LOCUS       sequence-215872-       14873 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline inducible MLL-AF4 and GFP expression. The construct has 
            been used with CAG-rtTA for making engraftable iPSC derived HSCs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14873)
  AUTHORS   Tan YT, Ye L, Xie F, Beyer AI, Muench MO, Wang J, Chen Z, Liu H, 
            Chen SJ, Kan YW
  TITLE     Respecifying human iPSC-derived blood cells into highly engraftable 
            hematopoietic stem and progenitor cells with a single factor.
  JOURNAL   Proc Natl Acad Sci U S A. 2018 Feb 27;115(9):2180-2185. doi: 
            10.1073/pnas.1718446115. Epub 2018 Jan 31.
  PUBMED    29386396
REFERENCE   2  (bases 1 to 14873)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1073/pnas.1718446115"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2018-02-27- 27"; volume: "115"; issue: "9"; pages: 
            "2180-2185"
COMMENT     Sequence Label: pAAVS1-TRE-MA4-GFP
FEATURES             Location/Qualifiers
     source          1..14873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(894..962,963..1754)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1112..1131)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(2004..2798)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(2004..2798)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2115..2134)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2725..2744
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(3147..3449)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(3249..3268)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     3572..3594
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3586..3603
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3587..3603
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3610..3629
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3610..3628
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3669..4505
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        4522..4836
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    4530..4548
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4566..4584
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4601..4619
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4637..4655
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4673..4691
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4708..4726
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4744..4762
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4809..4833
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4851..4874
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             11808..11861
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(11862..11864,11865..11867,11868..12581)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(11862..11864,11865..11867,11868..12578)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(11907..11928)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(12168..12187)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     12515..12536
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    12708..12789
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12745..12764)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    12796..13020
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     13009..13026
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(13058..13657)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(13141..13161)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     13638..13657
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(13667..13720)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    13744..13769
                     /label=SA
                     /note="splice acceptor site"
     misc_feature    13776..14579
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        complement(14614..14632)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(14653..14669)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14653..14669)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14666..14688)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14677..14693
                     /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(14701..14707,14708..14725,14726..14731))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14746..14767
