LOCUS       sequence-272068-       10981 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 10981)
  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 10981)
  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..10981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    77..913
                     /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    987..1008
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1023..1028,1029..1046,1047..1053)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1061..1077
                     /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     1085..1101
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1444..1463
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1462..1517
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1563..1582
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1692..2168
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     1950..1969
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    2200..2788
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2306..2317
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2715..2735
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2819..3565)
                     /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(3566..3589)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3589..3598
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3618..3829)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     promoter        3871..4238
                     /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    3879..3897
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3915..3933
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3951..3969
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3987..4005
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4023..4041
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4059..4077
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4095..4113
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             join(4245..4247,4248..4250,4251..4961)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4245..4247,4248..4250,4251..4961)
                     /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(4290..4311)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4551..4570)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4898..4919
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5151..5285
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     5229..5248
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    5310..5534
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5523..5540
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(5572..6171)
                     /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(5655..5675)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6152..6171
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(6181..6234)
                     /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    6258..6283
                     /label=SA
                     /note="splice acceptor site"
     misc_feature    6290..7093
                     /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(7132..7150)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7171..7187)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7195..7211
                     /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(7219..7225,7226..7243,7244..7249))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7264..7285
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7402..7419)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7573..8161)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7653..7672)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(8285..8353,8354..9145)
                     /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(8503..8522)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(9506..9525)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     10120..10139
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(10542..10844)
                     /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(10644..10663)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     join(10967..10981,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(10981,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg aatttagcgg
       61 ccgcgaattc gcccttagag cagagccagg aacccctgta gggaaggggc aggagagcca
      121 ggggcatgag atggtggacg aggaaggggg acagggaagc ctgagcgcct ctcctgggct
      181 tgccaaggac tcaaacccag aagcccagag cagggcctta gggaagcggg accctgctct
      241 gggcggagga atatgtccca gatagcactg gggactcttt aaggaaagaa ggatggagaa
      301 agagaaaggg agtagaggcg gccacgacct ggtgaacacc taggacgcac cattctcaca
      361 aagggagttt tccacacgga cacccccctc ctcaccacag ccctgccagg acggggctgg
      421 ctactggcct tatctcacag gtaaaactga cgcacggagg aacaatataa attggggact
      481 agaaaggtga agagccaaag ttagaactca ggaccaactt attctgattt tgtttttcca
      541 aactgcttct cctcttggga agtgtaagga agctgcagca ccaggatcag tgaaacgcac
      601 cagacagccg cgtcagagca gctcaggttc tgggagaggg tagcgcaggg tggccactga
      661 gaaccgggca ggtcacgcat cccccccttc cctcccaccc cctgccaagc tctccctccc
      721 aggatcctct ctggctccat cgtaagcaaa ccttagaggt tctggcaagg agagagatgg
      781 ctccaggaaa tgggggtgtg tcaccagata aggaatctgc ctaacaggag gtgggggtta
      841 gacccaatat caggagacta ggaaggagga ggcctaagga tggggctttt ctgtcaccaa
      901 tcctgtccct agtaaagctt agtactgtcg atccgctggc acgacaggtt tcccgactgg
      961 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     1021 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     1081 cacacaggaa acagctatga ccatgattac gccaagcttg ggctgcaggt cgagggatct
     1141 ccataagaga agagggacag ctatgactgg gagtagtcag gagaggagga aaaatctggc
