LOCUS       sequence-442482-       12898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac transposon plasmid for doxycycline inducible expression of 
            TET1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12898)
  AUTHORS   Pierson Smela M, Adams J, Ma C, Breimann L, Widocki U, Shioda T, 
            Church GM
  TITLE     Induction of Meiosis from Human Pluripotent Stem Cells
  JOURNAL   bioRxiv 2024.05.31.596483
REFERENCE   2  (bases 1 to 12898)
  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: "bioRxiv 
            2024.05.31.596483"
COMMENT     Sequence Label: pPB-cT3G-cERP2-TET1
FEATURES             Location/Qualifiers
     source          1..12898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    17..35
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    53..71
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    89..107
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    125..143
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    161..179
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    197..215
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    233..251
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     278..302
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    368..392
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6813..6837)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6839..6865
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    7032..7166
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   7438..7472
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7483..7500)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7483..7499)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7492..7514)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7708..7727)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7829..7851
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7890..7908)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7976..8080
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8081..8149,8150..8941)
                     /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(8299..8318)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9112..9700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9601..9620
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9854..9871
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10077..10098
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10113..10118,10119..10136,10137..10143)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10151..10167
                     /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     10156..10178
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10175..10191
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10175..10191
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   10217..10279
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    10455..10536
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(11253..11309)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(11319..12062)
                     /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(12063..12086)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      12086..12095
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(12115..12326)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     enhancer        12327..12706
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atcacctcga gtttactccc tatcagtgat agagaacgta tgaagagttt actccctatc
       61 agtgatagag aacgtatgca gactttactc cctatcagtg atagagaacg tataaggagt
      121 ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa
      181 cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc
      241 agtgatagag aacgtataag cttgcctata taagcagagc tcgtttagtg aaccgtcaga
      301 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca
