LOCUS       sequence-220669-       14870 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 14870)
  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 14870)
  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..14870
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     122..144
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     136..153
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     137..153
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     160..179
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        160..178
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    213..1049
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        1066..1380
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    1074..1092
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1110..1128
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1145..1163
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1181..1199
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1217..1235
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1252..1270
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1288..1306
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1353..1377
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1395..1418
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8355..8408
                     /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(8409..8411,8412..8414,8415..9128)
                     /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(8409..8411,8412..8414,8415..9125)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8454..8475)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8715..8734)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9062..9083
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    9255..9336
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9292..9311)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    9343..9567
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     9556..9573
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(9605..10204)
                     /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(9688..9708)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     10185..10204
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(10214..10267)
                     /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    10291..10316
                     /label=SA
                     /note="splice acceptor site"
     misc_feature    10323..11126
                     /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(11161..11179)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11200..11216)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11200..11216)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11213..11235)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11224..11240
                     /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(11248..11254,11255..11272,11273..11278))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11293..11314
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11431..11448)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11602..12190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11682..11701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(12314..12382,12383..13174)
                     /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(12532..12551)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(13424..14218)
                     /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(13424..14218)
                     /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(13535..13554)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     14145..14164
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(14567..14869)
                     /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(14669..14688)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
ORIGIN
        1 cgtacgtata ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
       61 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
      121 tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc
      181 gaattgaatt tagcggccgc gaattcgccc ttagagcaga gccaggaacc cctgtaggga
      241 aggggcagga gagccagggg catgagatgg tggacgagga agggggacag ggaagcctga
      301 gcgcctctcc tgggcttgcc aaggactcaa acccagaagc ccagagcagg gccttaggga
