LOCUS       sequence-486354-       13368 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Integrates dox-inducible hNGN2 into the human AASV1 safe harbor 
            locus for directed differentiation of neural cells from stem cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13368)
  TITLE     SNaPs 2.0 NGN2 construct
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 13368)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: 
            pGEP116[EF1a-mNeonGreen]-pAAVS1-SA-Neo-TRE3Gs-NGN2-2A-Zeo-CAG-TETON3
            G
FEATURES             Location/Qualifiers
     source          1..13368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    232..253
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(268..273,274..291,292..298)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    306..322
                     /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     311..333
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     330..346
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     330..346
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   837..871
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     misc_feature    881..1717
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        1746..2113
                     /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    1754..1772
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1790..1808
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1826..1844
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1862..1880
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1898..1916
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1934..1952
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1970..1988
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             2956..3012
                     /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             3019..3393
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3402..3626
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    3627..3660
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        3745..4048
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          4295..5312
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     5236..5258
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     5320..5339
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             5378..6127
                     /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
                     DALDDFDLDMLPADALDDFDLDMLPGS"
     polyA_signal    6136..6360
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6675..6694
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    6693..6748
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     6794..6813
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(7701..7757)
                     /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"
     misc_feature    7781..7806
                     /label=SA
                     /note="splice acceptor site"
     misc_feature    7820..8623
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     repeat_region   9087..9149
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9279..9295)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9288..9310)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9437..9892
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9524..9543)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9734..9755
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9918..10022
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10023..10091,10092..10883)
                     /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(10241..10260)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11054..11642
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11543..11562
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        11703..11914
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             11939..11959
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11963..11983
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11987..12007
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      12020..12029
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             12026..12736
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      12281..12290
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    12761..13349
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(12814..12834)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(13232..13243)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(13352..13368)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      181 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      241 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      301 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
      361 gcgcgcccgc cgggtaactc acggggtatc catgtccatt tctgcggcat ccagccagga
      421 tacccgtcct cgctgacgta atatcccagc gccgcaccgc tgtcattaat ctgcacaccg
      481 gcacggcagt tccggctgtc gccggtattg ttcgggttgc tgatgcgctt cgggctgacc
