LOCUS       sequence-370987-       12058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A CASH-1 GSH targeting, doxycycling inducible humanized Oct4, Sox2, 
            and Klf4 expressing CRISPR/Cas9 donor plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12058)
  AUTHORS   Javaid N, Choi S
  TITLE     An Alternate Approach to Generate Induced Pluripotent Stem Cells 
            with Precise CRISPR/Cas9 Tool.
  JOURNAL   Stem Cells Int. 2022 Sep 22;2022:4537335. doi: 10.1155/2022/4537335.
            eCollection 2022.
  PUBMED    36187228
REFERENCE   2  (bases 1 to 12058)
  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: "Stem Cells 
            Int."; date: "2022-09-22"; volume: "2022:4537335. doi"; pages: " 
            10.1155/2022/4537335. eCollection 2022"
COMMENT     Sequence Label: pTetO-hOSK-CASH-1
FEATURES             Location/Qualifiers
     source          1..12058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1203..1236)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1247..1381
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1391..2137)
                     /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"
     promoter        complement(2157..2656)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2205..2224)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     2598..2621
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     complement(2683..2702)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2727..2775
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        2949..3316
                     /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    2957..2975
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2993..3011
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3029..3047
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3065..3083
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3101..3119
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3137..3155
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3173..3191
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             5432..5485
                     /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             6962..7021
                     /codon_start=1
                     /product="2A peptide from equine rhinitis A virus 
                     polyprotein"
                     /label=E2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="QCTNYALLKLAGDVESNPGP"
     CDS             join(7040..7042,7043..7045,7046..7759)
                     /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(7040..7042,7043..7045,7046..7756)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7085..7106)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7346..7365)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7693..7714
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    7775..7909
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    7912..7945
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     polyA_signal    7946..8170
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(8171..8569)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      8566..8575
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(8584..8913)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8598..8733
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8652..8671)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     8898..8918
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     protein_bind    8923..8956
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    complement(8957..8990)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(9837..9853)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9850..9872)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9861..9877
                     /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(9885..9891,9892..9909,9910..9915))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9930..9951
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10068..10085)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10239..10827)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10319..10338)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10998..11789,11790..11858))
                     /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     11621..11640
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11859..11963)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     12031..12049
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accagatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtaaata aatagtttaa tttaaaattc