      361 agcgggaccc tgctctgggc ggaggaatat gtcccagata gcactgggga ctctttaagg
      421 aaagaaggat ggagaaagag aaagggagta gaggcggcca cgacctggtg aacacctagg
      481 acgcaccatt ctcacaaagg gagttttcca cacggacacc cccctcctca ccacagccct
      541 gccaggacgg ggctggctac tggccttatc tcacaggtaa aactgacgca cggaggaaca
      601 atataaattg gggactagaa aggtgaagag ccaaagttag aactcaggac caacttattc
      661 tgattttgtt tttccaaact gcttctcctc ttgggaagtg taaggaagct gcagcaccag
      721 gatcagtgaa acgcaccaga cagccgcgtc agagcagctc aggttctggg agagggtagc
      781 gcagggtggc cactgagaac cgggcaggtc acgcatcccc cccttccctc ccaccccctg
      841 ccaagctctc cctcccagga tcctctctgg ctccatcgta agcaaacctt agaggttctg
      901 gcaaggagag agatggctcc aggaaatggg ggtgtgtcac cagataagga atctgcctaa
      961 caggaggtgg gggttagacc caatatcagg agactaggaa ggaggaggcc taaggatggg
     1021 gcttttctgt caccaatcct gtccctagta aagcttagta ctgtcgagtt tactccctat
     1081 cagtgataga gaacgtatgt cgagtttact ccctatcagt gatagagaac gatgtcgagt
     1141 ttactcccta tcagtgatag agaacgtatg tcgagtttac tccctatcag tgatagagaa
     1201 cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt atccctatca
     1261 gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacgt atgtcgaggt
     1321 aggcgtgtac ggtgggaggc ctatataagc agagctcgtt tagtgaaccg tcagatcgcc
     1381 tggagaattc catggactac aaggacgacg atgacaaggg caattccgcg aacatggcgc
     1441 acagctgtcg gtggcgcttc cccgcccgac ccgggaccac cgggggcggc ggcggcgggg
     1501 ggcgccgggg cctagggggc gccccgcggc aacgcgtccc ggccctgctg cttccccccg
     1561 ggcccccggt cggcggtggc ggccccgggg cgcccccctc ccccccggct gtggcggccg
     1621 cggcggcggc ggcgggaagc agcggggctg gggttccagg gggagcggcc gccgcctcag
     1681 cagcctcctc gtcgtccgcc tcgtcttcgt cttcgtcatc gtcctcagcc tcttcagggc
     1741 cggccctgct ccgggtgggc ccgggcttcg acgcggcgct gcaggtctcg gccgccatcg
     1801 gcaccaacct gcgccggttc cgggccgtgt ttggggagag cggcggggga ggcggcagcg
     1861 gagaggatga gcaattctta ggttttggct cagatgaaga agtcagagtg cgaagtccca
     1921 caaggtctcc ttcagttaaa actagtcctc gaaaacctcg tgggagacct agaagtggct
     1981 ctgaccgaaa ttcagctatc ctctcagatc catctgtgtt ttcccctcta aataaatcag
     2041 agaccaaatc tggagataag atcaagaaga aagattctaa aagtatagaa aagaagagag
     2101 gaagacctcc caccttccct ggagtaaaaa tcaaaataac acatggaaag gacatttcag
     2161 agttaccaaa gggaaacaaa gaagatagcc tgaaaaaaat taaaaggaca ccttctgcta
     2221 cgtttcagca agccacaaag attaaaaaat taagagcagg taaactctct cctctcaagt
     2281 ctaagtttaa gacagggaag cttcaaatag gaaggaaggg ggtacaaatt gtacgacgga
     2341 gaggaaggcc tccatcaaca gaaaggataa agaccccttc gggtctcctc attaattctg
     2401 aactggaaaa gccccagaaa gtccggaaag acaaggaagg aacacctcca cttacaaaag
     2461 aagataagac agttgtcaga caaagccctc gaaggattaa gccagttagg attattcctt
     2521 cttcaaaaag gacagatgca accattgcta agcaactctt acagagggca aaaaaggggg
     2581 ctcaaaagaa aattgaaaaa gaagcagctc agctgcaggg aagaaaggtg aagacacagg
     2641 tcaaaaatat tcgacagttc atcatgcctg ttgtcagtgc tatctcctcg cggatcatta
     2701 agacccctcg gcggtttata gaggatgagg attatgaccc tccaattaaa attgcccgat
     2761 tagagtctac accgaatagt agattcagtg ccccgtcctg tggatcttct gaaaaatcaa
     2821 gtgcagcttc tcagcactcc tctcaaatgt cttcagactc ctctcgatct agtagcccca
     2881 gtgttgatac ctccacagac tctcaggctt ctgaggagat tcaggtactt cctgaggagc
     2941 ggagcgatac ccctgaagtt catcctccac tgcccatttc ccagtcccca gaaaatgaga
     3001 gtaatgatag gagaagcaga aggtattcag tgtcggagag aagttttgga tctagaacga
     3061 cgaaaaaatt atcaactcta caaagtgccc cccagcagca gacctcctcg tctccacctc
     3121 cacctctgct gactccaccg ccaccactgc agccagcctc cagtatctct gaccacacac
     3181 cttggcttat gcctccaaca atccccttag catcaccatt tttgcctgct tccactgctc
     3241 ctatgcaagg gaagcgaaaa tctattttgc gagaaccgac atttaggtgg acttctttaa
     3301 agcattctag gtcagagcca caatactttt cctcagcaaa gtatgccaaa gaaggtctta
     3361 ttcgcaaacc aatatttgat aatttccgac cccctccact aactcccgag gacgttggct
     3421 ttgcatctgg tttttctgca tctggtaccg ctgcttcagc ccgattgttt tcgccactcc
     3481 attctggaac aaggtttgat atgcacaaaa ggagccctct tctgagagct ccaagattta
     3541 ctccaagtga ggctcactct agaatatttg agtctgtaac cttgcctagt aatcgaactt
     3601 ctgctggaac atcttcttca ggagtatcca atagaaaaag gaaaagaaaa gtgtttagtc
     3661 ctattcgatc tgaaccaaga tctccttctc actccatgag gacaagaagt ggaaggctta
     3721 gtagttctga gctctcacct ctcacccccc cgtcttctgt ctcttcctcg ttaagcattt
     3781 ctgttagtcc tcttgccact agtgccttaa acccaacttt tacttttcct tctcattccc