      361 gcctaccaca agtttgtaca aaaaagcagg cttagccacc atgtctcgat cccgccatgc
      421 aaggccttcc agattagtca ggaaggaaga tgtaaacaaa aaaaagaaaa acagccaact
      481 acgaaagaca accaagggag ccaacaaaaa tgtggcatca gtcaagactt taagccctgg
      541 aaaattaaag caattaattc aagaaagaga tgttaagaaa aaaacagaac ctaaaccacc
      601 cgtgccagtc agaagccttc tgacaagagc tggagcagca cgcatgaatt tggataggac
      661 tgaggttctt tttcagaacc cagagtcctt aacctgcaat gggtttacaa tggcgctacg
      721 aagcacctct cttagcaggc gactctccca acccccactg gtcgtagcca aatccaaaaa
      781 ggttccactt tctaagggtt tagaaaagca acatgattgt gattataaga tactccctgc
      841 tttgggagta aagcactcag aaaatgattc ggttccaatg caagacaccc aagtccttcc
      901 tgatatagag actctaattg gtgtacaaaa tccctcttta cttaaaggta agagccaaga
      961 gacaactcag ttttggtccc aaagagttga ggattccaag atcaatatcc ctacccacag
     1021 tggccctgca gctgagatcc ttcctgggcc actggaaggg acacgctgtg gtgaaggact
     1081 attctctgaa gagacattga atgataccag tggttcccca aaaatgtttg ctcaggacac
     1141 agtgtgtgct ccttttcccc aaagagcaac ccccaaagtt acctctcaag gaaaccccag
     1201 cattcagtta gaagagttgg gttcacgagt agaatctctt aagttatctg attcttacct
     1261 ggatcccatt aaaagtgaac atgattgcta ccccacctcc agtcttaata aggttatacc
     1321 tgacttgaac cttagaaact gcttggctct tggtgggtct acgtctccta cctctgtaat
     1381 aaaattcctc ttggcaggct caaaacaagc gacccttggt gctaaaccag atcatcaaga
     1441 ggccttcgaa gctactgcaa atcaacagga agtttctgat accacctctt tcctaggaca
     1501 ggcctttggt gctatcccac atcaatggga acttcctggt gctgacccag ttcatggtga
     1561 ggccctgggt gagaccccag atctaccaga gattcctggt gctattccag tccaaggaga
     1621 ggtctttggt actattttag accaacaaga aactcttggt atgagtggga gtgttgtccc
     1681 agacttgcct gtcttccttc ctgttcctcc aaatccaatt gctaccttta atgctccttc
     1741 caaatggcct gagccccaaa gcactgtctc atatggactt gcagtccagg gtgctataca
     1801 gattttgcct ttgggctcag gacacactcc tcaatcatca tcaaactcag agaaaaattc
     1861 attacctcca gtaatggcta taagcaatgt agaaaatgag aagcaggttc atataagctt
     1921 cctgccagct aacactcagg ggttcccatt agcccctgag agaggactct tccatgcttc
     1981 actgggtata gcccaactct ctcaggctgg tcctagcaaa tcagacagag ggagctccca
     2041 ggtcagtgta accagcacag ttcatgttgt caacaccaca gtggtgacta tgccagtgcc
     2101 aatggtcagt acctcctctt cttcctatac cactttgcta ccgactttgg aaaagaagaa
     2161 aagaaagcga tgtggggtct gtgaaccctg ccagcagaag accaactgtg gtgaatgcac
     2221 ttactgcaag aacagaaaga acagccatca gatctgtaag aaaagaaaat gtgaggagct
     2281 gaaaaagaaa ccatctgttg ttgtgcctct ggaggttata aaggaaaaca agaggcccca
     2341 gagggaaaag aagcccaaag ttttaaaggc agattttgac aacaaaccag taaatggccc
     2401 caagtcagaa tccatggact acagtagatg tggtcatggg gaagaacaaa aattggaatt
     2461 gaacccacat actgttgaaa atgtaactaa aaatgaagac agcatgacag gcatcgaggt
     2521 ggagaagtgg acacaaaaca agaaatcaca gttaactgat cacgtgaaag gagattttag
     2581 tgctaatgtc ccagaagctg aaaaatcgaa aaactctgaa gttgacaaga aacgaaccaa
     2641 atctccaaaa ttgtttgtac aaaccgtaag aaatggcatt aaacatgtac actgtttacc
     2701 agctgaaaca aatgtttcat ttaaaaaatt caatattgaa gaattcggca agacattgga
     2761 aaacaattct tataaattcc taaaagacac tgcaaaccat aaaaacgcta tgagctctgt
     2821 tgctactgat atgagttgtg atcatctcaa ggggagaagt aacgttttag tattccagca
     2881 gcctggcttt aactgcagtt ccattccaca ttcttcacac tccatcataa atcatcatgc
     2941 tagtatacac aatgaaggtg atcaaccaaa aactcctgag aatataccaa gtaaagaacc
     3001 aaaagatgga tctcccgttc aaccaagtct cttatcgtta atgaaagata ggagattaac
     3061 attggagcaa gtggtagcca tagaggccct gactcaactc tcagaagccc catcagagaa
     3121 ttcctcccca tcaaagtcag agaaggatga ggaatcagag cagagaacag ccagtttgct
     3181 taatagctgc aaagctatcc tctacactgt aagaaaagac ctccaagacc caaacttaca
     3241 gggagagcca ccaaaactta atcactgtcc atctttggaa aaacaaagtt catgcaacac
     3301 ggtggttttc aatgggcaaa ctactaccct ttccaactca catatcaact cagctactaa
