LOCUS       sequence-260007-       13283 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Donor construct for introduction of hNIL factors to CLYBL safe 
            harbor site and iPSC differentiation to motor neuron.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13283)
  AUTHORS   Fernandopulle MS, Prestil R, Grunseich C, Wang C, Gan L, Ward ME
  TITLE     Transcription Factor-Mediated Differentiation of Human iPSCs into 
            Neurons.
  JOURNAL   Curr Protoc Cell Biol. 2018 Jun;79(1):e51. doi: 10.1002/cpcb.51. 
            Epub 2018 May 18.
  PUBMED    29924488
REFERENCE   2  (bases 1 to 13283)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Curr Protoc
            Cell Biol."; date: "2018-06"; volume: "79(1):e51. doi"; pages: " 
            10.1002/cpcb.51. Epub 2018 May 18"
COMMENT     Sequence Label: pUCM-CLYBL-hNIL
FEATURES             Location/Qualifiers
     source          1..13283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     266..285
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1391..1409
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1427..1445
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1463..1481
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1499..1517
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1535..1553
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1571..1589
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1607..1625
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1670..1694
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2614..2670
                     /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"
     regulatory      3461..3470
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3742..3795
                     /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"
     polyA_signal    5027..5148
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5064..5083)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5118..5137
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5197..5213)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5197..5213)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5210..5232)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5221..5237
                     /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(5245..5251,5252..5269,5270..5275))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5290..5311
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5428..5445)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5737..5756
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    5755..5810
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     5856..5875
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(5966..6712)
                     /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"
     primer_bind     complement(6744..6763)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          6771..7787
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(6825..6847)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     enhancer        8067..8446
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    8452..8676
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     8665..8682
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    8698..8731
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(9007..9026)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     9276..9294
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      9436..9445
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             9483..9751
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(9764..9975)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     polyA_signal    10011..10235