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      901 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      961 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1021 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1081 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     1141 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1201 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1261 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1321 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1381 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1441 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1501 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1561 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1621 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1681 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1741 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1801 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     1981 aggaaattgt aagcgttaat aattcagaag aactcgtcaa gaaggcgata gaaggcgatg
     2041 cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg
     2101 ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca
     2161 cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc
     2221 aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgct cgccttgagc
     2281 ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg
     2341 acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg
     2401 aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat
     2461 actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat
     2521 agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc
     2581 gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac
     2641 aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca
     2701 tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg
     2761 gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc
     2821 tcttgatcag agcttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag
     2881 tttactttgc agggcttccc aaccttacca gagggcgccc cagctggcaa ttccggttcg
     2941 cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg taagcccact gcaagctacc
     3001 tgctttctct ttgcgcttgc gttttccctt gtccagatag cccagtagct gacattcatc
     3061 cggggtcagc accgtttctg cggactggct ttctacgtga aaaggatcta ggtgaagatc
     3121 ctttttgata atctcatgcc tgacatttat attccccaga acatcaggtt aatggcgttt
     3181 ttgatgtcat tttcgcggtg gctgagatca gccacttctt ccccgataac ggagaccggc
     3241 acactggcca tatcggtggt catcatgcgc cagctttcat ccccgatatg caccaccggg
     3301 taaagttcac gggagacttt atctgacagc agacgtgcac tggccagggg gatcaccatc
     3361 cgtcgccccg gcgtgtcaat aatatcactc tgtacatcca caaacagacg ataacggctc
     3421 tctcttttat aggtgtaaac cttaaactgc cgtacgtata ggctgcgcaa ctgttgggaa
     3481 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     3541 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     3601 agtgaattgt aatacgactc actatagggc gaattgaatt tagcggccgc gaattcgccc
     3661 ttgcccttag agcagagcca ggaacccctg tagggaaggg gcaggagagc caggggcatg
     3721 agatggtgga cgaggaaggg ggacagggaa gcctgagcgc ctctcctggg cttgccaagg
     3781 actcaaaccc agaagcccag agcagggcct tagggaagcg ggaccctgct ctgggcggag
     3841 gaatatgtcc cagatagcac tggggactct ttaaggaaag aaggatggag aaagagaaag
     3901 ggagtagagg cggccacgac ctggtgaaca cctaggacgc accattctca caaagggagt
     3961 tttccacacg gacacccccc tcctcaccac agccctgcca ggacggggct ggctactggc
     4021 cttatctcac aggtaaaact gacgcacgga ggaacaatat aaattgggga ctagaaaggt
     4081 gaagagccaa agttagaact caggaccaac ttattctgat tttgtttttc caaactgctt
     4141 ctcctcttgg gaagtgtaag gaagctgcag caccaggatc agtgaaacgc accagacagc
     4201 cgcgtcagag cagctcaggt tctgggagag ggtagcgcag ggtggccact gagaaccggg
     4261 caggtcacgc atccccccct tccctcccac cccctgccaa gctctccctc ccaggatcct
     4321 ctctggctcc atcgtaagca aaccttagag gttctggcaa ggagagagat ggctccagga
     4381 aatgggggtg tgtcaccaga taaggaatct gcctaacagg aggtgggggt tagacccaat
     4441 atcaggagac taggaaggag gaggcctaag gatggggctt ttctgtcacc aatcctgtcc
     4501 ctagtaaagc ttagtactgt cgagtttact ccctatcagt gatagagaac gtatgtcgag
     4561 tttactccct atcagtgata gagaacgatg tcgagtttac tccctatcag tgatagagaa
