11641 tcaagaccac atatagatcc aagaaacccg ctaagaacct caagatgcct ggcgtctact
    11701 atgtggacta cagactggaa agaatcaagg aggccaacaa cgagacctac gtcgagcagc
    11761 acgaggtggc agtggccaga tactgcgacc tccctagcaa actggggcac aagcttaatt
    11821 aaatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
    11881 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
    11941 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
    12001 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
    12061 taggcgcgcc ctgtccctag tggccccact gtggggtgga ggggacagat aaaagtaccc
    12121 agaaccagag ccacattaac cggccctggg aatataaggt ggtcccagct cggggacaca
    12181 ggatccctgg aggcagcaaa catgctgtcc tgaagtggac ataggggccc gggttggagg
    12241 aagaagacta gctgagctct cggacccctg gaagatgcca tgacaggggg ctggaagagc
    12301 tagcacagac tagagaggta aggggggtag gggagctgcc caaatgaaag gagtgagagg
    12361 tgacccgaat ccacaggaga acggggtgtc caggcaaaga aagcaagagg atggagaggt
    12421 ggctaaagcc agggagacgg ggtactttgg ggttgtccag aaaaacggtg atgatgcagg
    12481 cctacaagaa ggggaggcgg gacgcaaggg agacatccgt cggagaaggc catcctaaga
    12541 aacgagagat ggcacaggcc ccagaaggag aaggaaaagg gaacccagcg agtgaagacg
    12601 gcatggggtt gggtgaggga ggagagatgc ccggagagga cccagacacg gggaggatcc
    12661 gctcagagga catcacgtgg tgcagcgccg agaaggaagt gctccggaaa gagcatcctt
    12721 gggcagcaac acagcagaga gcaaggggaa gagggagtgg aggaagacgg aacctgaagg
    12781 aggcggcagg gaaggatctg ggccagccgt agaggtgacc caggccacaa gctgcagaca
    12841 gaaagcggca caggcccagg ggagagaatg ccctcgacct gcagcccaag cttggcgtaa
    12901 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
    12961 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
    13021 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc
    13081 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
    13141 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
    13201 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
    13261 taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa
    13321 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
    13381 caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc
    13441 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
    13501 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
    13561 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
    13621 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
    13681 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
    13741 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
    13801 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
    13861 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
    13921 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
    13981 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
    14041 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
    14101 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
    14161 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
    14221 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
    14281 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
    14341 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
    14401 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
    14461 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
    14521 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
    14581 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
    14641 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
    14701 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
    14761 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
    14821 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
    14881 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
    14941 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
    15001 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
    15061 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
    15121 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
    15181 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
    15241 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
    15301 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
    15361 caaatagggg ttccgcgcac atttccccga aaagtgccac ctg
//
