LOCUS       sequence-264414-       10989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  donor plasmid for inducible transgene expression in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10989)
  AUTHORS   Chang Y, Hellwarth PB, Randolph LN, Sun Y, Xing Y, Zhu W, Lian XL, 
            Bao X
  TITLE     Fluorescent indicators for continuous and lineage-specific reporting
            of cell-cycle phases in human pluripotent stem cells.
  JOURNAL   Biotechnol Bioeng. 2020 Apr 11. doi: 10.1002/bit.27352.
  PUBMED    32277708
REFERENCE   2  (bases 1 to 10989)
  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"; journalName: "Biotechnol 
            Bioeng. 2020 Apr 11. doi: 10.1002/bit.27352."
COMMENT     Sequence Label: AAVS1-Puro Xlone-eGFP
FEATURES             Location/Qualifiers
     source          1..10989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(3..21)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(42..58)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    66..82
                     /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(90..96,97..114,115..120))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    135..156
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(273..290)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(444..1032)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(524..543)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(1156..1224,1225..2016)
                     /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(1374..1393)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(2377..2396)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2991..3010
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(3413..3715)
                     /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(3515..3534)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     3838..3860
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3852..3869
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3853..3869
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3876..3895
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3876..3894
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3935..4771
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     protein_bind    4845..4866
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4881..4886,4887..4904,4905..4911)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4919..4935
                     /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)."
     primer_bind     4943..4959
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5302..5321
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    5320..5375
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     5421..5440
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5550..6026
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     5808..5827
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    6058..6646
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6164..6175
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6573..6593
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6677..7423)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             complement(7424..7447)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      7447..7456
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7476..7687)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     promoter        7729..8096
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    7737..7755
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7773..7791
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7809..7827
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7845..7863
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7881..7899
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7917..7935
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7953..7971
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      8103..8112
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(8109..8111,8112..8114,8115..8825)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(8109..8111,8112..8114,8115..8825)
                     /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(8154..8175)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8415..8434)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8762..8783
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    9015..9149
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     9093..9112
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    9174..9398