      541 atccggaact gtgtccggaa aagccgcgac gaactggtat cccaggtggc ctgaacgaac
      601 agttcaccgt taaaggcgtg catggccaca ccttcccgaa tcatcatggt aaacgtgcgt
      661 tttcgctcaa cgtcaatgca gcagcagtca tcctcggcaa actctttcca tgccgcttca
      721 acctcgcggg aaaaggcacg ggcttcttcc tccccgatgc ccagatagcg ccagcttggg
      781 cgatgactga gccggaaaaa agacccgacg atatgatcct gatgcagcta gattaaccct
      841 agaaagatag tctgcgtaaa attgacgcat gcatctcgag agagcagagc caggaacacc
      901 tgtagggaag gggcaggaga gccaggggca tgagatggtg gacgaggaag ggggacaggg
      961 aagcctgagc gcctctcctg ggcttgccaa ggactcaaac ccagaagccc agagcagggc
     1021 cttagggaag cgggaccctg ctctgggcgg aggaatatgt cccagatagc actggggact
     1081 ctttaaggaa agaaggatgg agaaagagaa agggagtaga ggcggccacg acctggtgaa
     1141 cacctaggac gcaccattct cacaaaggga gttttccaca cggacacccc cctcctcacc
     1201 acagccctgc caggacgggg ctggctactg gccttatctc acaggtaaaa ctgacgcacg
     1261 gaggaacaat ataaattggg gactagaaag gtgaagagcc aaagttagaa ctcaggacca
     1321 acttattctg attttgtttt tccaaactgc ttctcctctt gggaagtgta aggaagctgc
     1381 agcaccagga tcagtgaaac gcaccagaca gccgcgtcag agcagctcag gttctgggag
     1441 agggtagcgc agggtggcca ctgagaaccg ggcaggtcac gcatcccccc cttccctccc
     1501 accccctgcc aagctctccc tcccaggatc ctctctggct ccatcgtaag caaaccttag
     1561 aggttctggc aaggagagag atggctccag gaaatggggg tgtgtcacca gataaggaat
     1621 ctgcctaaca ggaggtgggg gttagaccca atatcaggag actaggaagg aggaggccta
     1681 aggatggggc ttttctgtca ccaatcctgt ccctagtagt gtacatgcca gaacatttct
     1741 ccctcgagtt tactccctat cagtgataga gaacgtatga agagtttact ccctatcagt
     1801 gatagagaac gtatgcagac tttactccct atcagtgata gagaacgtat aaggagttta
     1861 ctccctatca gtgatagaga acgtatgacc agtttactcc ctatcagtga tagagaacgt
     1921 atctacagtt tactccctat cagtgataga gaacgtatat ccagtttact ccctatcagt
     1981 gatagagaac gtataagctt tgcttatgta aaccagggcg cctataaaag agtgctgatt
     2041 ttttgagtaa acttcaattc cacaacactt ttgtcttata ccaactttcc gtaccacttc
     2101 ctaccctcgt aaagccgggc gcgccccacc atgttcgtca agagcgaaac cctggagctg
     2161 aaggaggagg aggacgtgct ggtgctgctg ggatctgcat cacctgccct ggctgctctg
     2221 accccactga gctcctctgc cgacgaagaa gaggaggagg agcccggcgc ctccggcggc
     2281 gcccggagac agcgcggcgc cgaggccggc cagggagcaa ggggaggagt ggcagcagga
     2341 gccgagggct gccgccctgc ccggctgctg ggcctggtgc acgattgtaa gaggcgccca
     2401 tctagggcaa gggccgtgag caggggagca aagaccgccg agacagtgca gcggatcaag
     2461 aagacacgga gactgaaggc caacaatcgg gagagaaaca ggatgcacaa cctgaatgcc
     2521 gccctggacg cactgaggga ggtgctgcct accttcccag aggatgccaa gctgaccaag
     2581 atcgagacac tgagatttgc ccacaattac atctgggcac tgaccgagac actgaggctg
     2641 gcagaccact gcggaggagg aggaggagga ctgccaggcg ccctgttctc cgaggccgtg
     2701 ctgctgagcc ctggaggagc atccgccgcc ctgagctcct ctggcgattc tcctagccca
     2761 gcctctacct ggagctgcac aaactctccc gcccctagct cctctgtgag ctccaattcc
     2821 acatctccct attcttgtac cctgtcccca gcatctcctg caggaagcga catggattac
     2881 tggcagcccc ctccacccga caagcacaga tatgccccac acctgcccat cgccagggat
     2941 tgtatcggct ccggcgccac aaacttttct ctgctgaagc aggcaggcga cgtggaggag
     3001 aatccaggac ccgtcgacat ggccaagttg accagtgccg ttccggtgct caccgcgcgc
     3061 gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg
     3121 gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag
     3181 gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg
     3241 tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg
     3301 accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac
     3361 tgcgtgcact tcgtggccga ggagcaggac taattaatta actgtgcctt ctagttgcca
     3421 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3481 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3541 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3601 tgctggggat gcggtgggct ctatggataa cttcgtataa tgtatgctat acgaagttat
     3661 gcccgggcaa ggatctgcga tcgctagtta ttaatagtaa tcaattacgg ggtcattagt
     3721 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     3781 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     3841 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     3901 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     3961 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     4021 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
     4081 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
     4141 tgcagcgatg ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc
     4201 ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg
     4261 gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc
     4321 cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact
     4381 cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta
     4441 atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagggccc
     4501 tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc
     4561 gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc
     4621 gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt gcgggggggg
     4681 ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt
     4741 gggcgcgtcg gtcgggctgc aaccccccct gcacccccct ccccgagttg ctgagcacgg