      421 acatactgca gtataacata aggttcttgt ttgaatttta ctgtttgcca tgcattaaaa
      481 agcactcatt ttaaggctat ggaagatgct cacgtagtat ctctaagcac ctatttttca
      541 gaaggatttg tggcttctgt gccacagatg ataaggaatt ttattgttct ggcttccata
      601 ctcagcttga cctattccat cctaaataag ctaatcctgg agagagtttt gcttcaagtg
      661 taaaatgatc aacacaagaa tggaaagtaa atctaccacc acacatgtaa atactgatat
      721 ataggccata gtagagcctt atattcatat aaataaagtt tcagatcata cttctcccta
      781 aaatcaagac atccaatttt aagaacttct gtcctcatca ttggtaacga gtaccagaaa
      841 ctgatgttac agcatataaa atagttcttt ttaaatatca tgcaactctt accaatgttt
      901 tgattcagtg ataaaattat ggagcttcat aataaccagg aaaatgcagt gttaaaaaca
      961 acaataaaag gctagcaaga aattataaat atgttgttaa gggcattggc ttctttaggg
     1021 cttgctctcg taactgatac caaatgggaa ttcagagtct aaaaatatct tctttggtaa
     1081 aaggaaagtc ttggccagga agtaaataag cttttccctt aattcagcac atctccctaa
     1141 atacactatt atcacctacc atctggaatg aagcttccac caactaatga gctgccctca
     1201 gcataacttc gtatagcata cattatacga agttatgcga tcgcgagatc cagacatgat
     1261 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1321 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1381 tgggccatgg ttacccgggg agcatgtcaa ggtcaaaatc gtcaagagcg tcagcaggca
     1441 gcatatcaag gtcaaagtcg tcaagggcat cggctgggag catgtctaag tcaaaatcgt
     1501 caagggcgtc ggtcggcccg ccgctttcgc actttagctg tttctccagg ccacatatga
     1561 ttagttccag gccgaaaagg aaggcaggtt cggctccctg ccggtcgaac agctcaattg
     1621 cttgtttcag aagtgggggc atagaatcgg tggtaggtgt ctctctttcc tcttttgcta
     1681 cttgatgctc ctgttcctcc aatacgcagc ccagtgtaaa gtggcccacg gcggacagag
     1741 cgtacagtgc gttctccagg gagaagcctt gctgacacag gaacgcgagc tgattttcca
     1801 gggtttcgta ctgtttctct gttgggcggg tgccgagatg cactttagcc ccgtcgcgat
     1861 gtgagaggag agcacagcgg tatgacttgg cgttgttccg cagaaagtct tgccatgact
     1921 cgccttccag ggggcaggag tgggtatgat gcctgtccag catctcgatt ggcagggcat
     1981 cgagcagggc ccgcttgttc ttcacgtgcc agtacagggt aggctgctca actcccagct
     2041 tttgagcgag tttccttgtc gtcaggcctt cgataccgac tccattgagt aattccagag
     2101 cagagtttat gactttgctc ttgtccagtc tagacatggt ggcggcagat ctaggtcgaa
     2161 aggcccggag atgaggaaga ggagaacagc gcggcagacg tgcgcttttg aagcgtgcag
     2221 aatgccgggc ctccggagga ccttcgggcg cccgccccgc ccctgagccc gcccctgagc
     2281 ccgcccccgg acccacccct tcccagcctc tgagcccaga aagcgaagga gcaaagctgc
     2341 tattggccgc tgccccaaag gcctacccgc ttccattgct cagcggtgct gtccatctgc
     2401 acgagactag tgagacgtgc tacttccatt tgtcacgtcc tgcacgacgc gagctgcggg
     2461 gcggggggga acttcctgac taggggagga gtagaaggtg gcgcgaaggg gccaccaaag
     2521 aacggagccg gttggcgcct accggtggat gtggaatgtg tgcgaggcca gaggccactt
     2581 gtgtagcgcc aagtgcccag cggggctgct aaagcgcatg ctccagactg ccttgggaaa
     2641 agcgcctccc ctacccggta gaacgtacgg ggggaggcta actgaaacac ggaaggagac
     2701 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg
     2761 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     2821 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     2881 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     2941 cgccggcgga gtttactccc tatcagtgat agagaacgta tgaagagttt actccctatc
     3001 agtgatagag aacgtatgca gactttactc cctatcagtg atagagaacg tataaggagt
     3061 ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa
     3121 cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc
     3181 agtgatagag aacgtataag ctttgcttat gtaaaccagg gcgcctataa aagagtgctg
     3241 attttttgag taaacttcaa ttccacaaca cttttgtctt ataccaactt tccgtaccac
     3301 ttcctaccct cgtaaagccg ccaccatcga tatggcggga cacctggctt cagattttgc
     3361 cttctcgccc cctccaggtg gtggaggtga tgggccaggg gggccggagc cgggctgggt
     3421 tgatcctcgg acctggctaa gcttccaagg ccctcctgga gggccaggaa tcgggccggg
     3481 ggttgggcca ggctctgagg tgtgggggat tcccccatgc cccccgccgt atgagttctg
     3541 tggggggatg gcgtactgtg ggccccaggt tggagtgggg ctagtgcccc aaggcggctt
     3601 ggagacctct cagcctgagg gcgaagcagg agtcggggtg gagagcaact ccgatggggc
     3661 ctccccggag ccctgcaccg tcacccctgg tgccgtgaag ctggagaagg agaagctgga
     3721 gcaaaacccg gaggagtccc aggacatcaa agctctgcag aaagaactcg agcaatttgc
     3781 caagctcctg aagcagaaga ggatcaccct gggatataca caggccgatg tggggctcac
     3841 cctgggggtt ctatttggga aggtattcag ccaaacgacc atctgccgct ttgaggctct
     3901 gcagcttagc ttcaagaaca tgtgtaagct gcggcccttg ctgcagaagt gggtggagga
     3961 agctgacaac aatgaaaatc ttcaggagat atgcaaagca gaaaccctcg tgcaggcccg
     4021 aaagagaaag cgaaccagta tcgagaaccg agtgagaggc aacctggaga atttgttcct
     4081 gcagtgcccg aaacccacac tgcagcagat cagccacatc gcccagcagc ttgggctcga
     4141 gaaggatgtg gtccgagtgt ggttctgtaa ccggcgccag aagggcaagc gatcaagcag
