PB_iETV2_P2A_GFP_Puro

LOCUS       PB_iETV2_P2A_GFP_Puro         9423 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  PiggyBac transposon construct with doxycycline inducible ETV2 + GFP 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    PB_iETV2_P2A_GFP_Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9423)
  TITLE     Programmatic Introduction of Parenchymal Cell Types in Blood Vessel 
            Organoids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9423)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9423
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     regulatory      13..22
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             812..823
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1138..1194
                     /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             1195..1914
                     /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             1195..1911
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1240..1261)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1501..1520)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1848..1869
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1951..1970)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1995..2043
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        2274..2485
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2498..2766
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2800..3543
                     /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             3544..3597
                     /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             3598..4197
                     /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(3598..3617)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4094..4114
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4230..4351
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4267..4286)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4321..4340
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   5114..5148
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5639..5655)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5639..5655)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5652..5674)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5663..5679
                     /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(5687..5717)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5732..5753
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5870..5887)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6041..6629)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6121..6140)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6800..7660)
                     /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     7423..7442
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7661..7765)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7791..8246)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7928..7949)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8140..8159
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8373..8395
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8387..8404
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8388..8404
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   8534..8596
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        9045..9423
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    9053..9071
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9089..9107
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9125..9143
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9161..9179
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9197..9215
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9233..9251
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9269..9287
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     9332..9356
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 ggtctagagc tagccaccat ggatctttgg aactgggatg aagcttcccc tcaagaagtt
       61 ccccccggaa ataaactcgc ggggcttgga agactccctc gccttccgca acgcgtctgg
      121 ggcggatgcc ctggtggagc ctcagcggac ccaaaccctt tgtctccagc ggagggggca
      181 aagttgggtt tctgcttccc ggatcttgct ttgcaaggcg atactccaac ggcgacggca
      241 gagacctgtt ggaaaggcac cagtagctcc ctggccagct ttccgcagct cgattggggg
      301 tcagcccttc tccatcccga agttccctgg ggggcggaac ccgactccca agcccttccc
      361 tggagtggtg attggacaga tatggcatgc acagcctggg acagttggtc cggggcgtca
      421 cagacattgg gaccagcccc acttggaccg gggcctatcc ccgcagcagg aagcgaagga
      481 gctgctggtc agaactgtgt gcccgtggct ggtgaggcta ccagttggtc cagggcccag
      541 gcagcaggca gtaacaccag ctgggattgc tcagtggggc ctgacgggga tacttattgg
      601 ggctctggtc ttggtggaga accgagaacg gactgtacga taagttgggg cggtccagct
      661 gggcctgatt gtactacgtc atggaatcct ggcttgcacg ccggcggcac gacaagcctt
      721 aagagatatc aaagttcagc ccttacagtt tgctcagaac cttccccgca aagtgaccga
      781 gcgtcactgg cgcgatgtcc taaaactaat catcgagggc cgatccagtt gtggcagttt
      841 ttgcttgaac tccttcacga tggcgcgagg agcagttgca tcagatggac cggtaacagc
      901 agggagttcc aattgtgtga ccccaaggaa gtggctcgac tgtggggtga gcgcaaacgg
      961 aagcctggta tgaattacga aaagttgagt aggggtttgc gatattacta taggcgcgac
     1021 atcgttcgaa agtccggtgg tcgaaagtac acatacagat tcggcggtcg cgtaccatct
     1081 cttgcatacc ctgattgcgc aggcgggggt aggggtgcgg aaacacaagg atccggcgca
     1141 acaaacttct ctctgctgaa acaagccgga gatgtcgaag agaatcctgg accgatggtg
     1201 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1261 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1321 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1381 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     1441 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1501 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1561 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1621 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1681 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1741 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1801 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1861 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaaccggt
     1921 tagtaatgag tttaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     1981 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg
     2041 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     2101 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     2161 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgaa
     2221 atcggatcca gctgaatcta agtggatctg cgatcgctcc ggtgcccgtc agtgggcaga
     2281 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc
     2341 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     2401 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     2461 caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg
     2521 cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc
     2581 cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag
     2641 accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct
     2701 ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta
     2761 cagatctaag ctgtgaccgg cgcctactct agagccgcca tgtctagact ggacaagagc
     2821 aaagtcataa actctgctct ggaattactc aatggagtcg gtatcgaagg cctgacgaca
     2881 aggaaactcg ctcaaaagct gggagttgag cagcctaccc tgtactggca cgtgaagaac