     4801 cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg
     4861 ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg gggagggctc
     4921 gggggagggg cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag
     4981 ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg
     5041 tgcggagccg aaatctggga ggcgccgccg caccccctct agcgggcgcg gggcgaagcg
     5101 gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg
     5161 tccccttctc cctctccagc ctcggggctg tccgcggggg gacggctgcc ttcggggggg
     5221 gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta
     5281 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc
     5341 tgtctcatca ttttggcaaa gaattgcggc cgccaccatg tctagactgg acaagagcaa
     5401 agtcataaac tctgctctgg aattactcaa tggagtcggt atcgaaggcc tgacgacaag
     5461 gaaactcgct caaaagctgg gagttgagca gcctaccctg tactggcacg tgaagaacaa
     5521 gcgggccctg ctcgatgccc tgccaatcga gatgctggac aggcatcata cccactcctg
     5581 ccccctggaa ggcgagtcat ggcaagactt tctgcggaac aacgccaagt cataccgctg
     5641 tgctctcctc tcacatcgcg acggggctaa agtgcatctc ggcacccgcc caacagagaa
     5701 acagtacgaa accctggaaa atcagctcgc gttcctgtgt cagcaaggct tctccctgga
     5761 gaacgcactg tacgctctgt ccgccgtggg ccactttaca ctgggctgcg tattggagga
     5821 acaggagcat caagtagcaa aagaggaaag agagacacct accaccgatt ctatgccccc
     5881 acttctgaaa caagcaattg agctgttcga ccggcaggga gccgaacctg ccttcctttt
     5941 cggcctggaa ctaatcatat gtggcctgga gaaacagcta aagtgcgaaa gcggcgggcc
     6001 gaccgacgcc cttgacgatt ttgacttaga catgctccca gccgatgccc ttgacgactt
     6061 tgaccttgat atgctgcctg ctgacgctct tgacgatttt gaccttgaca tgctccccgg
     6121 gtcataggtt taaacctgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     6181 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     6241 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     6301 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     6361 cttaaggatc tccataagag aagagggaca gctatgactg ggagtagtca ggagaggagg
     6421 aaaaatctgg ctagtaaaac aggtaaggaa aattttaggg atgttaaaga aaaaaataac
     6481 acaaaacaaa atataaaaaa aatctaacct caagtcaagg cttttctatg gaataaggaa
     6541 tggacagcag ggggctgttt catatactga tgacctcttt atagccaacc tttgttcatg
     6601 gcagccagca tatgggcata tgttgccaaa ctctaaacca aatactcatt ctgatgtttt
     6661 aaatgatttg ccctcccata tgtccttccg agtgagagac acaaaaaatt ccaacacact
     6721 attgcaatga aaataaattt cctttattag ccagaagtca gatgctcaag gggcttcatg
     6781 atgtccccat aatttttggc agagggaaaa agatcacagt ggtatttgtg agccagggca
     6841 ttggccacac cagccaccac cttctgatag gcagcctgca cctgaggagt gttaacctat
     6901 taaaagaact catccagcag tctgtaaaaa gcaatcctct gtgaatcagg tgcggcaatc
     6961 ccatacagca ccaggaacct atcggcccat tccccgccca gttcctctgc aatgtctctt
     7021 gtggccaggg cgatgtcctg gtatctgtcg gcgacgccca gtcgtccaca atcaataaac
     7081 ccactgaatc tgccattctc caccatgatg ttgggcaggc aagcgtcgcc atgggtgacc
     7141 accagatctt ctccgtctgg cattctggct ttcagccgtg caaacagctc tgcaggagcc
     7201 agcccctggt gttcctcatc cagatcgtcc tggtccacca gcccggcctc cattcgggtt
     7261 cgagccctct caatgcgatg tttagcctgg tgatcgaaag ggcaggtggc ggggtccaga
     7321 gtatgcagtc tccgcatggc atctgccatg atggagacct tctcagctgg tgccaggtgg
     7381 gagctcagca ggtcctgtcc tggcacttct cccagcagca gccaatctcg cccggcctca
     7441 gtgaccacgt ccaggactgc agcacaaggc actcctgtgg tggccagcca gctcaggcgg
     7501 gctgcttcat cctgcagctc gttcagtgct cctgacaggt ctgtcttgac aaacagcacg
     7561 gggcgcccct gagcactcag ccggaacacg gctgcgtcag agcatccaat agtctgctgt
     7621 gcccaatcgt atccaaacag tctttcgacc caagcggctg gtgacccggc gtgcagtccg
     7681 tcctgctcaa tcatgatatc aggaccgggg ttttcttcca cgtctcctgc ttgctttaac
     7741 agagagaagt tcgtggcacc agagccagat ctctcgaggc cctgtgggag gaagagaaga
     7801 ggtcagaagc ttgacgcgtg ccccactgtg gggtggaggg gacagataaa agtacccaga
     7861 accagagcca cattaaccgg ccctgggaat ataaggtggt cccagctcgg ggacacagga
     7921 tccctggagg cagcaaacat gctgtcctga agtggacata ggggcccggg ttggaggaag
     7981 aagactagct gagctctcgg acccctggaa gatgccatga cagggggctg gaagagctag
     8041 cacagactag agaggtaagg ggggtagggg agctgcccaa atgaaaggag tgagaggtga
     8101 cccgaatcca caggagaacg gggtgtccag gcaaagaaag caagaggatg gagaggtggc
     8161 taaagccagg gagacggggt actttggggt tgtccagaaa aacggtgatg atgcaggcct
     8221 acaagaaggg gaggcgggac gcaagggaga catccgtcgg agaaggccat cctaagaaac
     8281 gagagatggc acaggcccca gaaggagaag gaaaagggaa cccagcgagt gaagacggca
     8341 tggggttggg tgagggagga gagatgcccg gagaggaccc agacacgggg aggatccgct
     8401 cagaggacat cacgtggtgc agcgccgaga aggaagtgct ccggaaagag catccttggg
     8461 cagcaacaca gcagagagca aggggaagag ggagtggagg aagacggaac ctgaaggagg
     8521 cggcagggaa ggatctgggc cagccgtaga ggtgacccag gccacaagct gcagacagaa
     8581 agcggcacag gcccagggga gagaatgctg gtcagagaaa gcaggtacca agggcgaatt
     8641 ccagcacact ggcggccgtt actagtggcc gtggaggcct tttccccgta tccccccagg
     8701 tgtctgcagg ctcaaagagc agcgagaagc gttcagagga aagcgatccc gtgccacctt
     8761 ccccgtgccc gggctgtccc cgcacgctgc cggctcgggg atgcgggggg agcgccggac
     8821 cggagcggag ccccgggcgg ctcgctgctg ccccctagcg ggggagggac gtaattacat