     4621 cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt actccctatc
     4681 agtgatagag aacgtatgtc gagtttatcc ctatcagtga tagagaacgt atgtcgagtt
     4741 tactccctat cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg ggaggcctat
     4801 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gaattcgatg gactacaagg
     4861 acgacgatga caagggcaat tccgcgaaca tggcgcacag ctgtcggtgg cgcttccccg
     4921 cccgacccgg gaccaccggg ggcggcggcg gcggggggcg ccggggccta gggggcgccc
     4981 cgcggcaacg cgtcccggcc ctgctgcttc cccccgggcc cccggtcggc ggtggcggcc
     5041 ccggggcgcc cccctccccc ccggctgtgg cggccgcggc ggcggcggcg ggaagcagcg
     5101 gggctggggt tccaggggga gcggccgccg cctcagcagc ctcctcgtcg tccgcctcgt
     5161 cttcgtcttc gtcatcgtcc tcagcctctt cagggccggc cctgctccgg gtgggcccgg
     5221 gcttcgacgc ggcgctgcag gtctcggccg ccatcggcac caacctgcgc cggttccggg
     5281 ccgtgtttgg ggagagcggc gggggaggcg gcagcggaga ggatgagcaa ttcttaggtt
     5341 ttggctcaga tgaagaagtc agagtgcgaa gtcccacaag gtctccttca gttaaaacta
     5401 gtcctcgaaa acctcgtggg agacctagaa gtggctctga ccgaaattca gctatcctct
     5461 cagatccatc tgtgttttcc cctctaaata aatcagagac caaatctgga gataagatca
     5521 agaagaaaga ttctaaaagt atagaaaaga agagaggaag acctcccacc ttccctggag
     5581 taaaaatcaa aataacacat ggaaaggaca tttcagagtt accaaaggga aacaaagaag
     5641 atagcctgaa aaaaattaaa aggacacctt ctgctacgtt tcagcaagcc acaaagatta
     5701 aaaaattaag agcaggtaaa ctctctcctc tcaagtctaa gtttaagaca gggaagcttc
     5761 aaataggaag gaagggggta caaattgtac gacggagagg aaggcctcca tcaacagaaa
     5821 ggataaagac cccttcgggt ctcctcatta attctgaact ggaaaagccc cagaaagtcc
     5881 ggaaagacaa ggaaggaaca cctccactta caaaagaaga taagacagtt gtcagacaaa
     5941 gccctcgaag gattaagcca gttaggatta ttccttcttc aaaaaggaca gatgcaacca
     6001 ttgctaagca actcttacag agggcaaaaa agggggctca aaagaaaatt gaaaaagaag
     6061 cagctcagct gcagggaaga aaggtgaaga cacaggtcaa aaatattcga cagttcatca
     6121 tgcctgttgt cagtgctatc tcctcgcgga tcattaagac ccctcggcgg tttatagagg
     6181 atgaggatta tgaccctcca attaaaattg cccgattaga gtctacaccg aatagtagat
     6241 tcagtgcccc gtcctgtgga tcttctgaaa aatcaagtgc agcttctcag cactcctctc
     6301 aaatgtcttc agactcctct cgatctagta gccccagtgt tgatacctcc acagactctc
     6361 aggcttctga ggagattcag gtacttcctg aggagcggag cgatacccct gaagttcatc
     6421 ctccactgcc catttcccag tccccagaaa atgagagtaa tgataggaga agcagaaggt
     6481 attcagtgtc ggagagaagt tttggatcta gaacgacgaa aaaattatca actctacaaa
     6541 gtgcccccca gcagcagacc tcctcgtctc cacctccacc tctgctgact ccaccgccac
     6601 cactgcagcc agcctccagt atctctgacc acacaccttg gcttatgcct ccaacaatcc
     6661 ccttagcatc accatttttg cctgcttcca ctgctcctat gcaagggaag cgaaaatcta
     6721 ttttgcgaga accgacattt aggtggactt ctttaaagca ttctaggtca gagccacaat
     6781 acttttcctc agcaaagtat gccaaagaag gtcttattcg caaaccaata tttgataatt
     6841 tccgaccccc tccactaact cccgaggacg ttggctttgc atctggtttt tctgcatctg
     6901 gtaccgctgc ttcagcccga ttgttttcgc cactccattc tggaacaagg tttgatatgc
     6961 acaaaaggag ccctcttctg agagctccaa gatttactcc aagtgaggct cactctagaa
     7021 tatttgagtc tgtaaccttg cctagtaatc gaacttctgc tggaacatct tcttcaggag
     7081 tatccaatag aaaaaggaaa agaaaagtgt ttagtcctat tcgatctgaa ccaagatctc
     7141 cttctcactc catgaggaca agaagtggaa ggcttagtag ttctgagctc tcacctctca
     7201 cccccccgtc ttctgtctct tcctcgttaa gcatttctgt tagtcctctt gccactagtg
     7261 ccttaaaccc aacttttact tttccttctc attccctgac tcagtctggg gaatctgcag
     7321 agaaaaatca gagaccaagg aagcagacta gtgctccggc agagccattt tcatcaagta
     7381 gtcctactcc tctcttccct tggtttaccc caggctctca gactgaaaga gggagaaata
     7441 aagacaaggc ccccgaggag ctgtccaaag atcgagatgc tgacaagagc gtggagaagg
     7501 acaagagtag agagagagac cgggagagag aaaaggagaa taagcgggag tcaaggaaag
     7561 agaaaaggaa aaagggatca gaaattcaga gtagttctgc tttgtatcct gtgggtaggg
     7621 tttccaaaga gaaggttgtt ggtgaagatg ttgccacttc atcttctgcc aaaaaagcaa
     7681 cagggcggaa gaagtcttca tcacatgatt ctgggactga tattacttct gtgactcttg
     7741 gggatacaac agctgtcaaa accaaaatac ttataaagaa agggagagga aatctggaaa
     7801 aaaccaactt ggacctcggc ccaactgccc catccctgga gaaggagaaa accctctgcc
     7861 tttccactcc ttcatctagc actgttaaac attccacttc ctccataggc tccatgttgg
     7921 ctcaggcaga caagcttcca atgactgaca agagggttgc cagcctccta aaaaaggcca