     3361 ccaagcatcc acaaagtcac atgaatattc aaaagtcaca aattcattat ctctttttat
     3421 accaaaatca aattcatcca agattgacac caataaaagt attgctcaag ggataattac
     3481 tcttgacaat tgttccaatg atttgcatca gttgccacca agaaataatg aagtggagta
     3541 ttgcaaccag ttactggaca gcagcaaaaa attggactca gatgatctat catgtcagga
     3601 tgcaacccat acccaaattg aggaagatgt tgcaacacag ttgacacaac ttgcttcgat
     3661 aattaagatc aattatataa aaccagagga caaaaaagtt gaaagtacac caacaagcct
     3721 tgtcacatgt aatgtacagc aaaaatacaa tcaggagaag ggcacaatgc aacagaaacc
     3781 accttcaagt gtacacaata atcatggttc atcattaaca aaacaaaaga acccaaccca
     3841 gaaaaagaca aaatccaccc catcaagaga tcggcggaaa aagaagccca cagttgtaag
     3901 ttatcaagaa aatgatcggc agaagtggga aaagttgtcc tatatgtatg gcacaatatg
     3961 cgacatttgg atagcatcga aatttcaaaa ttttgggcaa ttttgtccac atgattttcc
     4021 tactgtattt gggaaaattt cttcctcgac caaaatatgg aaaccactgg ctcaaacgag
     4081 gtccattatg caacccaaaa cagtatttcc accactcact cagataaaat tacagagata
     4141 tcctgaatca gcagaggaaa aggtgaaggt tgaaccattg gattcactca gcttatttca
     4201 tcttaaaacg gaatccaacg ggaaggcatt cactgataaa gcttataatt ctcaggtaca
     4261 gttaacggtg aatgccaatc agaaagccca tcctttgacc cagccctcct ctccacctaa
     4321 ccagtgtgct aacgtgatgg caggcgatga ccaaatacgg tttcagcagg ttgttaagga
     4381 gcaactcatg catcagagac tgccaacatt gcctggtatc tctcatgaaa cacccttacc
     4441 ggagtcagca ctaactctca ggaatgtaaa tgtagtgtgt tcaggtggaa ttacagtggt
     4501 ttctaccaaa agtgaagagg aagtctgttc atccagtttt ggaacatcag aattttccac
     4561 agtggacagt gcacagaaaa attttaatga ttatgccatg aacttcttta ctaaccctac
     4621 aaaaaaccta gtgtctataa ctaaagattc tgaactgccc acctgcagct gtcttgatcg
     4681 agttatacaa aaagacaaag gcccatatta tacacacctt ggggcaggac caagtgttgc
     4741 tgctgtcagg gaaatcatgg agaataggta tggtcaaaaa ggaaacgcaa taaggataga
     4801 aatagtagtg tacaccggta aagaagggaa aagctctcat gggtgtccaa ttgctaagtg
     4861 ggttttaaga agaagcagtg atgaagaaaa agttctttgt ttggtccggc agcgtacagg
     4921 ccaccactgt ccaactgctg tgatggtggt gctcatcatg gtgtgggatg gcatccctct
     4981 tccaatggcc gaccggctat acacagagct cacagagaat ctaaagtcat acaatgggca
     5041 ccctaccgac agaagatgca ccctcaatga aaatcgtacc tgtacatgtc aaggaattga
     5101 tccagagact tgtggagctt cattctcttt tggctgttca tggagtatgt actttaatgg
     5161 ctgtaagttt ggtagaagcc caagccccag aagatttaga attgatccaa gctctccctt
     5221 acatgaaaaa aaccttgaag ataacttaca gagtttggct acacgattag ctccaattta
     5281 taagcagtat gctccagtag cttaccaaaa tcaggtggaa tatgaaaatg ttgcccgaga
     5341 atgtcggctt ggcagcaagg aaggtcgtcc cttctctggg gtcactgctt gcctggactt
     5401 ctgtgctcat ccccacaggg acattcacaa catgaataat ggaagcactg tggtttgtac
     5461 cttaactcga gaagataacc gctctttggg tgttattcct caagatgagc agctccatgt
     5521 gctacctctt tataagcttt cagacacaga tgagtttggc tccaaggaag gaatggaagc
     5581 caagatcaaa tctggggcca tcgaggtcct ggcaccccgc cgcaaaaaaa gaacgtgttt
     5641 cactcagcct gttccccgtt ctggaaagaa gagggctgcg atgatgacag aggttcttgc
     5701 acataagata agggcagtgg aaaagaaacc tattccccga atcaagcgga agaataactc
     5761 aacaacaaca aacaacagta agccttcgtc actgccaacc ttagggagta acactgagac
     5821 cgtgcaacct gaagtaaaaa gtgaaaccga accccatttt atcttaaaaa gttcagacaa
     5881 cactaaaact tattcgctga tgccatccgc tcctcaccca gtgaaagagg catctccagg
     5941 cttctcctgg tccccgaaga ctgcttcagc cacaccagct ccactgaaga atgacgcaac
     6001 agcctcatgc gggttttcag aaagaagcag cactccccac tgtacgatgc cttcgggaag
     6061 actcagtggt gccaatgcag ctgctgctga tggccctggc atttcacagc ttggcgaagt
     6121 ggctcctctc cccaccctgt ctgctcctgt gatggagccc ctcattaatt ctgagccttc
     6181 cactggtgtg actgagccgc taacgcctca tcagccaaac caccagccct ccttcctcac
     6241 ctctcctcaa gaccttgcct cttctccaat ggaagaagat gagcagcatt ctgaagcaga
     6301 tgagcctcca tcagacgaac ccctatctga tgaccccctg tcacctgctg aggagaaatt
     6361 gccccacatt gatgagtatt ggtcagacag tgagcacatc tttttggatg caaatattgg