     8881 ccctgggggc tttggggggg ggctgtccct cataaaagtt ttgttacttt atagaagaaa
     8941 ttttgagttt ttgttttttt taataaataa ataaacataa ataaattgtt tgttgaattt
     9001 attattagta tgtaagtgta aatataataa aacttaatat ctattcaaat taataaataa
     9061 acctcgatat acagaccgat aaaacacatg cgtcaatttt acacatgatt atctttaacg
     9121 tacgtcacaa tatgattatc tttctagggt taatctagct gcgtgttctg cagcgtgtcg
     9181 agcatcttca tctgctccat cacgctgtaa aacacatttg caccgcgagt ctgcccgtcc
     9241 tccacgggtt caaaaacgtg aatgaacgag gcgcgctcac tggccgtcgt tttacaacgt
     9301 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     9361 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     9421 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     9481 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     9541 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     9601 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     9661 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     9721 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     9781 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     9841 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     9901 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     9961 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
    10021 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
    10081 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
    10141 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
    10201 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
    10261 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
    10321 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
    10381 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
    10441 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
    10501 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
    10561 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
    10621 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
    10681 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
    10741 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
    10801 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
    10861 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
    10921 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
    10981 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
    11041 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
    11101 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
    11161 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
    11221 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
    11281 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
    11341 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
    11401 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
    11461 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
    11521 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
    11581 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
    11641 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
    11701 tggggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
    11761 atccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
    11821 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
    11881 tctttttcgc aacgggtttg ccgccagaac acaggaccgg tccaccatgg gatcagatcc
    11941 aaaaaagaag agaaaggtag acccgaagaa aaagcgtaag gtcgatccca aaaagaaacg
    12001 gaaagtggga tcaccagtcg ccaccatggt gagcaagggc gaggaggata acatggcctc
    12061 tctcccagcg acacatgagt tacacatctt tggctccatc aacggtgtgg actttgacat
    12121 ggtgggtcag ggcaccggca atccaaatga tggttatgag gagttaaacc tgaagtccac
    12181 caagggtgac ctccagttct ccccctggat tctggtccct catatcgggt atggcttcca
    12241 tcagtacctg ccctaccctg acgggatgtc gcctttccag gccgccatgg tagatggctc
    12301 cggctaccaa gtccatcgca caatgcagtt tgaagatggt gcctccctta ctgttaacta
    12361 ccgctacacc tacgagggaa gccacatcaa aggagaggcc caggtgaagg ggactggttt
    12421 ccctgctgac ggtcctgtga tgaccaactc gctgaccgct gcggactggt gcaggtcgaa
    12481 gaagacttac cccaacgaca aaaccatcat cagtaccttt aagtggagtt acaccactgg
    12541 aaatggcaag cgctaccgga gcactgcgcg gaccacctac acctttgcca agccaatggc
    12601 ggctaactat ctgaagaacc agccgatgta cgtgttccgt aagacggagc tcaagcactc
    12661 caagaccgag ctcaacttca aggagtggca aaaggccttt accgatgtga tgggcatgga
    12721 cgagctgtac aagtaagaat tcgatatcaa gcttatcggt aatcaacctc tggattacaa
    12781 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
    12841 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
    12901 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
    12961 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
    13021 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
    13081 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
    13141 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
    13201 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
    13261 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
    13321 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcg
//