     1201 tagtaaaaca tgtaaggaaa attttaggga tgttaaagaa aaaaataaca caaaacaaaa
     1261 tataaaaaaa atctaacctc aagtcaaggc ttttctatgg aataaggaat ggacagcagg
     1321 gggctgtttc atatactgat gacctcttta tagccaacct ttgttcatgg cagccagcat
     1381 atgggcatat gttgccaaac tctaaaccaa atactcattc tgatgtttta aatgatttgc
     1441 cctcccatat gtccttccga gtgagagaca caaaaaattc caacacacta ttgcaatgaa
     1501 aataaatttc ctttattagc cagaagtcag atgctcaagg ggcttcatga tgtccccata
     1561 atttttggca gagggaaaaa gatctcagtg gtatttgtga gccagggcat tggccacacc
     1621 agccaccacc ttctgatagg cagcctgcac ctgaggagtg aattcacgcg cctggttacc
     1681 tacaaaatca gaaggacagg gaagggagca gtggttcacg cctgtaatcc cagcaatttg
     1741 ggaggccaag gtgggtagat cacctgagat taggagttgg agaccagcct ggccaatatg
     1801 gtgaaacccc gtctctacca aaaaaacaaa aattagctga gcctggtcat gcatgcctgg
     1861 aatcccaaca actcgggagg ctgaggcagg agaatcgctt gaacccagga ggcggagatt
     1921 gcagtgagcc aagattgtgc cactgcactc cagcttggtt cccaatagac cccgcaggcc
     1981 ctacaggttg tcttcccaac ttgccccttg ctccatacca cccccctcca ccccataata
     2041 ttatagaagg acacctagtc agacaaaatg atgcaactta attttattag gacaaggctg
     2101 gtgggcactg gagtggcaac ttccagggcc aggagaggca ctggggaggg gtcacaggga
     2161 tgccacccgt agatctctcg agcagcgctc ggtatcgatg cggggaggcg gcccaaaggg
     2221 agatccgact cgtctgaggg cgaaggcgaa gacgcggaag aggccgcaga gccggcagca
     2281 ggccgcggga aggaaggtcc gctggattga gggccgaagg gacgtagcag aaggacgtcc
     2341 cgcgcagaat ccaggtggca acataggcga gcagccaagg aaaggacgat gatttccccg
     2401 acaacaccac ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc
     2461 aggcggcgat gagttccgcc gtggcaatag ggagggggaa agcgaaagtc ccggaaagga
     2521 gctgacaggt ggtggcaatg ccccaaccag tgggggttgc gtcagcaaac acagtgcaca
     2581 ccacgccacg ttgcctgaca acgggccaca actcctcata aagagacagc aaccaggatt
     2641 tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac aaaggcatta
     2701 aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca gtcaatcttt
     2761 cacaaatttt gtaatccaga ggttgattat cgataagctt gatatcgaat tcacgcgttt
     2821 acccggggag catgtcaagg tcaaaatcgt caagagcgtc agcaggcagc atatcaaggt
     2881 caaagtcgtc aagggcatcg gctgggagca tgtctaagtc aaaatcgtca agggcgtcgg
     2941 tcggcccgcc gctttcgcac tttagctgtt tctccaggcc acatatgatt agttccaggc
     3001 cgaaaaggaa ggcaggttcg gctccctgcc ggtcgaacag ctcaattgct tgtttcagaa
     3061 gtgggggcat agaatcggtg gtaggtgtct ctctttcctc ttttgctact tgatgctcct
     3121 gttcctccaa tacgcagccc agtgtaaagt ggcccacggc ggacagagcg tacagtgcgt
     3181 tctccaggga gaagccttgc tgacacagga acgcgagctg attttccagg gtttcgtact
     3241 gtttctctgt tgggcgggtg ccgagatgca ctttagcccc gtcgcgatgt gagaggagag
     3301 cacagcggta tgacttggcg ttgttccgca gaaagtcttg ccatgactcg ccttccaggg
     3361 ggcaggagtg ggtatgatgc ctgtccagca tctcgattgg cagggcatcg agcagggccc
     3421 gcttgttctt cacgtgccag tacagggtag gctgctcaac tcccagcttt tgagcgagtt
     3481 tccttgtcgt caggccttcg ataccgactc cattgagtaa ttccagagca gagtttatga
     3541 ctttgctctt gtccagtcta gacatcttat cgtcatcgtc tttgtaatcc atggtggcgg
     3601 atcccgcgtc acgacacctg tgttctggcg gcaaacccgt tgcgaaaaag aacgttcacg
     3661 gcgactactg cacttatata cggttctccc ccaccctcgg gaaaaaggcg gagccagtac
     3721 acgacatcac tttcccagtt taccccgcgc caccttctct aggcaccggt tcaattgccg
     3781 acccctcccc ccaacttctc ggggactgtg ggcgatgtgc gctctgccca ctgacgggca
     3841 ccggagccac tcgagtggaa ttatcacctc gagtttactc cctatcagtg atagagaacg
     3901 tatgaagagt ttactcccta tcagtgatag agaacgtatg cagactttac tccctatcag
     3961 tgatagagaa cgtataagga gtttactccc tatcagtgat agagaacgta tgaccagttt
     4021 actccctatc agtgatagag aacgtatcta cagtttactc cctatcagtg atagagaacg
     4081 tatatccagt ttactcccta tcagtgatag agaacgtata agctttgctt atgtaaacca
     4141 gggcgcctat aaaagagtgc tgattttttg agtaaacttc aattccacaa cacttttgtc
     4201 ttataccaac tttccgtacc acttcctacc ctcgtaaagg taccatggtg agcaagggcg
     4261 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     4321 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     4381 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     4441 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     4501 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     4561 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     4621 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     4681 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     4741 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     4801 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     4861 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     4921 ccgccgccgg gatcactctc ggcatggacg agctgtacaa ggaattctaa actagtagac
     4981 cacctcccct gcgagctaag ctggacagcc aatgacgggt aagagagtga catttttcac
     5041 taacctaaga caggagggcc gtcagagcta ctgcctaatc caaagacggg taaaagtgat
     5101 aaaaatgtat cactccaacc taagacaggc gcagcttccg agggatttga gatccagaca
     5161 tgataagata cattgatgag tttggacaaa ccaaaactag aatgcagtga aaaaaatgcc
     5221 ttatttgtga aatttgtgat gctattgcct tatttgtaac cattataagc tgcaataaac
     5281 aagttctagt caataatcaa tgtcgagacc catagagccc accgcatccc cagcatgcct
     5341 gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga
     5401 atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg
     5461 agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca
     5521 actagaaggc acagtcgagg ctgatcagcg ggtttaaacg ggccctctag atcaggcacc
     5581 gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc acctcgacgt cggcggtgac
     5641 ggtgaagccg agccgctcgt agaaggggag gttgcggggc gcggaggtct ccaggaaggc
     5701 gggcaccccg gcgcgctcgg ccgcctccac tccggggagc acgacggcgc tgcccagacc
     5761 cttgccctgg tggtcgggcg agacgccgac ggtggccagg aaccacgcgg gctccttggg
     5821 ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg gccagccggg aaccgctcaa
     5881 ctcggccatg cgcgggccga tctcggcgaa caccgccccc gcttcgacgc tctccggcgt