     7981 aagctcagct ctgcaagatt gagaagagta agagtcttaa acaaaccgac cagcccaaag
     8041 cacagggtca agaaagtgac tcatcagaga cctctgtgcg aggaccccgg attaaacatg
     8101 tctgcagaag agcagctgtt gcccttggcc gaaaacgagc tgtgtttcct gatgacatgc
     8161 ccaccctgag tgccttacca tgggaagaac gagaaaagat tttgtcttcc atggggaatg
     8221 atgacaagtc atcaattgct ggctcagaag atgctgaacc tcttgctcca cccatcaaac
     8281 caattaaacc tgtcactaga aacaaggcac cccaggaacc tccagtaaag aaaggacgtc
     8341 gatcgaggcg gtgtgggcag tgtcccggct gccaggtgcc tgaggactgt ggtgtttgta
     8401 ctaattgctt agataagccc aagtttggtg gtcgcaatat aaagaagcag tgctgcaaga
     8461 tgagaaaatg tcagaatcta caatggatgc cttccaaagc ctacctgcag aagcaagcta
     8521 aagctgtgaa aaagaaagag aaaaagtcta agaccagtga aaagaaagac agcaaagaga
     8581 gcagtgttgt gaagaacgtg gtggactcta gtcagaaacc taccccatca gcaagagagg
     8641 atcctgcccc aaagaaaagc agtagtgagc ctcctccacg aaagcccgtc gaggaaaaga
     8701 gtgaagaagg gaatgtctcg gcccctgggc ctgaatccaa acaggccacc actccagctt
     8761 ccaggaagtc aagcaagcag gtctcccagc cagcactggt catcccgcct cagccaccta
     8821 ctacaggacc gccaagaaaa gaagttccca aaaccactcc tagtgagccc aagaaaaagc
     8881 agcctccacc accagaatca ggtccagagc agagcaaaca gaaaaaagtg gctccccgcc
     8941 caagtatccc tgtaaaacaa aaaccaaaag aaaaggaaaa accacctccg gtcaataagc
     9001 aggagaatgc aggcactttg aacatcctca gcactctctc caatggcaat agttctaagc
     9061 aaaaaattcc agcagatgga gtccacagga tcagagtgga ctttaaggag gattgtgaag
     9121 cagaaaatgt gtgggagatg ggaggcttag gaatcttgac ttctgttcct ataacaccca
     9181 gggtggtttg ctttctctgt gccagtagtg ggcatgtaga gtttctagag cagacctact
     9241 ccaatgaagt ccattgtgtt gaagagattc tgaaggaaat gacccattca tggccgcctc
     9301 ctttgacagc aatacatacg cctagtacag ctgagccatc caagtttcct ttccctacaa
     9361 aggactctca gcatgtcagt tctgtaaccc aaaaccaaaa acaatatgat acatcttcaa
     9421 aaactcactc aaattctcag caaggaacgt catccatgct cgaagacgac cttcagctca
     9481 gtgacagtga ggacagtgac agtgaacaaa ccccagagaa gcctccctcc tcatctgcac
     9541 ctccaagtgc tccacagtcc cttccagaac cagtggcatc agcacattcc agcagtgcag
     9601 agtcagaaag caccagtgac tcagacagtt cctcagactc agagagcgag agcagttcaa
     9661 gtgacagcga agaaaatgag cccctagaaa ccccagctcc ggagcctgag cctccaacaa
     9721 caaacaaatg gcagctggac aactggctga ccaaagtcag ccagccagct gcgccaccag
     9781 agggccccag gagcacagag cccccacggc ggcacccaga gagtaagggc agcagcgaca
     9841 gtgccacgag tcaggagcat tctgaatcca aagatcctcc ccctaaaagc tccagcaaag
     9901 ccccccgggc cccacccgaa gccccccacc ccggaaagag gagctgtcag aagtctccgg
     9961 cacagcagga gcccccacaa aggcaaaccg ttggaaccaa acaacccaaa aaacctgtca
    10021 aggcctctgc ccgggcaggt tcacggacca gcctgcaggg ggaaagggag ccagggcttc
    10081 ttccctatgg ctcccgagac cagacttcca aagacaagcc caaggtgaag acgaaaggac
    10141 ggccccgggc cgcagcaagc aacgaaccca agccagcagt gcccccctcc agtgagaaga
    10201 agaagcacaa gagctccctc cctgccccct ctaaggctct ctcaggccca gaacccgcga
    10261 aggacaatgt ggaggacagg acccctgagc actttgctct tgttcccctg actgagagcc
    10321 agggcccacc ccacagtggc agcggcagca ggactagtgg ctgccgccaa gccgtggtgg
    10381 tccaggagga cagccgcaaa gacagactcc cattgccttt gagagacacc aagctgctct
    10441 caccgctcag ggacactcct cccccacaaa gcttgatggt gaagatcacc ctagacctgc
    10501 tctctcggat accccagcct cccgggaagg ggagccgcca gaggaaagca gaagataaac
    10561 agccgcccgc agggaagaag cacagctctg agaagaggag ctcagacagc tcaagcaagt
    10621 tggccaaaaa gagaaagggt gaagcagaaa gagactgtga taacaagaaa atcagactgg
    10681 agaaggaaat caaatcacag tcatcttcat cttcatcctc ccacaaagaa tcttctaaaa
    10741 caaagccctc caggccctcc tcacagtcct caaagaagga aatgctcccc ccgccacccg
    10801 tgtcctcgtc ctcccagaag ccagccaagc ctgcacttaa gaggtcaagg cgggaagcag
    10861 acacctgtgg ccaggaccct cccaaaagtg ccagcagtac caagagcaac cacaaagact
    10921 cttccattcc caagcagaga agagtagagg ggaagggctc cagaagctcc tcggagcaca
    10981 agggttcttc cggagatact gcaaatcctt ttccagtgcc ttctttgcca aatggtaact
    11041 ctaaaccagg gaagcctcaa gtgaagtttg acaaacaaca agcagacctt cacatgaggg
    11101 aggcaaaaaa gatgaagcag aaagcagagt taatgacgga cagggttgga aaggctttta
    11161 agtacctgga agccgtcttg tccttcattg agtgcggaat tgccacagag tctgaaagcc
    11221 agtcatccaa gtcagcttac tctgtctact cagaaactgt agatctcatt aaattcataa
    11281 tgtcattaaa atccttctca gatgccacag cgccaacaca agagaaaata tttgctgttt