     3841 tgactcagtc tggggaatct gcagagaaaa atcagagacc aaggaagcag actagtgctc
     3901 cggcagagcc attttcatca agtagtccta ctcctctctt cccttggttt accccaggct
     3961 ctcagactga aagagggaga aataaagaca aggcccccga ggagctgtcc aaagatcgag
     4021 atgctgacaa gagcgtggag aaggacaaga gtagagagag agaccgggag agagaaaagg
     4081 agaataagcg ggagtcaagg aaagagaaaa ggaaaaaggg atcagaaatt cagagtagtt
     4141 ctgctttgta tcctgtgggt agggtttcca aagagaaggt tgttggtgaa gatgttgcca
     4201 cttcatcttc tgccaaaaaa gcaacagggc ggaagaagtc ttcatcacat gattctggga
     4261 ctgatattac ttctgtgact cttggggata caacagctgt caaaaccaaa atacttataa
     4321 agaaagggag aggaaatctg gaaaaaacca acttggacct cggcccaact gccccatccc
     4381 tggagaagga gaaaaccctc tgcctttcca ctccttcatc tagcactgtt aaacattcca
     4441 cttcctccat aggctccatg ttggctcagg cagacaagct tccaatgact gacaagaggg
     4501 ttgccagcct cctaaaaaag gccaaagctc agctctgcaa gattgagaag agtaagagtc
     4561 ttaaacaaac cgaccagccc aaagcacagg gtcaagaaag tgactcatca gagacctctg
     4621 tgcgaggacc ccggattaaa catgtctgca gaagagcagc tgttgccctt ggccgaaaac
     4681 gagctgtgtt tcctgatgac atgcccaccc tgagtgcctt accatgggaa gaacgagaaa
     4741 agattttgtc ttccatgggg aatgatgaca agtcatcaat tgctggctca gaagatgctg
     4801 aacctcttgc tccacccatc aaaccaatta aacctgtcac tagaaacaag gcaccccagg
     4861 aacctccagt aaagaaagga cgtcgatcga ggcggtgtgg gcagtgtccc ggctgccagg
     4921 tgcctgagga ctgtggtgtt tgtactaatt gcttagataa gcccaagttt ggtggtcgca
     4981 atataaagaa gcagtgctgc aagatgagaa aatgtcagaa tctacaatgg atgccttcca
     5041 aagcctacct gcagaagcaa gctaaagctg tgaaaaagaa agagaaaaag tctaagacca
     5101 gtgaaaagaa agacagcaaa gagagcagtg ttgtgaagaa cgtggtggac tctagtcaga
     5161 aacctacccc atcagcaaga gaggatcctg ccccaaagaa aagcagtagt gagcctcctc
     5221 cacgaaagcc cgtcgaggaa aagagtgaag aagggaatgt ctcggcccct gggcctgaat
     5281 ccaaacaggc caccactcca gcttccagga agtcaagcaa gcaggtctcc cagccagcac
     5341 tggtcatccc gcctcagcca cctactacag gaccgccaag aaaagaagtt cccaaaacca
     5401 ctcctagtga gcccaagaaa aagcagcctc caccaccaga atcaggtcca gagcagagca
     5461 aacagaaaaa agtggctccc cgcccaagta tccctgtaaa acaaaaacca aaagaaaagg
     5521 aaaaaccacc tccggtcaat aagcaggaga atgcaggcac tttgaacatc ctcagcactc
     5581 tctccaatgg caatagttct aagcaaaaaa ttccagcaga tggagtccac aggatcagag
     5641 tggactttaa ggaggattgt gaagcagaaa atgtgtggga gatgggaggc ttaggaatct
     5701 tgacttctgt tcctataaca cccagggtgg tttgctttct ctgtgccagt agtgggcatg
     5761 tagagtttct agagcagacc tactccaatg aagtccattg tgttgaagag attctgaagg
     5821 aaatgaccca ttcatggccg cctcctttga cagcaataca tacgcctagt acagctgagc
     5881 catccaagtt tcctttccct acaaaggact ctcagcatgt cagttctgta acccaaaacc
     5941 aaaaacaata tgatacatct tcaaaaactc actcaaattc tcagcaagga acgtcatcca
     6001 tgctcgaaga cgaccttcag ctcagtgaca gtgaggacag tgacagtgaa caaaccccag
     6061 agaagcctcc ctcctcatct gcacctccaa gtgctccaca gtcccttcca gaaccagtgg
     6121 catcagcaca ttccagcagt gcagagtcag aaagcaccag tgactcagac agttcctcag
     6181 actcagagag cgagagcagt tcaagtgaca gcgaagaaaa tgagccccta gaaaccccag
     6241 ctccggagcc tgagcctcca acaacaaaca aatggcagct ggacaactgg ctgaccaaag
     6301 tcagccagcc agctgcgcca ccagagggcc ccaggagcac agagccccca cggcggcacc
     6361 cagagagtaa gggcagcagc gacagtgcca cgagtcagga gcattctgaa tccaaagatc
     6421 ctccccctaa aagctccagc aaagcccccc gggccccacc cgaagccccc caccccggaa
     6481 agaggagctg tcagaagtct ccggcacagc aggagccccc acaaaggcaa accgttggaa
     6541 ccaaacaacc caaaaaacct gtcaaggcct ctgcccgggc aggttcacgg accagcctgc
     6601 agggggaaag ggagccaggg cttcttccct atggctcccg agaccagact tccaaagaca
     6661 agcccaaggt gaagacgaaa ggacggcccc gggccgcagc aagcaacgaa cccaagccag
     6721 cagtgccccc ctccagtgag aagaagaagc acaagagctc cctccctgcc ccctctaagg
     6781 ctctctcagg cccagaaccc gcgaaggaca atgtggagga caggacccct gagcactttg
     6841 ctcttgttcc cctgactgag agccagggcc caccccacag tggcagcggc agcaggacta
     6901 gtggctgccg ccaagccgtg gtggtccagg aggacagccg caaagacaga ctcccattgc
     6961 ctttgagaga caccaagctg ctctcaccgc tcagggacac tcctccccca caaagcttga
     7021 tggtgaagat caccctagac ctgctctctc ggatacccca gcctcccggg aaggggagcc
     7081 gccagaggaa agcagaagat aaacagccgc ccgcagggaa gaagcacagc tctgagaaga
     7141 ggagctcaga cagctcaagc aagttggcca aaaagagaaa gggtgaagca gaaagagact
     7201 gtgataacaa gaaaatcaga ctggagaagg aaatcaaatc acagtcatct tcatcttcat
     7261 cctcccacaa agaatcttct aaaacaaagc cctccaggcc ctcctcacag tcctcaaaga
