LOCUS       sequence-332421-       10050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac transposon construct with doxycycline inducible ASCL1, DLX2
            + mCherry expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10050)
  AUTHORS   Dailamy A, Parekh U, Katrekar D, Kumar A, McDonald D, Moreno A, 
            Bagheri P, Ng TN, Mali P
  TITLE     Programmatic introduction of parenchymal cell types into blood 
            vessel organoids.
  JOURNAL   Stem Cell Reports. 2021 Sep 8. pii: S2213-6711(21)00432-X. doi: 
            10.1016/j.stemcr.2021.08.014.
  PUBMED    34559998
REFERENCE   2  (bases 1 to 10050)
  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 Cell 
            Reports. 2021 Sep 8. pii: S2213-6711(21)00432-X. doi: 
            10.1016/j.stemcr.2021.08.014."
COMMENT     Sequence Label: PB_iASCL1_P2A_DLX2_T2A_mCherry_Puro
FEATURES             Location/Qualifiers
     source          1..10050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             330..350
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             390..443
                     /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             444..1154
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(589..607)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     857..876
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1191..1210)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1235..1283
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        1514..1725
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1738..2006
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2040..2783
                     /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             2784..2837
                     /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             2838..3437
                     /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(2838..2857)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3334..3354
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3470..3591
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3507..3526)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3561..3580
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4354..4388
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4879..4895)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4879..4895)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4892..4914)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4903..4919
                     /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(4927..4933,4934..4951,4952..4957))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4972..4993
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5110..5127)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5281..5869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5361..5380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6040..6831,6832..6900))
                     /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     6663..6682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6901..7005)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7031..7486)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7168..7189)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7380..7399
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7613..7635
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7627..7644
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7628..7644
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   7774..7836
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        8285..8663
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    8293..8311
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8329..8347
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8365..8383
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8401..8419
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8437..8455
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8473..8491
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8509..8527
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     8572..8596
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      8676..8685
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             9399..9455
                     /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"
ORIGIN
        1 ggttccagaa ccgccggtcc aagttcaaga agatgtggaa aagtggtgag atcccctcgg
       61 agcagcaccc tggggccagc gcttctccac cttgtgcttc gccgccagtc tcagcgccgg
      121 cctcctggga ctttggtgtg ccgcagcgga tggcgggcgg cggtggtccg ggcagtggcg
      181 gcagcggcgc cggcagctcg ggctccagcc cgagcagcgc ggcctcggct tttctgggca
      241 actacccctg gtaccaccag acctcgggat ccgcctcaca cctgcaggcc acggcgccgc
      301 tgctgcaccc cactcagacc ccgcagccgc atcaccacca ccaccatcac ggcggcgggg
      361 gcgccccggt gagcgcgggg acgattttcg aaggtagagg ctcactgtta acgtgtggcg
      421 atgtagaaga gaatccagga cctatggtga gcaagggcga ggaggataac atggccatca
      481 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
      541 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
      601 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
      661 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