    11341 tatgcatgcg ttgccagtcc attttgaaca tggcgatgtt tcgttgtaaa aaagacatag
    11401 caataaagta ttctcgtact cttaataaac acttcgagag ttcttccaaa gtcgcccagg
    11461 caccttctcc atgcattgca agcacaggca caccatcccc tctttcccca atgccttctc
    11521 ctgccagctc cgtagggtcc cagtcaagtg ctggcagtgt ggggagcagt ggggtggctg
    11581 ccactatcag caccccagtc accatccaga atatgacatc ttcctatgtc accatcacat
    11641 cccatgttct taccgccttt gacctttggg aacaggccga ggccctcacg aggaagaata
    11701 aagaattctt tgctcggctc agcacaaatg tgtgcacctt ggccctcaac agcagtttgg
    11761 tggacctggt gcactataca cgacagggtt ttcagcagct acaagaagag ggcagaggat
    11821 ccctgctaac atgtggtgac gtcgaggaga atcctggccc aatggtgagc aagggcgagg
    11881 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
    11941 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
    12001 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
    12061 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
    12121 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
    12181 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
    12241 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
    12301 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
    12361 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
    12421 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
    12481 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
    12541 ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccga tatctctaga
    12601 ggatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
    12661 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
    12721 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
    12781 tttcactgcc tcgacccata gagcccaccg catccccagc atgcctgcta ttgtcttccc
    12841 aatcctcccc cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca
    12901 gacaatgcga tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca
    12961 gggtcaagga aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag
    13021 tcgaggctga tcagcgggtt taaacgggcc ctctagatca ggcaccgggc ttgcgggtca
    13081 tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc
    13141 gctcgtagaa ggggaggttg cggggcgcgg aggtctccag gaaggcgggc accccggcgc
    13201 gctcggccgc ctccactccg gggagcacga cggcgctgcc cagacccttg ccctggtggt
    13261 cgggcgagac gccgacggtg gccaggaacc acgcgggctc cttgggccgg tgcggcgcca
    13321 ggaggccttc catctgttgc tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg
    13381 ggccgatctc ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc cagaccgcca
    13441 ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc gagcccgacg cgcgtgagga
    13501 agagttcttg cagctcggtg acccgctcga tgtggcggtc cggatcgacg gtgtggcgcg
    13561 tggcggggta gtcggcgaac gcggcggcga gggtgcgtac ggccctgggg acgtcgtcgc
    13621 gggtggcgag gcgcaccgtg ggcttgtact cggtcatctc gagcctaggg ccgggattct
    13681 cctccacgtc accgcatgtt agaagacttc ctctgccctc tccgctgcca gatctctcga
    13741 ggccctgtgg gaggaagaga agaggtcaga agcttgcccc actgtggggt ggaggggaca
    13801 gataaaagta cccagaacca gagccacatt aaccggccct gggaatataa ggtggtccca
    13861 gctcggggac acaggatccc tggaggcagc aaacatgctg tcctgaagtg gacatagggg
    13921 cccgggttgg aggaagaaga ctagctgagc tctcggaccc ctggaagatg ccatgacagg
    13981 gggctggaag agctagcaca gactagagag gtaagggggg taggggagct gcccaaatga
    14041 aaggagtgag aggtgacccg aatccacagg agaacggggt gtccaggcaa agaaagcaag
    14101 aggatggaga ggtggctaaa gccagggaga cggggtactt tggggttgtc cagaaaaacg
    14161 gtgatgatgc aggcctacaa gaaggggagg cgggacgcaa gggagacatc cgtcggagaa
    14221 ggccatccta agaaacgaga gatggcacag gccccagaag gagaaggaaa agggaaccca
    14281 gcgagtgaag acggcatggg gttgggtgag ggaggagaga tgcccggaga ggacccagac
    14341 acggggagga tccgctcaga ggacatcacg tggtgcagcg ccgagaagga agtgctccgg
    14401 aaagagcatc cttgggcagc aacacagcag agagcaaggg gaagagggag tggaggaaga
    14461 cggaacctga aggaggcggc agggaaggat ctgggccagc cgtagaggtg acccaggcca
    14521 caagctgcag acagaaagcg gcacaggccc aggggagaga atgctggtca gagaaagcaa
    14581 agggcgaatt cgtttaaacc tgcaggacta gtccctttag tgagggttaa ttctgagctt
    14641 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
    14701 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
    14761 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
    14821 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gct
//