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(10240..11040)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="GPRIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(10351..10370)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     10961..10980
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(11035..11088)
                     /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"
     misc_feature    11112..11137
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    11138..11171
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             join(12029..12097,12098..12889)
                     /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(12247..12266)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(13060..13283,1..365)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
       61 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      121 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      181 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      241 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      301 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      361 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc gaattcgccc ttagagcaga
      421 gccaggaacc cctgtaggga aggggcagga gagccagggg catgagatgg tggacgagga
      481 agggggacag ggaagcctga gcgcctctcc tgggcttgcc aaggactcaa acccagaagc
      541 ccagagcagg gccttaggga agcgggacgg ctctgttgga gagtccagta ttgaattagt
      601 gatagataca cagaaaaaaa gcaaattgaa taggtgaatg gataaacaaa ctgtggaaca
      661 tccagacaat gaaatattca ccaataaaaa gaaatgagct atcaagccac attggctatg
      721 cccttcacat ttaaggagga accttaaatg catattgcca aattttaaaa gctagtctga
      781 aaatgctaca gactgtatga ttccaattat atgacatttt agaaaaggca aaactataga
      841 gatcagtggt tgccagcggc tcatggaggc aggatgaatt ggtggaacac agggatgctt
      901 tcagggtaat gaaaatatct tgtgtgatcc tgtaataatg gatacgtggc ataatgcatt
      961 tgttaaaacc tgtaaactag ccaggtatag tggctttttt gcacctatag tctcagctac
     1021 ttggtaggct gagacaggaa gatcacttga gcccaggact tcaagaccag cctgggtaat
     1081 atagtaagac cccatctcta ttttttaaag aaattaagaa agaaaagaaa cccatcaact
     1141 gtataacaca aagaatcgta atgtaaacta cagtcctgag taatgaaaca tttttaaaga
     1201 aataaataaa ttcaaggcaa ttattaactt caggaaaaac agtaaagtaa acataagtat
     1261 acaaatcaaa catggctcag ttgtgaaaat atttacaagg tcactttaag taacattgaa
     1321 tgttgatata atcccaaaat atatttatgt tggaaggatg aggagatatc tccaccccac
     1381 agtggtttac tccctatcag tgatagagaa cgtatgaaga gtttactccc tatcagtgat
     1441 agagaacgta tgcagacttt actccctatc agtgatagag aacgtataag gagtttactc
     1501 cctatcagtg atagagaacg tatgaccagt ttactcccta tcagtgatag agaacgtatc
     1561 tacagtttac tccctatcag tgatagagaa cgtatatcca gtttactccc tatcagtgat
     1621 agagaacgta taagctttag gcgtgtacgg tgggcgccta taaaagcaga gctcgtttag
     1681 tgaaccgtca gatcgcctgg agcaattcca caacactttt gtcttatacc aactttccgt
     1741 accacttcct accctcgtaa acttaaggtt aattaagccg ccaccatgtt cgtcaaatcc
     1801 gagaccttgg agttgaagga ggaagaggac gtgttagtgc tgctcggatc ggcctccccc
     1861 gccttggcgg ccctgacccc gctgtcatcc agcgccgacg aagaagagga ggaggagccg
     1921 ggcgcgtcag gcggggcgcg tcggcagcgc ggggctgagg ccgggcaggg ggcgcggggc
     1981 ggcgtggctg cgggtgcgga gggctgccgg cccgcacggc tgctgggtct ggtacacgat
     2041 tgcaaacggc gcccttcccg ggcgcgggcc gtctcccgag gcgccaagac ggccgagacg
     2101 gtgcagcgca tcaagaagac ccgtagactg aaggccaaca accgcgagcg aaaccgcatg
     2161 cacaacctca acgcggcact ggacgcgctg cgcgaggtgc tccccacgtt ccccgaggac
     2221 gccaagctca ccaagatcga gaccctgcgc ttcgcccaca actacatctg ggcactcacc
     2281 gagaccctgc gcctggcgga tcactgcggg ggcggcggcg ggggcctgcc gggggcgctc
     2341 ttctccgagg cagtgttgct gagcccggga ggcgccagcg ccgccctgag cagcagcgga