     6421 tggggtggcc atcgcacctg ctcacggctc ggttttgatt gagtgtgccc ggcgagagct
     6481 gcacgctacc actcctgttg agcaccccaa ccgtaatcat ccaacccgcc tctcccttgt
     6541 cttttaccag cacaaaaacc taaataagcc ccaacatggt tttgaactaa acaagattaa
     6601 gtttgaggct aaagaagcta agaataagaa aatgaaggcc tcagagcaaa aagaccaggc
     6661 agctaatgaa ggtccagaac agtcctctga agtaaatgaa ttgaaccaaa ttccttctca
     6721 taaagcatta acattaaccc atgacaatgt tgtcaccgtg tccccttatg ctctcacaca
     6781 cgttgcgggg ccctataacc attgggtctg atacccagct ttcttgtaca aagtggtata
     6841 cccatacgat gttccagatt acgcttagta atgagaaata ctcagaagat gtcactcacc
     6901 atagtccgca gtgaaagata aatgatcggt attttgcccg tgattctgtt ttagagctag
     6961 aaatagcaag taggccactg agcaagaact atgaatgaga tctttagaca tattgcggcc
     7021 gctttctccg cgatccagac atgataagat acattgatga gtttggacaa accaaaacta
     7081 gaatgcagtg aaaaaaatgc cttatttgtg aaatttgtga tgctattgcc ttatttgtaa
     7141 ccattataag ctgcaataaa caagttatct ataacaagaa aatatatata taataagtta
     7201 tcacgtaagt agaacatgaa ataacaatat aattatcgta tgagttaaat cttaaaagtc
     7261 acgtaaaaga taatcatgcg tcattttgac tcacgcggtc gttatagttc aaaatcagtg
     7321 acacttaccg cattgacaag cacgcctcac gggagctcca agcggcgact gagatgtcct
     7381 aaatgcacag cgacggattc gcgctattta gaaagagaga gcaatatttc aagaatgcat
     7441 gcgtcaattt tacgcagact atctttctag ggttaagaat tcactggccg tcgttttaca
     7501 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     7561 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     7621 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     7681 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     7741 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     7801 cgcttacaga caagctgtga ccatgtctcc gggagctgca tgtgtcagag gttttcaccg
     7861 tcatcaccga aacacgcgag ttcgaaaggg cctcgtgata cgcctatttt tataggttaa
     7921 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     7981 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     8041 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     8101 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     8161 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     8221 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     8281 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     8341 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     8401 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     8461 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     8521 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     8581 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     8641 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     8701 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     8761 gtttattgct gataaatctg gagccggtga gcgtggttca cgcggtatca ttgcagcact
     8821 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     8881 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     8941 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     9001 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     9061 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     9121 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     9181 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     9241 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     9301 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     9361 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     9421 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     9481 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     9541 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     9601 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     9661 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     9721 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     9781 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     9841 aacgaccgag cgcagcgagt cagtgagcga ggaagcggat gagcgcccaa tacgcaaacc