     2941 aagcgggccc tgctcgatgc cctgccaatc gagatgctgg acaggcatca tacccactcc
     3001 tgccccctgg aaggcgagtc atggcaagac tttctgcgga acaacgccaa gtcataccgc
     3061 tgtgctctcc tctcacatcg cgacggggct aaagtgcatc tcggcacccg cccaacagag
     3121 aaacagtacg aaaccctgga aaatcagctc gcgttcctgt gtcagcaagg cttctccctg
     3181 gagaacgcac tgtacgctct gtccgccgtg ggccacttta cactgggctg cgtattggag
     3241 gaacaggagc atcaagtagc aaaagaggaa agagagacac ctaccaccga ttctatgccc
     3301 ccacttctga aacaagcaat tgagctgttc gaccggcagg gagccgaacc tgccttcctt
     3361 ttcggcctgg aactaatcat atgtggcctg gagaaacagc taaagtgcga aagcggcggg
     3421 ccgaccgacg cccttgacga ttttgactta gacatgctcc cagccgatgc ccttgacgac
     3481 tttgaccttg atatgctgcc tgctgacgct cttgacgatt ttgaccttga catgctcccc
     3541 ggggagggcc gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctatg
     3601 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     3661 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     3721 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     3781 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     3841 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     3901 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     3961 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     4021 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     4081 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     4141 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaaat
     4201 caacctctgg attacaaaat ttgtgggtaa acttgtttat tgcagcttat aatggttaca
     4261 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4321 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gaattgactc aaatgatgtc
     4381 aattagtcta tcagaagctc atctggtctc ccttccgggg gacaagacat ccctgtttaa
     4441 tatttaaaca gcagtgttcc caaactgggt tcttatatcc cttgctctgg tcaaccaggt
     4501 tgcagggttt cctgtcctca caggaacgaa gtccctaaag aaacagtggc agccaggttt
     4561 agccccggaa ttgactggat tcctttttta gggcccattg gtatggcttt ttccccgtat
     4621 ccccccaggt gtctgcaggc tcaaagagca gcgagaagcg ttcagaggaa agcgatcccg
     4681 tgccaccttc cccgtgcccg ggctgtcccc gcacgctgcc ggctcgggga tgcgggggga
     4741 gcgccggacc ggagcggagc cccgggcggc tcgctgctgc cccctagcgg gggagggacg
     4801 taattacatc cctgggggct ttgggggggg gctgtccctg atatctataa caagaaaata
     4861 tatatataat aagttatcac gtaagtagaa catgaaataa caatataatt atcgtatgag
     4921 ttaaatctta aaagtcacgt aaaagataat catgcgtcat tttgactcac gcggtcgtta
     4981 tagttcaaaa tcagtgacac ttaccgcatt gacaagcacg cctcacggga gctccaagcg
     5041 gcgactgaga tgtcctaaat gcacagcgac ggattcgcgc tatttagaaa gagagagcaa
     5101 tatttcaaga atgcatgcgt caattttacg cagactatct ttctagggtt aatctagctg
     5161 catcaggatc atatcgtcgg gtcttttttc cggctcagtc atcgcccaag ctggcgctat
     5221 ctgggcatcg gggaggaaga agcccgtgcc ttttcccgcg aggttgaagc ggcatggaaa
     5281 gagtttgccg aggatgactg ctgctgcatt gacgttgagc gaaaacgcac gtttaccatg
     5341 atgattcggg aaggtgtggc catgcacgcc tttaacggtg aactgttcgt tcaggccacc
     5401 tgggatacca gttcgtcgcg gcttttccgg acacagttcc ggatggtcag cccgaagcgc
     5461 atcagcaacc cgaacaatac cggcgacagc cggaactgcc gtgccggtgt gcagattaat
     5521 gacagcggtg cggcgctggg atattacgtc agcgaggacg ggtatcctgg ctggatgccg
     5581 cagaaatgga catggatacc ccgtgagtta cccggcgggc gcgcttggcg taatcatggt
     5641 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5701 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5761 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5821 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5881 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5941 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6001 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6061 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6121 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6181 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6241 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6301 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6361 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6421 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6481 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6541 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6601 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6661 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6721 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6781 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6841 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6901 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6961 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7021 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7081 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7141 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7201 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7261 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7321 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7381 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7441 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7501 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7561 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7621 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7681 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7741 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag
     7801 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     7861 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     7921 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     7981 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     8041 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     8101 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     8161 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     8221 gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga
     8281 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     8341 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     8401 cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg gaggacgggc agactcgcgg
     8461 tgcaaatgtg ttttacagcg tgatggagca gatgaagatg ctcgacacgc tgcagaacac
     8521 gcagctagat taaccctaga aagataatca tattgtgacg tacgttaaag ataatcatgt
     8581 gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag gtttatttat taatttgaat
     8641 agatattaag ttttattata tttacactta catactaata ataaattcaa caaacaattt
     8701 atttatgttt atttatttat taaaaaaaac aaaaactcaa aatttcttct ataaagtaac
     8761 aaaactttta tgagggacag ccccccccca aagcccccag ggatgtaatt acgtccctcc
     8821 cccgctaggg ggcagcagcg agccgcccgg ggctccgctc cggtccggcg ctccccccgc
     8881 atccccgagc cggcagcgtg cggggacagc ccgggcacgg ggaaggtggc acgggatcgc
     8941 tttcctctga acgcttctcg ctgctctttg agcctgcaga cacctggggg gatacgggga
     9001 aaaggcctcc acggccacta gtctttcgtc ttcaagaatt cctcgagttt actccctatc
     9061 agtgatagag aacgtatgaa gagtttactc cctatcagtg atagagaacg tatgcagact
     9121 ttactcccta tcagtgatag agaacgtata aggagtttac tccctatcag tgatagagaa
     9181 cgtatgacca gtttactccc tatcagtgat agagaacgta tctacagttt actccctatc
     9241 agtgatagag aacgtatatc cagtttactc cctatcagtg atagagaacg tataagcttt
     9301 aggcgtgtac ggtgggcgcc tataaaagca gagctcgttt agtgaaccgt cagatcgcct
     9361 ggagcaattc cacaacactt ttgtcttata ccaactttcc gtaccacttc ctaccctcgt
     9421 aaa
//