     5941 ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac accttgccga tgtcgagccc
     6001 gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc tcgatgtggc ggtccggatc
     6061 gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc gtacggccct
     6121 ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca tctcgagcct
     6181 agggccggga ttctcctcca cgtcaccgca tgttagaaga cttcctctgc cctctccgct
     6241 gccagatctc tcgaggccct gtgggaggaa gagaagaggt cagaagcttg ccccactgtg
     6301 gggtggaggg gacagataaa agtacccaga accagagcca cattaaccgg ccctgggaat
     6361 ataaggtggt cccagctcgg ggacacagga tccctggagg cagcaaacat gctgtcctga
     6421 agtggacata ggggcccggg ttggaggaag aagactagct gagctctcgg acccctggaa
     6481 gatgccatga cagggggctg gaagagctag cacagactag agaggtaagg ggggtagggg
     6541 agctgcccaa atgaaaggag tgagaggtga cccgaatcca caggagaacg gggtgtccag
     6601 gcaaagaaag caagaggatg gagaggtggc taaagccagg gagacggggt actttggggt
     6661 tgtccagaaa aacggtgatg atgcaggcct acaagaaggg gaggcgggac gcaagggaga
     6721 catccgtcgg agaaggccat cctaagaaac gagagatggc acaggcccca gaaggagaag
     6781 gaaaagggaa cccagcgagt gaagacggca tggggttggg tgagggagga gagatgcccg
     6841 gagaggaccc agacacgggg aggatccgct cagaggacat cacgtggtgc agcgccgaga
     6901 aggaagtgct ccggaaagag catccttggg cagcaacaca gcagagagca aggggaagag
     6961 ggagtggagg aagacggaac ctgaaggagg cggcagggaa ggatctgggc cagccgtaga
     7021 ggtgacccag gccacaagct gcagacagaa agcggcacag gcccagggga gagaatgctg
     7081 gtcagagaaa gcaaagggcg aattcgttta aacctgcagg actagctagt ccctttagtg
     7141 agggttaatt ctgagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     7201 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     7261 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     7321 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     7381 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     7441 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     7501 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7561 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7621 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7681 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7741 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7801 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7861 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7921 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7981 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     8041 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     8101 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     8161 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     8221 agggattttg gtcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     8281 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     8341 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     8401 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     8461 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     8521 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     8581 cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     8641 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     8701 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     8761 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     8821 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     8881 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     8941 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     9001 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     9061 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     9121 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     9181 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     9241 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     9301 atttccccga aaagtgccac ctgatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     9361 aataccgcat caggaaattg taagcgttaa taattcagaa gaactcgtca agaaggcgat
     9421 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
     9481 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
     9541 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
     9601 tgatattcgg caagcaggca tcgccatgca tgggtcacga cgagatcctc gccgtcgggc
     9661 atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc
     9721 agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt
     9781 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca
     9841 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc
     9901 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct
     9961 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca
    10021 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc
    10081 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc
    10141 ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac
    10201 gatcctcatc ctgtctcttg atcagagctt gatcccctgc gccatcagat ccttggcggc
    10261 aagaaagcca tccagtttac tttgcagggc ttcccaacct taccagaggg cgccccagct
    10321 ggcaattccg gttcgcttgc tgtccataaa accgcccagt ctagctatcg ccatgtaagc
    10381 ccactgcaag ctacctgctt tctctttgcg cttgcgtttt cccttgtcca gatagcccag
    10441 tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta cgtgaaaagg
    10501 atctaggtga agatcctttt tgataatctc atgcctgaca tttatattcc ccagaacatc
    10561 aggttaatgg cgtttttgat gtcattttcg cggtggctga gatcagccac ttcttccccg
    10621 ataacggaga ccggcacact ggccatatcg gtggtcatca tgcgccagct ttcatccccg
    10681 atatgcacca ccgggtaaag ttcacgggag actttatctg acagcagacg tgcactggcc
    10741 agggggatca ccatccgtcg ccccggcgtg tcaataatat cactctgtac atccacaaac
    10801 agacgataac ggctctctct tttataggtg taaaccttaa actgccgtac gtataggctg
    10861 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
    10921 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
    10981 t
//