     4201 cgactatgca caacgagagg attttgaggc tgctgggtct cctttctcag ggggaccagt
     4261 gtcctttcct ctggccccag ggccccattt tggtacccca ggctatggga gccctcactt
     4321 cactgcactg tactcctcgg tccctttccc tgagggggaa gcctttcccc ctgtctctgt
     4381 caccactctg ggctctccca tgcattcaaa ccgcgccaag cgcggctccg gcgccaccaa
     4441 cttctccctg ctgaagcaga tgtacaacat gatggagacg gagctgaagc cgccgggccc
     4501 gcagcaaact tcggggggcg gcggcggcaa ctccaccgcg gcggcggccg gcggcaacca
     4561 gaaaaacagc ccggaccgcg tcaagcggcc catgaatgcc ttcatggtgt ggtcccgcgg
     4621 gcagcggcgc aagatggccc aggagaaccc caagatgcac aactcggaga tcagcaagcg
     4681 cctgggcgcc gagtggaaac ttttgtcgga gacggagaag cggccgttca tcgacgaggc
     4741 taagcggctg cgagcgctgc acatgaagga gcacccggat tataaatacc ggccccggcg
     4801 gaaaaccaag acgctcatga agaaggataa gtacacgctg cccggcgggc tgctggcccc
     4861 cggcggcaat agcatggcga gcggggtcgg ggtgggcgcc ggcctgggcg cgggcgtgaa
     4921 ccagcgcatg gacagttacg cgcacatgaa cggctggagc aacggcagct acagcatgat
     4981 gcaggaccag ctgggctacc cgcagcaccc gggcctcaat gcgcacggcg cagcgcagat
     5041 gcagcccatg caccgctacg acgtgagcgc cctgcagtac aactccatga ccagctcgca
     5101 gacctacatg aacggctcgc ccacctacag catgtcctac tcgcagcagg gcacccctgg
     5161 catggctctt ggctccatgg gttcggtggt caagtccgag gccagctcca gcccccctgt
     5221 ggttacctct tcctcccact ccagggcgcc ctgccaggcc ggggacctcc gggacatgat
     5281 cagcatgtat ctccccggcg ccgaggtgcc ggaacccgcc gcccccagca gacttcacat
     5341 gtcccagcac taccagagcg gcccggtgcc cggcacggcc attaacggca cactgcccct
     5401 ctcacacatg cgcgccaagc gcggctccgg cgagggcaga ggaagtcttc taacatgcgg
     5461 tgacgtggag gagaatcccg gccctcctgc aggaatgagg cagccacctg gcgagtctga
     5521 catggctgtc agcgacgcgc tgctcccatc tttctccacg ttcgcgtctg gcccggcggg
     5581 aagggagaag acactgcgtc aagcaggtgc cccgaataac cgctggcggg aggagctctc
     5641 ccacatgaag cgacttcccc cagtgcttcc cggccgcccc tatgacctgg cggcggcgac
     5701 cgtggccaca gacctggaga gcggcggagc cggtgcggct tgcggcggta gcaacctggc
     5761 gcccctacct cggagagaga ccgaggagtt caacgatctc ctggacctgg actttattct
     5821 ctccaattcg ctgacccatc ctccggagtc agtggccgcc accgtgtcct cgtcagcgtc
     5881 agcctcctct tcgtcgtcgc cgtcgagcag cggccctgcc agcgcgccct ccacctgcag
     5941 cttcacctat ccgatccggg ccgggaacga cccgggcgtg gcgccgggcg gcacgggcgg
     6001 aggcctcctc tatggcaggg agtccgctcc ccctccgacg gctcccttca acctggcgga
     6061 catcaacgac gtgagcccct cgggcggctt cgtggccgag ctcctgcggc cagaattgga
     6121 cccggtgtac attccgccgc agcagccgca gccgccaggt ggcgggctga tgggcaagtt
     6181 cgtgctgaag gcgtcgctga gcgcccctgg cagcgagtac ggcagcccgt cggtcatcag
     6241 cgtcagcaaa ggcagccctg acggcagcca cccggtggtg gtggcgccct acaacggcgg
     6301 gccgccgcgc acgtgcccca agatcaagca ggaggcggtc tcttcgtgca cccacttggg
     6361 cgctggaccc cctctcagca atggccaccg gccggctgca cacgacttcc ccctggggcg
     6421 gcagctcccc agcaggacta ccccgaccct gggtcttgag gaagtgctga gcagcaggga
     6481 ctgtcaccct gccctgccgc ttcctcccgg cttccatccc cacccggggc ccaattaccc
     6541 atccttcctg cccgatcaga tgcagccgca agtcccgccg ctccattacc aagagctcat
     6601 gccacccggt tcctgcatgc cagaggagcc caagccaaag aggggaagac gatcgtggcc
     6661 ccggaaaagg accgccaccc acacttgtga ttacgcgggc tgcggcaaaa cctacacaaa
     6721 gagttcccat ctcaaggcac acctgcgaac ccacacaggt gagaaacctt accactgtga
     6781 ctgggacggc tgtggatgga aattcgcccg ctcagatgaa ctgaccaggc actaccgtaa
     6841 acacacgggg caccgcccgt tccagtgcca aaaatgcgac cgagcatttt ccaggtcgga
     6901 ccacctcgcc ttacacatga agaggcattt tggccggcca cgcgccaagc gcggctccgg
     6961 ccagtgcacc aactacgccc tgctgaagct ggccggcgac gtggagtcca accccggccc
     7021 cgcggccgca tttaaatcga tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     7081 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     7141 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     7201 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     7261 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     7321 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     7381 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     7441 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     7501 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     7561 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     7621 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     7681 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     7741 ggacgagctg tacaagtaat tcgaaccatg gcccaacttg tttattgcag cttataatgg
     7801 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     7861 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct cgaagttcct
     7921 attctctaga aagtatagga acttcccata gagcccaccg catccccagc atgcctgcta
