LOCUS       sequence-313586-       10814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAVS1 donor plasmid for targeted inducible eGFP expression in human 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10814)
  AUTHORS   Chang Y, Syahirah R, Oprescu SN, Wang X, Jung J, Cooper SH, 
            Torregrosa-Allen S, Elzey BD, Hsu AY, Randolph LN, Sun Y, Kuang S, 
            Broxmeyer HE, Deng Q, Lian X, Bao X
  TITLE     Chemically-defined generation of human hemogenic endothelium and 
            definitive hematopoietic progenitor cells.
  JOURNAL   Biomaterials. 2022 May 6;285:121569. doi: 
            10.1016/j.biomaterials.2022.121569.
  PUBMED    35567999
REFERENCE   2  (bases 1 to 10814)
  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: 
            "Biomaterials."; date: "2022-05-6"; volume: "285:121569. doi"; 
            pages: " 10.1016/j.biomaterials.2022.121569"
COMMENT     Sequence Label: AAVS1-Blasticidin XLone-eGFP
FEATURES             Location/Qualifiers
     source          1..10814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     135..154
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     394..415
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    592..610
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    628..646
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    664..682
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    700..718
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    736..754
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    772..790
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        840..1051
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      1071..1080
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1080..1103
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1104..1850
                     /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"
     misc_feature    1881..2469
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1934..1954)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2352..2363)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2501..2977
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2700..2719)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3087..3106)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3152..3207
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3206..3225)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3568..3584)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3592..3608
                     /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(3616..3622,3623..3640,3641..3646))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3661..3682
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    3756..4592
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(4626..4645)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4627..4645)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4652..4669)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4652..4668)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4661..4683)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             4806..5108
                     /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     4987..5006
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(5511..5530)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6125..6144
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(6505..7296,7297..7365))
                     /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     7128..7147
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7489..8077
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7978..7997
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8231..8248
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8365..8386
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8401..8406,8407..8424,8425..8431)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8439..8455
                     /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     8463..8479
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8500..8518
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    8557..9360
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     misc_feature    9367..9392
                     /label=SA
                     /note="splice acceptor site"
     CDS             9416..9469
                     /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"
     primer_bind     complement(9942..9959)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9948..10172
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    10234..10368
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10271..10290)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(10558..10814,1..454,455..457,458..460))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(10558..10814,1..454,455..457,458..460))
                     /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(10600..10621)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
       61 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
      121 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
      181 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
      241 ggctgaagca ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca
      301 gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
      361 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
      421 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtaccttta cgagggtagg
      481 aagtggtacg gaaagttggt ataagacaaa agtgttgtgg aattgaagtt tactcaaaaa
      541 atcagcactc ttttataggc gccctggttt acataagcaa agcttatacg ttctctatca
      601 ctgataggga gtaaactgga tatacgttct ctatcactga tagggagtaa actgtagata
      661 cgttctctat cactgatagg gagtaaactg gtcatacgtt ctctatcact gatagggagt