     7321 aggaaatgct ccccccgcca cccgtgtcct cgtcctccca gaagccagcc aagcctgcac
     7381 ttaagaggtc aaggcgggaa gcagacacct gtggccagga ccctcccaaa agtgccagca
     7441 gtaccaagag caaccacaaa gactcttcca ttcccaagca gagaagagta gaggggaagg
     7501 gctccagaag ctcctcggag cacaagggtt cttccggaga tactgcaaat ccttttccag
     7561 tgccttcttt gccaaatggt aactctaaac cagggaagcc tcaagtgaag tttgacaaac
     7621 aacaagcaga ccttcacatg agggaggcaa aaaagatgaa gcagaaagca gagttaatga
     7681 cggacagggt tggaaaggct tttaagtacc tggaagccgt cttgtccttc attgagtgcg
     7741 gaattgccac agagtctgaa agccagtcat ccaagtcagc ttactctgtc tactcagaaa
     7801 ctgtagatct cattaaattc ataatgtcat taaaatcctt ctcagatgcc acagcgccaa
     7861 cacaagagaa aatatttgct gttttatgca tgcgttgcca gtccattttg aacatggcga
     7921 tgtttcgttg taaaaaagac atagcaataa agtattctcg tactcttaat aaacacttcg
     7981 agagttcttc caaagtcgcc caggcacctt ctccatgcat tgcaagaagc acaggcacac
     8041 catcccctct ttccccaatg ccttctcctg ccagctccgt agggtcccag tcaagtgctg
     8101 gcagtgtggg gagcagtggg gtggctgcca ctatcagcac cccagtcacc atccagaata
     8161 tgacatcttc ctatgtcacc atcacatccc atgttcttac cgcctttgac ctttgggaac
     8221 aggccgaggc cctcacgagg aagaataaag aattctttgc tcggctcagc acaaatgtgt
     8281 gcaccttggc cctcaacagc agtttggtgg acctggtgca ctatacacga cagggttttc
     8341 agcagctaca agaagagggc agaggatccc tgctaacatg tggtgacgtc gaggagaatc
     8401 ctggcccaat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     8461 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     8521 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     8581 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     8641 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     8701 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     8761 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     8821 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     8881 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     8941 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     9001 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     9061 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     9121 acaagtaaag cggccgatat ctctagagga tcataatcag ccataccaca tttgtagagg
     9181 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     9241 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     9301 tcacaaattt cacaaataaa gcattttttt cactgcctcg acccatagag cccaccgcat
     9361 ccccagcatg cctgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac
     9421 cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg
     9481 aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa
     9541 cagatggctg gcaactagaa ggcacagtcg aggctgatca gcgggtttaa acgggccctc
     9601 tagatcaggc accgggcttg cgggtcatgc accaggtgcg cggtccttcg ggcacctcga
     9661 cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg ggcgcggagg
     9721 tctccaggaa ggcgggcacc ccggcgcgct cggccgcctc cactccgggg agcacgacgg
     9781 cgctgcccag acccttgccc tggtggtcgg gcgagacgcc gacggtggcc aggaaccacg
     9841 cgggctcctt gggccggtgc ggcgccagga ggccttccat ctgttgctgc gcggccagcc
     9901 gggaaccgct caactcggcc atgcgcgggc cgatctcggc gaacaccgcc cccgcttcga
     9961 cgctctccgg cgtggtccag accgccaccg cggcgccgtc gtccgcgacc cacaccttgc
    10021 cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc cgctcgatgt
    10081 ggcggtccgg atcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg gcggcgaggg
    10141 tgcgtacggc cctggggacg tcgtcgcggg tggcgaggcg caccgtgggc ttgtactcgg
    10201 tcatctcgag cctagggccg ggattctcct ccacgtcacc gcatgttaga agacttcctc
    10261 tgccctctcc gctgccagat ctctcgaggc cctgtgggag gaagagaaga ggtcagaagc
    10321 ttgccccact gtggggtgga ggggacagat aaaagtaccc agaaccagag ccacattaac
    10381 cggccctggg aatataaggt ggtcccagct cggggacaca ggatccctgg aggcagcaaa
    10441 catgctgtcc tgaagtggac ataggggccc gggttggagg aagaagacta gctgagctct
    10501 cggacccctg gaagatgcca tgacaggggg ctggaagagc tagcacagac tagagaggta
    10561 aggggggtag gggagctgcc caaatgaaag gagtgagagg tgacccgaat ccacaggaga
    10621 acggggtgtc caggcaaaga aagcaagagg atggagaggt ggctaaagcc agggagacgg
    10681 ggtactttgg ggttgtccag aaaaacggtg atgatgcagg cctacaagaa ggggaggcgg
    10741 gacgcaaggg agacatccgt cggagaaggc catcctaaga aacgagagat ggcacaggcc
    10801 ccagaaggag aaggaaaagg gaacccagcg agtgaagacg gcatggggtt gggtgaggga
    10861 ggagagatgc ccggagagga cccagacacg gggaggatcc gctcagagga catcacgtgg
    10921 tgcagcgccg agaaggaagt gctccggaaa gagcatcctt gggcagcaac acagcagaga