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     9387..9404
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(9436..10035)
                     /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(9519..9539)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     10016..10035
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(10045..10098)
                     /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    10122..10147
                     /label=SA
                     /note="splice acceptor site"
     misc_feature    10154..10957
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
ORIGIN
        1 accctttagt gagggttaat tctgagcttg gcgtaatcat ggtcatagct gtttcctgtg
       61 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
      121 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
      181 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      241 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
      301 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
      361 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      421 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      481 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      541 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      601 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
      661 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
      721 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
      781 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
      841 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
      901 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
      961 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1021 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1081 aactcacgtt aagggatttt ggtcatgaga caataaccct gataaatgct tcaataatat
     1141 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     1201 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1261 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     1321 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     1381 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     1441 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     1501 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     1561 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     1621 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     1681 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     1741 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     1801 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     1861 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     1921 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     1981 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     2041 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     2101 tttgataatc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     2161 gttccgcgca catttccccg aaaagtgcca cctgatgcgg tgtgaaatac cgcacagatg
     2221 cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta ataattcaga agaactcgtc
     2281 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
     2341 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
     2401 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
     2461 attttccacc atgatattcg gcaagcaggc atcgccatgc atgggtcacg acgagatcct
     2521 cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat
     2581 gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct
     2641 cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc
     2701 gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga
     2761 gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt
     2821 cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt
     2881 cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct
     2941 gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat
     3001 agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa
     3061 tcatgcgaaa cgatcctcat cctgtctctt gatcagagct tgatcccctg cgccatcaga
     3121 tccttggcgg caagaaagcc atccagttta ctttgcaggg cttcccaacc ttaccagagg
     3181 gcgccccagc tggcaattcc ggttcgcttg ctgtccataa aaccgcccag tctagctatc
     3241 gccatgtaag cccactgcaa gctacctgct ttctctttgc gcttgcgttt tcccttgtcc
     3301 agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct
     3361 acgtgaaaag gatctaggtg aagatccttt ttgataatct catgcctgac atttatattc
     3421 cccagaacat caggttaatg gcgtttttga tgtcattttc gcggtggctg agatcagcca
     3481 cttcttcccc gataacggag accggcacac tggccatatc ggtggtcatc atgcgccagc
     3541 tttcatcccc gatatgcacc accgggtaaa gttcacggga gactttatct gacagcagac