     9901 gcctctcacc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     9961 gaaagcgggc agtgagcgca acgcaaaccg cctctccccg cgcgttggcc gattcattaa
    10021 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
    10081 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
    10141 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
    10201 gccaaggtcg acttaaccct agaaagataa tcatattgtg acgtacgtta aagataatca
    10261 tgcgtaaaat tgacgcatgt gttttatcgg tctgtatatc gaggtttatt tattaatttg
    10321 aatagatatt aagttttatt atatttacac ttacatacta ataataaatt caacaaacaa
    10381 tttatttatg tttatttatt tattaaaaaa aaacaaaaac tcaaaatttc ttctataaag
    10441 taacaaaact tttagcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
    10501 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
    10561 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
    10621 tgtggtatgg ctgattatga tcctctggag attcatgcgc caggttttct ggtcatgcac
    10681 cacgttcgtg ggccctcagg aacctccacg tcagcggtaa ctgtgaagcc cagtcgctcg
    10741 taaaaaggaa gattccgggg agcactggtt tcgaggaacg ccggcacgcc agcccgttcc
    10801 gcggcctcaa caccaggaag aacgacagcg cttcccaaac ctttaccttg gtgatctggg
    10861 ctgacaccga cagtggcaag aaaccacgca ggctcctttg gtctatgagg agccagcaag
    10921 ccctccattt gctgttgtgc cgccaaccta ctccctgaca gctcggccat ccgcggacct
    10981 atttccgcga acactgcgcc ggcttcgacg ctttccggag tagtccagac cgcaactgcg
    11041 gcaccatcgt ccgcaaccca aactttacca atgtccagac ccacgcgcgt aagaaacagc
    11101 tcttgcaact cagttaccct ctctatgtgt ctgtccggat ctacggtatg gcgggtcgct
    11161 ggataatctg caaatgctgc ggcaagtgtt cggacagccc gtggtacatc gtcacgggta
    11221 gcgagcctaa cagtgggctt atattccgtc atggggccgg ggttctcctc cacgtcgccg
    11281 gcctgcttca gcaggctgaa gttggtggcg ccgctgcccc cggggagcat gtcaaggtca
    11341 aaatcgtcaa gagcgtcagc aggcagcata tcaaggtcaa agtcgtcaag ggcatcggct
    11401 gggagcatgt ctaagtcaaa atcgtcaagg gcgtcggtcg gcccgccgct ttcgcacttt
    11461 agctgtttct ccaggccaca tatgattagt tccaggccga aaaggaaggc aggttcggct
    11521 ccctgccggt cgaacagctc aattgcttgt ttcagaagtg ggggcataga atcggtggta
    11581 ggtgtctctc tttcctcttt tgctacttga tgctcctgtt cctccaatac gcagcccagt
    11641 gtaaagtggc ccacggcgga cagagcgtac agtgcgttct ccagggagaa gccttgctga
    11701 cacaggaacg cgagctgatt ttccagggtt tcgtactgtt tctctgttgg gcgggtgccg
    11761 agatgcactt tagccccgtc acgatgtgag aggagagcac agcggtatga cttggcgttg
    11821 ttccgcagaa agtcttgcca tgactcgcct tccagggggc aggagtgggt atgatgcctg
    11881 tccagcatct cgattggcag ggcatcgagc agggcccgct tgttcttcac gtgccagtac
    11941 agggtaggct gctcaactcc cagcttttga gcgagtttcc ttgtcgtcag gccttcgata
    12001 ccgactccat tgagtaattc cagagcagag tttatgactt tgctcttgtc cagtctagac
    12061 atcttatcgt catcgtcttt gtaatccatg gtggcggatc ccgcgtcacg acacctgtgt
    12121 tctggcggca aacccgttgc gaaaaagaac gttcacggcg actactgcac ttatatacgg
    12181 ttctccccca ccctcgggaa aaaggcggag ccagtacacg acatcacttt cccagtttac
    12241 cccgcgccac cttctctagg caccggttca attgccgacc cctcccccca acttctcggg
    12301 cactgtgggc catgtgcgct ctgccccatg gtaatagcca tgactaatac gtagatgtac
    12361 tgccaagtag gaaagtgcca taaggtcatg tactgggcat aatgccaggc gggccattta
    12421 ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag
    12481 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtgcc tattggcgtt
    12541 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
    12601 ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat
    12661 gaccccgtaa ttgattacta ttaataacta gtcaataatc aatgtcataa gctctaatcg
    12721 cataatcgag ctaagagcga ttaatatgct tagctagctt agctaagcta gatttctcta
    12781 ttctcccaca atatatagca gctattagct tagcagtccg aatacgatcg tccacgatcc
    12841 ataagcgtaa cacatttgag tagcgactga cgggcaccgg agccactcga gtggaatt
//