      721 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
      781 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
      841 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
      901 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
      961 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     1021 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     1081 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     1141 agctgtacaa gtaaaccggt tagtaatgag tttaaacggg ggaggctaac tgaaacacgg
     1201 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     1261 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     1321 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     1381 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     1441 tgccatagca gatctgcgaa atcggatcca gctgaatcta agtggatctg cgatcgctcc
     1501 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     1561 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     1621 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     1681 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg
     1741 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg
     1801 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta
     1861 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga
     1921 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg
     1981 ttttctgttc tgcgccgtta cagatctaag ctgtgaccgg cgcctactct agagccgcca
     2041 tgtctagact ggacaagagc aaagtcataa actctgctct ggaattactc aatggagtcg
     2101 gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct gggagttgag cagcctaccc
     2161 tgtactggca cgtgaagaac aagcgggccc tgctcgatgc cctgccaatc gagatgctgg
     2221 acaggcatca tacccactcc tgccccctgg aaggcgagtc atggcaagac tttctgcgga
     2281 acaacgccaa gtcataccgc tgtgctctcc tctcacatcg cgacggggct aaagtgcatc
     2341 tcggcacccg cccaacagag aaacagtacg aaaccctgga aaatcagctc gcgttcctgt
     2401 gtcagcaagg cttctccctg gagaacgcac tgtacgctct gtccgccgtg ggccacttta
     2461 cactgggctg cgtattggag gaacaggagc atcaagtagc aaaagaggaa agagagacac
     2521 ctaccaccga ttctatgccc ccacttctga aacaagcaat tgagctgttc gaccggcagg
     2581 gagccgaacc tgccttcctt ttcggcctgg aactaatcat atgtggcctg gagaaacagc
     2641 taaagtgcga aagcggcggg ccgaccgacg cccttgacga ttttgactta gacatgctcc
     2701 cagccgatgc ccttgacgac tttgaccttg atatgctgcc tgctgacgct cttgacgatt
     2761 ttgaccttga catgctcccc ggggagggcc gaggaagtct tctaacatgc ggtgacgtgg
     2821 aggagaatcc cggccctatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     2881 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     2941 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     3001 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     3061 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     3121 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     3181 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     3241 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     3301 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     3361 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     3421 gcaagcccgg tgcctgaaat caacctctgg attacaaaat ttgtgggtaa acttgtttat
     3481 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3541 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3601 gaattgactc aaatgatgtc aattagtcta tcagaagctc atctggtctc ccttccgggg
     3661 gacaagacat ccctgtttaa tatttaaaca gcagtgttcc caaactgggt tcttatatcc
     3721 cttgctctgg tcaaccaggt tgcagggttt cctgtcctca caggaacgaa gtccctaaag
     3781 aaacagtggc agccaggttt agccccggaa ttgactggat tcctttttta gggcccattg
     3841 gtatggcttt ttccccgtat ccccccaggt gtctgcaggc tcaaagagca gcgagaagcg
     3901 ttcagaggaa agcgatcccg tgccaccttc cccgtgcccg ggctgtcccc gcacgctgcc
     3961 ggctcgggga tgcgggggga gcgccggacc ggagcggagc cccgggcggc tcgctgctgc
     4021 cccctagcgg gggagggacg taattacatc cctgggggct ttgggggggg gctgtccctg
     4081 atatctataa caagaaaata tatatataat aagttatcac gtaagtagaa catgaaataa
     4141 caatataatt atcgtatgag ttaaatctta aaagtcacgt aaaagataat catgcgtcat
     4201 tttgactcac gcggtcgtta tagttcaaaa tcagtgacac ttaccgcatt gacaagcacg
     4261 cctcacggga gctccaagcg gcgactgaga tgtcctaaat gcacagcgac ggattcgcgc
     4321 tatttagaaa gagagagcaa tatttcaaga atgcatgcgt caattttacg cagactatct
     4381 ttctagggtt aatctagctg catcaggatc atatcgtcgg gtcttttttc cggctcagtc
     4441 atcgcccaag ctggcgctat ctgggcatcg gggaggaaga agcccgtgcc ttttcccgcg
     4501 aggttgaagc ggcatggaaa gagtttgccg aggatgactg ctgctgcatt gacgttgagc
     4561 gaaaacgcac gtttaccatg atgattcggg aaggtgtggc catgcacgcc tttaacggtg
     4621 aactgttcgt tcaggccacc tgggatacca gttcgtcgcg gcttttccgg acacagttcc
     4681 ggatggtcag cccgaagcgc atcagcaacc cgaacaatac cggcgacagc cggaactgcc
     4741 gtgccggtgt gcagattaat gacagcggtg cggcgctggg atattacgtc agcgaggacg
     4801 ggtatcctgg ctggatgccg cagaaatgga catggatacc ccgtgagtta cccggcgggc