      721 aaactcctta tacgttctct atcactgata gggagtaaac tcttcatacg ttctctatca
      781 ctgataggga gtaaactcga ggtgataatt ccactcgagt ggctccggtg cccgtcagtg
      841 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
      901 cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
      961 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
     1021 ttttcgcaac gggtttgccg ccagaacaca ggtgtcgtga cgcgggatcc gccaccatgg
     1081 attacaaaga cgatgacgat aagatgtcta gactggacaa gagcaaagtc ataaactctg
     1141 ctctggaatt actcaatgga gtcggtatcg aaggcctgac gacaaggaaa ctcgctcaaa
     1201 agctgggagt tgagcagcct accctgtact ggcacgtgaa gaacaagcgg gccctgctcg
     1261 atgccctgcc aatcgagatg ctggacaggc atcataccca ctcctgcccc ctggaaggcg
     1321 agtcatggca agactttctg cggaacaacg ccaagtcata ccgctgtgct ctcctctcac
     1381 atcgcgacgg ggctaaagtg catctcggca cccgcccaac agagaaacag tacgaaaccc
     1441 tggaaaatca gctcgcgttc ctgtgtcagc aaggcttctc cctggagaac gcactgtacg
     1501 ctctgtccgc cgtgggccac tttacactgg gctgcgtatt ggaggaacag gagcatcaag
     1561 tagcaaaaga ggaaagagag acacctacca ccgattctat gcccccactt ctgaaacaag
     1621 caattgagct gttcgaccgg cagggagccg aacctgcctt ccttttcggc ctggaactaa
     1681 tcatatgtgg cctggagaaa cagctaaagt gcgaaagcgg cgggccgacc gacgcccttg
     1741 acgattttga cttagacatg ctcccagccg atgcccttga cgactttgac cttgatatgc
     1801 tgcctgctga cgctcttgac gattttgacc ttgacatgct ccccgggtaa acgcgtgaat
     1861 tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     1921 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     1981 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     2041 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     2101 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     2161 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     2221 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     2281 cgtcctttcc ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct
     2341 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     2401 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     2461 cctccccgca tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc
     2521 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct
     2581 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg
     2641 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt
     2701 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc
     2761 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat
     2821 gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag
     2881 gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg
     2941 attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccaggcg
     3001 cgtgaattca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
     3061 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
     3121 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
     3181 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
     3241 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
     3301 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
     3361 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
     3421 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
     3481 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac
     3541 ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     3601 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3661 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3721 cctgtcgtgc cagcggatcg acagtactaa gctttactag ggacaggatt ggtgacagaa
     3781 aagccccatc cttaggcctc ctccttccta gtctcctgat attgggtcta acccccacct
     3841 cctgttaggc agattcctta tctggtgaca cacccccatt tcctggagcc atctctctcc
     3901 ttgccagaac ctctaaggtt tgcttacgat ggagccagag aggatcctgg gagggagagc
     3961 ttggcagggg gtgggaggga agggggggat gcgtgacctg cccggttctc agtggccacc
     4021 ctgcgctacc ctctcccaga acctgagctg ctctgacgcg gctgtctggt gcgtttcact
     4081 gatcctggtg ctgcagcttc cttacacttc ccaagaggag aagcagtttg gaaaaacaaa
     4141 atcagaataa gttggtcctg agttctaact ttggctcttc acctttctag tccccaattt
     4201 atattgttcc tccgtgcgtc agttttacct gtgagataag gccagtagcc agccccgtcc
     4261 tggcagggct gtggtgagga ggggggtgtc cgtgtggaaa actccctttg tgagaatggt
     4321 gcgtcctagg tgttcaccag gtcgtggccg cctctactcc ctttctcttt ctccatcctt
     4381 ctttccttaa agagtcccca gtgctatctg ggacatattc ctccgcccag agcagggtcc
     4441 cgcttcccta aggccctgct ctgggcttct gggtttgagt ccttggcaag cccaggagag
     4501 gcgctcaggc ttccctgtcc cccttcctcg tccaccatct catgcccctg gctctcctgc
     4561 cccttcccta caggggttcc tggctctgct ctaagggcga attcgcggcc gctaaattca
     4621 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
     4681 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     4741 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctatacg
     4801 tacggcagtt taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg
     4861 tacagagtga tattattgac acgccggggc gacggatggt gatccccctg gccagtgcac
     4921 gtctgctgtc agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa
     4981 gctggcgcat gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag
     5041 tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg
     5101 gaatataaat gtcaggcatg agattatcaa aaaggatctt cacctagatc cttttcacgt
     5161 agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct
     5221 ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc
     5281 gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc
     5341 cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga
     5401 tctgatggcg caggggatca agctctgatc aagagacagg atgaggatcg tttcgcatga