    10981 gcaaggggaa gagggagtgg aggaagacgg aacctgaagg aggcggcagg gaaggatctg
    11041 ggccagccgt agaggtgacc caggccacaa gctgcagaca gaaagcggca caggcccagg
    11101 ggagagaatg ctggtcagag aaagcaaagg gcgaattcgt ttaaacctgc aggactagtc
    11161 cctttagtga gggttaattc tgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
    11221 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
    11281 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
    11341 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
    11401 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
    11461 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
    11521 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
    11581 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
    11641 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
    11701 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
    11761 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
    11821 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
    11881 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
    11941 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
    12001 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
    12061 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
    12121 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
    12181 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
    12241 ctcacgttaa gggattttgg tcatgagaca ataaccctga taaatgcttc aataatattg
    12301 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
    12361 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
    12421 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
    12481 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
    12541 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
    12601 tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg atggcatgac
    12661 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
    12721 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
    12781 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
    12841 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
    12901 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
    12961 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
    13021 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
    13081 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
    13141 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
    13201 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
    13261 tgataatctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
    13321 tccgcgcaca tttccccgaa aagtgccacc tgatgcggtg tgaaataccg cacagatgcg
    13381 taaggagaaa ataccgcatc aggaaattgt aagcgttaat aattcagaag aactcgtcaa
    13441 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
    13501 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
    13561 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
    13621 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
    13681 cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
    13741 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
    13801 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
    13861 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
    13921 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
    13981 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca
    14041 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
    14101 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
    14161 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc
    14221 gaaacgatcc tcatcctgtc tcttgatcag agcttgatcc cctgcgccat cagatccttg
    14281 gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc
    14341 cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg
    14401 taagcccact gcaagctacc tgctttctct ttgcgcttgc gttttccctt gtccagatag
    14461 cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct ttctacgtga
    14521 aaaggatcta ggtgaagatc ctttttgata atctcatgcc tgacatttat attccccaga
    14581 acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg gctgagatca gccacttctt
    14641 ccccgataac ggagaccggc acactggcca tatcggtggt catcatgcgc cagctttcat
    14701 ccccgatatg caccaccggg taaagttcac gggagacttt atctgacagc agacgtgcac
    14761 tggccagggg gatcaccatc cgtcgccccg gcgtgtcaat aatatcactc tgtacatcca
    14821 caaacagacg ataacggctc tctcttttat aggtgtaaac cttaaactgc
//