     7981 ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc caccccccag aatagaatga
     8041 cacctactca gacaatgcga tgcaatttcc tcattttatt aggaaaggac agtgggagtg
     8101 gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg ctggcaacta
     8161 gaaggcacag ttagccctcc cacacataac cagagggcag caattcacga atcccaactg
     8221 ccgtcggctg tccatcactg tccttcacta tggctttgat cccaggatgc agatcgagaa
     8281 gcacctgtcg gcaccgtccg caggggctca agatgcccct gttctcattt ccgatcgcga
     8341 cgatacaagt caggttgcca gctgccgcag cagcagcagt gcccagcacc acgagttctg
     8401 cacaaggtcc cccagtaaaa tgatatacat tgacaccagt gaagatgcgg ccgtcgctag
     8461 agagagctgc gctggcgacg ctgtagtctt cagagatggg gatgctgttg attgtagccg
     8521 ttgctctttc aatgagggtg gattcttctt gagacaaagg cttggccatg gtggcggcag
     8581 ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga
     8641 ggcagaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
     8701 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     8761 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     8821 tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     8881 gcctggggac tttccacacc ctaactgaca cacattccac aggaagttcc tattctctag
     8941 aaagtatagg aacttcataa cttcgtatag catacattat acgaagttat ttaattaaag
     9001 ctgcaatcaa tcctgttcta aaaaccatca acttttaata agagatacat ctaacatagg
     9061 gcttacagtc atagcagcct agttataagt acaaaatagc caatatcaaa gttgttttgt
     9121 atgaatatag atacattgct atagagaaat agaagaacac ttggcacaca aacattgggt
     9181 tgcttcatta gcatttcttt ttactttaag caagatttct tgagggacat cttttctacg
     9241 ttggagtaaa aaatacgtta tttaaacttg aatttttttt tcagtaagtg aaacctatat
     9301 tattcagcca aatccttaag cttgagttaa ataattgaaa cagaaattta aaagttataa
     9361 ctaatggata ttcagacatt atacatttct tacccaagtg ttataatagg gacctactat
     9421 tttaagtgac acttaaacat ttatagttga cttatagacc ataaatttta atgatcataa
     9481 attttgattt acacaattat tgagttaaca agtcacaaag ttagtctatt tttataaagg
     9541 taaaaattag gaatcagatc aaaacattgt gaggtgcctt ctcagatgtt atacccatac
     9601 tttctattct taagtttaga aaataaataa aattcttact aaatctcttc tcactttatc
     9661 aaaaatattt tgcaggttgt cctgatagga aattgaggga ataatagctg ttcctttgga
     9721 cgaggatcat tggtgctatt cattccaaat gtgactttat gtaattaatt atcaacttca
     9781 aaccactgcc attggaatgg cgtaatcatg gtcatagctg tttcggctcg agcatggtca
     9841 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     9901 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     9961 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
    10021 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
    10081 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
    10141 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
    10201 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
    10261 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
    10321 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
    10381 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
    10441 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
    10501 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
    10561 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
    10621 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
    10681 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
    10741 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
    10801 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
    10861 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
    10921 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
    10981 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
    11041 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
    11101 ggcttaccat ctggccccag tgctgcaatg ataccgcgac tcccacgctc accggctcca
    11161 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
    11221 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
    11281 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
    11341 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
    11401 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
    11461 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
    11521 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
    11581 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
    11641 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
    11701 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
    11761 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
    11821 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
    11881 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
    11941 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
    12001 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