     5461 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5521 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5581 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     5641 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5701 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5761 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5821 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5881 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5941 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     6001 aggatctcgt cgtgacccat gcatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6061 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6121 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     6181 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6241 gacgagttct tctgaattat taacgcttac aatttcctga tgcggtattt tctccttacg
     6301 catctgtgcg gtatttcaca ccgcatcagg tggcactttt cggggaaatg tgcgcggaac
     6361 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gattatcaaa
     6421 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6481 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6541 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6601 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6661 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6721 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6781 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6841 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6901 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6961 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7021 catgccatcc gtaagatgct tttctgtgac tggtgaatac tcaaccaagt cattctgaga
     7081 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7141 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7201 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7261 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7321 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7381 atattattga agcatttatc agggttattg tctcatgacc aaaatccctt aacgtgagtt
     7441 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7501 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7561 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7621 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7681 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7741 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7801 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7861 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7921 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7981 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     8041 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     8101 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     8161 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     8221 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     8281 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     8341 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     8401 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     8461 cacaggaaac agctatgacc atgattacgc caagctcaga attaaccctc actaaaggga
     8521 ctagctagtc ctgcaggttt aaacgaattc gccctttgct ttctctgacc agcattctct
     8581 cccctgggcc tgtgccgctt tctgtctgca gcttgtggcc tgggtcacct ctacggctgg
     8641 cccagatcct tccctgccgc ctccttcagg ttccgtcttc ctccactccc tcttcccctt
     8701 gctctctgct gtgttgctgc ccaaggatgc tctttccgga gcacttcctt ctcggcgctg
     8761 caccacgtga tgtcctctga gcggatcctc cccgtgtctg ggtcctctcc gggcatctct
     8821 cctccctcac ccaaccccat gccgtcttca ctcgctgggt tcccttttcc ttctccttct
     8881 ggggcctgtg ccatctctcg tttcttagga tggccttctc cgacggatgt ctcccttgcg
     8941 tcccgcctcc ccttcttgta ggcctgcatc atcaccgttt ttctggacaa ccccaaagta
     9001 ccccgtctcc ctggctttag ccacctctcc atcctcttgc tttctttgcc tggacacccc
     9061 gttctcctgt ggattcgggt cacctctcac tcctttcatt tgggcagctc ccctaccccc
     9121 cttacctctc tagtctgtgc tagctcttcc agccccctgt catggcatct tccaggggtc
     9181 cgagagctca gctagtcttc ttcctccaac ccgggcccct atgtccactt caggacagca
     9241 tgtttgctgc ctccagggat cctgtgtccc cgagctggga ccaccttata ttcccagggc
     9301 cggttaatgt ggctctggtt ctgggtactt ttatctgtcc cctccacccc acagtggggc
     9361 aagcttctga cctcttctct tcctcccaca gggcctcgag agatctggca gcggagaggg
     9421 cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggcccta ggaccggtat
     9481 gaaaacattt aacatttctc aacaagatct agaattagta gaagtagcga cagagaagat
     9541 tacaatgctt tatgaggata ataaacatca tgtgggagcg gcaattcgta cgaaaacagg
     9601 agaaatcatt tcggcagtac atattgaagc gtatatagga cgagtaactg tttgtgcaga
     9661 agccattgcg attggtagtg cagtttcgaa tggacaaaag gattttgaca cgattgtagc
     9721 tgttagacac ccttattctg acgaagtaga tagaagtatt cgagtggtaa gtccttgtgg
     9781 tatgtgtagg gagttgattt cagactatgc accagattgt tttgtgttaa tagaaatgaa
     9841 tggcaagtta gtcaaaacta cgattgaaga actcattcca ctcaaatata cccgaaatta
     9901 atgagaattc ggggatcaat tctctagagc tcgctgatca gcctcgactg tgccttctag
     9961 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
    10021 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
    10081 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
    10141 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
    10201 ctcgactaga gcttgcggaa cccttaatgg tacaacttgt ttattgcagc ttataatggt
    10261 tacaaataag gcaatagcat cacaaatttc acaaataagg catttttttc actgcattct
    10321 agttttggtt tgtccaaact catcaatgta tcttatcatg tctggatctc aaatccctcg
    10381 gaagctgcgc ctgtcttagg ttggagtgat acatttttat cacttttacc cgtctttgga
    10441 ttaggcagta gctctgacgg ccctcctgtc ttaggttagt gaaaaatgtc actctcttac
    10501 ccgtcattgg ctgtccagct tagctcgcag gggaggtggt ctactagttt agaattcctt
    10561 gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat
    10621 gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac tgggtgctca ggtagtggtt
    10681 gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag
    10741 ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga tgccgttctt
    10801 ctgcttgtcg gcca
//