     4861 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4921 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4981 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5041 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5101 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5161 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5221 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5281 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5341 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5401 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5461 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5521 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5581 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5641 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5701 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5761 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5821 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5881 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5941 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6001 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6061 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6121 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6181 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6241 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6301 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6361 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6421 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6481 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6541 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6601 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6661 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6721 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6781 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6841 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6901 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6961 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7021 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     7081 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     7141 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     7201 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     7261 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     7321 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     7381 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     7441 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
     7501 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     7561 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     7621 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
     7681 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
     7741 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
     7801 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
     7861 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     7921 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     7981 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     8041 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     8101 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     8161 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     8221 cacctggggg gatacgggga aaaggcctcc acggccacta gtctttcgtc ttcaagaatt
     8281 cctcgagttt actccctatc agtgatagag aacgtatgaa gagtttactc cctatcagtg
     8341 atagagaacg tatgcagact ttactcccta tcagtgatag agaacgtata aggagtttac
     8401 tccctatcag tgatagagaa cgtatgacca gtttactccc tatcagtgat agagaacgta
     8461 tctacagttt actccctatc agtgatagag aacgtatatc cagtttactc cctatcagtg
     8521 atagagaacg tataagcttt aggcgtgtac ggtgggcgcc tataaaagca gagctcgttt
     8581 agtgaaccgt cagatcgcct ggagcaattc cacaacactt ttgtcttata ccaactttcc
     8641 gtaccacttc ctaccctcgt aaaggtctag agctagccac catggagtct tctgctaaaa
     8701 tggagtccgg aggcgcggga caacaaccac aaccgcaacc acaacaaccc ttcctgccgc
     8761 cggccgcatg ttttttcgcg accgctgctg ctgctgcagc ggcggcggct gctgccgccg
     8821 cgcaatccgc ccaacagcaa caacaacaac agcagcagca gcaacaagcg cctcaacttc
     8881 gacccgctgc agacgggcag ccctcagggg gagggcacaa gagcgctccg aagcaggtta
     8941 aaaggcagag gagcagtagt cccgaactga tgcgatgtaa gaggcgcctc aattttagcg
     9001 gttttggtta ctctttgccc cagcagcagc cggctgccgt agctcgccga aatgagcggg
     9061 aaaggaaccg cgttaaactt gtgaatctcg gtttcgcgac acttcgagag cacgtaccaa
     9121 atggggcagc taacaagaaa atgagtaaag ttgagacact gcggtctgca gtggagtata
     9181 ttagagctct tcaacaattg cttgacgagc acgatgccgt atcagccgca tttcaagccg
     9241 gggtgctgtc cccaacaata tctccgaact acagcaatga tcttaatagc atggcgggaa
     9301 gtcccgtttc ctcctactcc tctgatgagg gcagctacga ccctctcagt cccgaggagc
     9361 aagagcttct tgacttcact aactggttcg gatccggcgc aacaaacttc tctctgctga
     9421 aacaagccgg agatgtcgaa gagaatcctg gaccgatgac tggagtcttt gacagtctag
     9481 tggctgatat gcactcgacc cagatcgccg cctccagcac gtaccaccag caccagcagc
     9541 ccccgagcgg cggcggcgcc ggcccgggtg gcaacagcag cagcagcagc agcctccaca
     9601 agccccagga gtcgcccacc cttccggtgt ccaccgccac cgacagcagc tactacacca
     9661 accagcagca cccggcgggc ggcggcggcg gcgggggctc gccctacgcg cacatgggtt
     9721 cctaccagta ccaagccagc ggcctcaaca acgtccctta ctccgccaag agcagctatg
     9781 acctgggcta caccgccgcc tacacctcct acgctcccta tggaaccagt tcgtccccag
     9841 ccaacaacga gcctgagaag gaggaccttg agcctgaaat tcggatagtg aacgggaagc
     9901 caaagaaagt ccggaaaccc cgcaccatct actccagttt ccagctggcg gctcttcagc
     9961 ggcgtttcca aaagactcaa tacttggcct tgccggagcg agccgagctg gcggcctctc
    10021 tgggcctcac ccagactcag gtcaaaatct
//