     3601 gtgcactggc cagggggatc accatccgtc gccccggcgt gtcaataata tcactctgta
     3661 catccacaaa cagacgataa cggctctctc ttttataggt gtaaacctta aactgccgta
     3721 cgtataggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     3781 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     3841 agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat
     3901 tgaatttagc ggccgcgaat tcgcccttgc ccttagagca gagccaggaa cccctgtagg
     3961 gaaggggcag gagagccagg ggcatgagat ggtggacgag gaagggggac agggaagcct
     4021 gagcgcctct cctgggcttg ccaaggactc aaacccagaa gcccagagca gggccttagg
     4081 gaagcgggac cctgctctgg gcggaggaat atgtcccaga tagcactggg gactctttaa
     4141 ggaaagaagg atggagaaag agaaagggag tagaggcggc cacgacctgg tgaacaccta
     4201 ggacgcacca ttctcacaaa gggagttttc cacacggaca cccccctcct caccacagcc
     4261 ctgccaggac ggggctggct actggcctta tctcacaggt aaaactgacg cacggaggaa
     4321 caatataaat tggggactag aaaggtgaag agccaaagtt agaactcagg accaacttat
     4381 tctgattttg tttttccaaa ctgcttctcc tcttgggaag tgtaaggaag ctgcagcacc
     4441 aggatcagtg aaacgcacca gacagccgcg tcagagcagc tcaggttctg ggagagggta
     4501 gcgcagggtg gccactgaga accgggcagg tcacgcatcc cccccttccc tcccaccccc
     4561 tgccaagctc tccctcccag gatcctctct ggctccatcg taagcaaacc ttagaggttc
     4621 tggcaaggag agagatggct ccaggaaatg ggggtgtgtc accagataag gaatctgcct
     4681 aacaggaggt gggggttaga cccaatatca ggagactagg aaggaggagg cctaaggatg
     4741 gggcttttct gtcaccaatc ctgtccctag taaagcttag tactgtcgat ccgctggcac
     4801 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     4861 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     4921 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttggg
     4981 ctgcaggtcg agggatctcc ataagagaag agggacagct atgactggga gtagtcagga
     5041 gaggaggaaa aatctggcta gtaaaacatg taaggaaaat tttagggatg ttaaagaaaa
     5101 aaataacaca aaacaaaata taaaaaaaat ctaacctcaa gtcaaggctt ttctatggaa
     5161 taaggaatgg acagcagggg gctgtttcat atactgatga cctctttata gccaaccttt
     5221 gttcatggca gccagcatat gggcatatgt tgccaaactc taaaccaaat actcattctg
     5281 atgttttaaa tgatttgccc tcccatatgt ccttccgagt gagagacaca aaaaattcca
     5341 acacactatt gcaatgaaaa taaatttcct ttattagcca gaagtcagat gctcaagggg
     5401 cttcatgatg tccccataat ttttggcaga gggaaaaaga tctcagtggt atttgtgagc
     5461 cagggcattg gccacaccag ccaccacctt ctgataggca gcctgcacct gaggagtgaa
     5521 ttcacgcgcc tggttaccta caaaatcaga aggacaggga agggagcagt ggttcacgcc
     5581 tgtaatccca gcaatttggg aggccaaggt gggtagatca cctgagatta ggagttggag
     5641 accagcctgg ccaatatggt gaaaccccgt ctctaccaaa aaaacaaaaa ttagctgagc
     5701 ctggtcatgc atgcctggaa tcccaacaac tcgggaggct gaggcaggag aatcgcttga
     5761 acccaggagg cggagattgc agtgagccaa gattgtgcca ctgcactcca gcttggttcc
     5821 caatagaccc cgcaggccct acaggttgtc ttcccaactt gccccttgct ccataccacc
     5881 cccctccacc ccataatatt atagaaggac acctagtcag acaaaatgat gcaacttaat
     5941 tttattagga caaggctggt gggcactgga gtggcaactt ccagggccag gagaggcact
     6001 ggggaggggt cacagggatg ccacccgtag atctctcgag cagcgctcgg tatcgatgcg
     6061 gggaggcggc ccaaagggag atccgactcg tctgagggcg aaggcgaaga cgcggaagag
     6121 gccgcagagc cggcagcagg ccgcgggaag gaaggtccgc tggattgagg gccgaaggga
     6181 cgtagcagaa ggacgtcccg cgcagaatcc aggtggcaac ataggcgagc agccaaggaa
     6241 aggacgatga tttccccgac aacaccacgg aattgtcagt gcccaacagc cgagcccctg
     6301 tccagcagcg ggcaaggcag gcggcgatga gttccgccgt ggcaataggg agggggaaag
     6361 cgaaagtccc ggaaaggagc tgacaggtgg tggcaatgcc ccaaccagtg ggggttgcgt
     6421 cagcaaacac agtgcacacc acgccacgtt gcctgacaac gggccacaac tcctcataaa
     6481 gagacagcaa ccaggattta tacaaggagg agaaaatgaa agccatacgg gaagcaatag
     6541 catgatacaa aggcattaaa gcagcgtatc cacatagcgt aaaaggagca acatagttaa
     6601 gaataccagt caatctttca caaattttgt aatccagagg ttgattatcg ataagcttga
     6661 tatcgaattc acgcgtttac ccggggagca tgtcaaggtc aaaatcgtca agagcgtcag
     6721 caggcagcat atcaaggtca aagtcgtcaa gggcatcggc tgggagcatg tctaagtcaa
     6781 aatcgtcaag ggcgtcggtc ggcccgccgc tttcgcactt tagctgtttc tccaggccac
     6841 atatgattag ttccaggccg aaaaggaagg caggttcggc tccctgccgg tcgaacagct
     6901 caattgcttg tttcagaagt gggggcatag aatcggtggt aggtgtctct ctttcctctt
     6961 ttgctacttg atgctcctgt tcctccaata cgcagcccag tgtaaagtgg cccacggcgg
     7021 acagagcgta cagtgcgttc tccagggaga agccttgctg acacaggaac gcgagctgat
     7081 tttccagggt ttcgtactgt ttctctgttg ggcgggtgcc gagatgcact ttagccccgt
     7141 cgcgatgtga gaggagagca cagcggtatg acttggcgtt gttccgcaga aagtcttgcc
     7201 atgactcgcc ttccaggggg caggagtggg tatgatgcct gtccagcatc tcgattggca