     2401 gacagcccct cgcccgcctc cacgtggagt tgcaccaaca gccccgcgcc gtcctcctcc
     2461 gtgtcctcca attccacctc cccctacagc tgcactttat cgcccgccag cccggccggg
     2521 tcagacatgg actattggca gcccccacct cccgacaagc accgctatgc acctcacctc
     2581 cccatagcca gggattgtat cttgggatcc ggagctacta acttcagcct gctgaagcag
     2641 gctggagacg tggaggagaa ccctggacct atgggagaca tgggagatcc accaaaaaaa
     2701 aaacgtctga tttccctatg tgttggttgc ggcaatcaga ttcacgatca gtatattctg
     2761 agggtttctc cggatttgga atggcatgcg gcatgtttga aatgtgcgga gtgtaatcag
     2821 tatttggacg agagctgtac atgctttgtt agggatggga aaacctactg taaaagagat
     2881 tatatcaggt tgtacgggat caaatgcgcc aagtgcagca tcggcttcag caagaacgac
     2941 ttcgtgatgc gtgcccgctc caaggtgtat cacatcgagt gtttccgctg tgtggcctgc
     3001 agccgccagc tcatccctgg ggacgaattt gcgcttcggg aggacggtct cttctgccga
     3061 gcagaccacg atgtggtgga gagggccagt ctaggcgctg gcgacccgct cagtcccctg
     3121 catccagcgc ggccactgca aatggcagcg gagcccatct ccgccaggca gccggccctg
     3181 cggccccacg tccacaagca gccggagaag accacccgcg tgcggactgt gctgaacgag
     3241 aagcagctgc acaccttgcg gacctgctac gccgcaaacc cgcggccaga tgcgctcatg
     3301 aaggagcaac tggtagagat gacgggcctc agtccccgtg tgatccgggt ctggtttcaa
     3361 aacaagcggt gcaaggacaa gaagcgaagc atcatgatga agcaactcca gcagcagcag
     3421 cccaatgaca aaactaatat ccaggggatg acaggaactc ccatggtggc tgccagtcca
     3481 gagagacacg acggtggctt acaggctaac ccagtggaag tacaaagtta ccagccacct
     3541 tggaaagtac tgagcgactt cgccttgcag agtgacatag atcagcctgc ttttcagcaa
     3601 ctggtcaatt tttcagaagg aggaccgggc tctaattcca ctggcagtga agtagcatca
     3661 atgtcctctc aacttccaga tacacctaac agcatggtag ccagtcctat tgaggcattg
     3721 atgcacagca gaggctctgg cgagggcaga ggcagcctgc tgacatgtgg cgacgtggaa
     3781 gagaaccctg gccccggtac catggaagcc agaggcgaac tgggccctgc cagagaatct
     3841 gccggcggag atctgctgct ggccctgctg gctagaaggg ccgacctgag aagagaaatc
     3901 cccctgtgcg ccggctgcga ccagcacatc ctggaccggt ttatcctgaa ggccctggac
     3961 agacactggc acagcaagtg cctgaagtgc agcgactgcc acacccccct ggccgagaga
     4021 tgctttagca gaggcgagag cgtgtactgc aaggacgact tcttcaagag attcggcacc
     4081 aagtgcgccg cctgccagct gggaattcct cctacacagg tcgtgcggag agcccaggac
     4141 ttcgtgtacc atctgcactg cttcgcctgc gtcgtgtgca agagacagct ggccaccggc
     4201 gacgagttct acctgatgga agatagccgg ctcgtgtgta aagccgacta cgagacagcc
     4261 aagcagagag aggccgaggc caccgccaaa agacccagaa ccaccatcac cgccaagcag
     4321 ctggaaaccc tgaagtccgc ctacaacacc agccccaagc ccgccagaca tgtgcgggaa
     4381 cagctgagca gcgagacagg cctggacatg agagtggtgc aagtgtggtt ccagaacaga
     4441 cgggccaaag agaagcggct gaagaaggac gccggcaggc agagatgggg ccagtacttc
     4501 cggaacatga agcggtccag aggcggcagc aagagcgaca aggacagcgt gcaggaaggc
     4561 caggacagcg acgccgaggt gtccttccca gatgagccta gcctggccga aatgggccca
     4621 gccaatggcc tgtatggcag cctgggagag ccaacccagg ctctgggcag accttctggc
     4681 gccctgggca acttctctct ggaacacggc ggactggccg gacccgagca gtatagagaa
     4741 ctgaggcccg gcagccctta cggcgtgcca ccttctccag ctgcccctca gtctctgcca
     4801 ggacctcagc ctctgctgtc cagcctggtg taccccgata caagcctggg cctggtgcca
     4861 tctggcgctc caggcggacc tcctccaatg agagtgctgg ccggcaatgg ccctagcagc
     4921 gatctgagca caggcagctc tggcggctac cccgattttc ctgccagccc tgcctcctgg
     4981 ctggacgaag tggatcacgc ccagttctga atcgatcgcc caccccaact tgtttattgc
     5041 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5101 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5161 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5221 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5281 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5341 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5401 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5461 gctgcggcga gcggtatcag ctcactcaaa ggacggccgc acatgtaagg aaaattttag