     7261 gggcatcgag cagggcccgc ttgttcttca cgtgccagta cagggtaggc tgctcaactc
     7321 ccagcttttg agcgagtttc cttgtcgtca ggccttcgat accgactcca ttgagtaatt
     7381 ccagagcaga gtttatgact ttgctcttgt ccagtctaga catcttatcg tcatcgtctt
     7441 tgtaatccat ggtggcggat cccgcgtcac gacacctgtg ttctggcggc aaacccgttg
     7501 cgaaaaagaa cgttcacggc gactactgca cttatatacg gttctccccc accctcggga
     7561 aaaaggcgga gccagtacac gacatcactt tcccagttta ccccgcgcca ccttctctag
     7621 gcaccggttc aattgccgac ccctcccccc aacttctcgg ggactgtggg cgatgtgcgc
     7681 tctgcccact gacgggcacc ggagccactc gagtggaatt atcacctcga gtttactccc
     7741 tatcagtgat agagaacgta tgaagagttt actccctatc agtgatagag aacgtatgca
     7801 gactttactc cctatcagtg atagagaacg tataaggagt ttactcccta tcagtgatag
     7861 agaacgtatg accagtttac tccctatcag tgatagagaa cgtatctaca gtttactccc
     7921 tatcagtgat agagaacgta tatccagttt actccctatc agtgatagag aacgtataag
     7981 ctttgcttat gtaaaccagg gcgcctataa aagagtgctg attttttgag taaacttcaa
     8041 ttccacaaca cttttgtctt ataccaactt tccgtaccac ttcctaccct cgtaaaggta
     8101 ccgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     8161 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     8221 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     8281 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     8341 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     8401 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     8461 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     8521 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     8581 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     8641 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     8701 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     8761 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     8821 acaaggaatt ctaaactagt agaccacctc ccctgcgagc taagctggac agccaatgac
     8881 gggtaagaga gtgacatttt tcactaacct aagacaggag ggccgtcaga gctactgcct
     8941 aatccaaaga cgggtaaaag tgataaaaat gtatcactcc aacctaagac aggcgcagct
     9001 tccgagggat ttgagatcca gacatgataa gatacattga tgagtttgga caaaccaaaa
     9061 ctagaatgca gtgaaaaaaa tgccttattt gtgaaatttg tgatgctatt gccttatttg
     9121 taaccattat aagctgcaat aaacaagttc tagtcaataa tcaatgtcga gacccataga
     9181 gcccaccgca tccccagcat gcctgctatt gtcttcccaa tcctccccct tgctgtcctg
     9241 ccccacccca ccccccagaa tagaatgaca cctactcaga caatgcgatg caatttcctc
     9301 attttattag gaaaggacag tgggagtggc accttccagg gtcaaggaag gcacggggga
     9361 ggggcaaaca acagatggct ggcaactaga aggcacagtc gaggctgatc agcgggttta
     9421 aacgggccct ctagatcagg caccgggctt gcgggtcatg caccaggtgc gcggtccttc
     9481 gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc tcgtagaagg ggaggttgcg
     9541 gggcgcggag gtctccagga aggcgggcac cccggcgcgc tcggccgcct ccactccggg
     9601 gagcacgacg gcgctgccca gacccttgcc ctggtggtcg ggcgagacgc cgacggtggc
     9661 caggaaccac gcgggctcct tgggccggtg cggcgccagg aggccttcca tctgttgctg
     9721 cgcggccagc cgggaaccgc tcaactcggc catgcgcggg ccgatctcgg cgaacaccgc
     9781 ccccgcttcg acgctctccg gcgtggtcca gaccgccacc gcggcgccgt cgtccgcgac
     9841 ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag agttcttgca gctcggtgac
     9901 ccgctcgatg tggcggtccg gatcgacggt gtggcgcgtg gcggggtagt cggcgaacgc
     9961 ggcggcgagg gtgcgtacgg ccctggggac gtcgtcgcgg gtggcgaggc gcaccgtggg
    10021 cttgtactcg gtcatctcga gcctagggcc gggattctcc tccacgtcac cgcatgttag
    10081 aagacttcct ctgccctctc cgctgccaga tctctcgagg ccctgtggga ggaagagaag
    10141 aggtcagaag cttgccccac tgtggggtgg aggggacaga taaaagtacc cagaaccaga
    10201 gccacattaa ccggccctgg gaatataagg tggtcccagc tcggggacac aggatccctg
    10261 gaggcagcaa acatgctgtc ctgaagtgga cataggggcc cgggttggag gaagaagact
    10321 agctgagctc tcggacccct ggaagatgcc atgacagggg gctggaagag ctagcacaga
    10381 ctagagaggt aaggggggta ggggagctgc ccaaatgaaa ggagtgagag gtgacccgaa
    10441 tccacaggag aacggggtgt ccaggcaaag aaagcaagag gatggagagg tggctaaagc
    10501 cagggagacg gggtactttg gggttgtcca gaaaaacggt gatgatgcag gcctacaaga
    10561 aggggaggcg ggacgcaagg gagacatccg tcggagaagg ccatcctaag aaacgagaga
    10621 tggcacaggc cccagaagga gaaggaaaag ggaacccagc gagtgaagac ggcatggggt
    10681 tgggtgaggg aggagagatg cccggagagg acccagacac ggggaggatc cgctcagagg
    10741 acatcacgtg gtgcagcgcc gagaaggaag tgctccggaa agagcatcct tgggcagcaa
    10801 cacagcagag agcaagggga agagggagtg gaggaagacg gaacctgaag gaggcggcag
    10861 ggaaggatct gggccagccg tagaggtgac ccaggccaca agctgcagac agaaagcggc
    10921 acaggcccag gggagagaat gctggtcaga gaaagcaaag ggcgaattcg tttaaacctg
    10981 caggactag
//