     5521 ggatgttaaa gaaaaaaata acacaaaaca aaatataaaa aaaatctaac ctcaagtcaa
     5581 ggcttttcta tggaataagg aatggacagc agggggctgt ttcatatact gatgacctct
     5641 ttatagccaa cctttgttca tggcagccag catatgggca tatgttgcca aactctaaac
     5701 caaatactca ttctgatgtt ttaaatgatt tgccctccca tatgtccttc cgagtgagag
     5761 acacaaaaaa ttccaacaca ctattgcaat gaaaataaat ttcctttatt agccagaagt
     5821 cagatgctca aggggcttca tgatgtcccc ataatttttg gcagagggaa aaagatctca
     5881 gtggtatttg tgagccaggg cattggccac accagtcacc accttctgat aggcagcctg
     5941 cacctgagga gtgaattcac gcgttttagc ctggcagcat atcgagatcg aagtcatcga
     6001 gagcgtcagc agggagcata tccagatcaa aatcatccag ggcatcggcg ggcagcatgt
     6061 ccaggtcgaa gtcgtccagg gcgtcggtgg gtccgccgct ctcgcacttc agctgcttct
     6121 cgaggccgca gatgatcagt tccaggccga acaggaaggc gggctcggcg ccctgtctgt
     6181 cgaacagctc gatggcctgc ttcagcaggg ggggcatgct gtcggtggtg ggtgtctctc
     6241 tttcctcttt ggcgacctgg tgttcctgtt cttccagcac gcagcccagg gtgaagtggc
     6301 ccacggcgct cagggcgtac agggcgtttt ccaggctgaa gccctgctgg cacaggaagg
     6361 ccagctggtt ttccagtgtc tcgtactgct tctcggtggg tctggtgccc aggtgcactt
     6421 tggcgccgtc ccggtggctc agcagggcgc atctgtagct cttggcgttg ttccgcagga
     6481 agtcctgcca gctctcgcct tccagagggc agctgtgggt gtggtgccgg tccagcatct
     6541 cgatgggcag ggcgtccagc agggcccgct tgttcttcac gtgccagtac agggtgggct
     6601 gttccacgcc cagtttctgg gccagcttcc gggtggtcag gccctcgatg cccacgccgt
     6661 tcagcagttc cagggcgctg ttgatcactt tgctcttgtc cagccggctc atgtcgacat
     6721 tctttgccaa aatgatgaga cagcacaata accagcacgt tgcccaggag ctgtaggaaa
     6781 aagaagaagg catgaacatg gttagcagag gctctagagc cgccggtcac acgccagaag
     6841 ccgaaccccg ccctgccccg tcccccccga aggcagccgt ccccccgcgg acagccccga
     6901 ggctggagag ggagaagggg acggcggcgc ggcgacgcac gaaggccctc cccgcccatt
     6961 tccttcctgc cggcgccgca ccgcttcgcc ccgcgcccgc tagagggggt gcggcggcgc
     7021 ctcccagatt tcggctccgc acagatttgg gacaaaggaa gtccctgcgc cctctcgcac
     7081 gattaccata aaaggcaatg gctgcggctc gccgcgcctc gacagccgcc ggcgctccgg
     7141 gggccgccgc gccccycccc cgagcccycc ccggcccgag gcggccccgc cccgcccggc
     7201 acccccacct gccgccaccc cccgcccggc acggcgagcc ccgcgccacg ccccgtacgg
     7261 agccccgcac ccgaagccgg gccgtgctca gcaactcggg gaggggggtg cagggggggt
     7321 tgcagcccga ccgacgcgcc cacaccccct gctcaccccc ccacgcacac accccgcacg
     7381 cagcctttgt tcccctcgca gccccccccg caccgcgggg caccgccccc ggccgcgctc
     7441 ccctcgcgca cactgcggag cgcacaaagc cccgcgccgc gcccgcagcg ctcacagccg
     7501 ccgggcagcg cggagccgca cgcggcgctc cccacgcaca cacacacgca cgcacccccc
     7561 gagccgctcc ccccgcacaa agggccctcc cggagcccct caaggctttc acgcagccac
     7621 agaaaagaaa caagccgtca ttaaaccaag cgctaattac agcccggagg agaagggccg
     7681 tcccgcccgc tcacctgtgg gagtaacgcg gtcagtcaga gccggggcgg gcggcgcgag
     7741 gcggcggcgg agcggggcac ggggcgaagg cagcgcgcag cgactcccgc ccgccgcgcg
     7801 cttcgctttt tatagggccg ccgccgccgc cgcctcgcca taaaaggaaa ctttcggagc
     7861 gcgccgctct gattggctgc cgccgcacct cyccgccycs ccccsccccc ccccccsccc
     7921 cccccccccc cccccggcgc gcsccccccc cccccccccg cccccatcgc tgcacaaaat
     7981 aattaaaaaa taaataaata caaaattggg ggtggggagg ggggggagat ggggagagtg
     8041 aagcagaacg tggggctcac ctcgaccatg gtaatagcga tgactaatac gtagatgtac
     8101 tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta
     8161 ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag
     8221 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
     8281 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
     8341 ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat
     8401 gaccccgtaa ttgattacta ttaataacta gtcaataatc aatgtcgaga cccatagagc
     8461 ccaccgcatc cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc
     8521 ccaccccacc ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat
     8581 tttattagga aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg
     8641 ggcaaacaac agatggctgg caactagaag gcacagtcga ggctgatcag cgggtttata
     8701 acttcgtata atgtatgcta tacgaagtta tggtacctta ctcgtccatg ccgccggtgg
     8761 agtggcggcc ctcggcgcgt tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg
     8821 tgatgtccaa cttgatgttg acgttgtagg cgccgggcag ctgcacgggc ttcttggcct
     8881 tgtaggtggt cttgacctca gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct
     8941 tgatctcgcc cttcagggcg ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc
     9001 ccatggtctt cttctgcatt acggggccgt cggaggggaa gttggtgccg cgcagcttca
     9061 ccttgtagat gaactcgccg tcctgcaggg aggagtcctg ggtcacggtc accacgccgc
     9121 cgtcctcgaa gttcatcacg cgctcccact tgaagccctc ggggaaggac agcttcaagt
     9181 agtcggggat gtcggcgggg tgcttcacgt aggccttgga gccgtacatg aactgagggg
     9241 acaggatgtc ccaggcgaag ggcagggggc cacccttggt caccttcagc ttggcggtct
     9301 gggtgccctc gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttca
     9361 cggagccctc catgtgcacc ttgaagcgca tgaactcctt gatgatggcc atgttatcct
     9421 cctcgccctt gctcaccatg gtggcgaccg gtagcgctag cgtaggcgcc ggtcacagct
     9481 tggatctgta acggcgcaga acagaaaacg aaacaaagac gtagagttga gcaagcaggg
     9541 tcaggcaaag cgtggagagc cggctgagtc taggtaggct ccaagggagc gccggacaaa
     9601 ggcccggtct cgacctgagc tttaaactta cctagacggc ggacgcagtt caggaggcac
     9661 cacaggcggg aggcggcaga acgcgactca accggcgtgg atggcggcct caggtagggc
     9721 ggcgggcgcg tgaaggagag atgcgagccc ctcgaagctt cagctgtgtt ctggcggcaa
     9781 acccgttgcg aaaaagaacg ttcacggcga ctactgcact tatatacggt tctcccccac
     9841 cctcgggaaa aaggcggagc cagtacacga catcactttc ccagtttacc ccgcgccacc
     9901 ttctctaggc accggttcaa ttgccgaccc ctccccccaa cttctcgggg actgtgggcg
     9961 atgtgcgctc tgcccactga cgggcaccgg agcgatcgca gatcatgcat ccatagagcc
    10021 caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc
    10081 caccccaccc cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt
    10141 ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg
    10201 gcaaacaaca gatggctggc aactagaagg cacaggtttt tagaagaact cgtcaagaag
    10261 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg
    10321 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg
    10381 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc
    10441 caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg
    10501 catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc
    10561 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg
    10621 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc
    10681 atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc
    10741 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc
    10801 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc
    10861 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag
    10921 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag
    10981 cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcctagggcc
    11041 gggattctcc tccacgtcac cgcatgttag aagacttcct ctgccctctc cgctgccaga
    11101 tctctcgagg ccctgtggga ggaagagaag aggtcagata acttcgtata atgtatgcta
    11161 tacgaagtta tgcggccgct gggagaaggg taagagagct caatcctcat cttacataga
    11221 aggaagtcag tagataatat cttacattta aaagtcaaga aataacagtc tgaccatact
    11281 atctagaaat atggaagtta acagtaagga agttaaaaca gttgaaagta gtttcgtcta
    11341 gaaagaaaga cttggggaag aacaaagaag gctaattttc attatgagct ttattgtaac
    11401 agaaaagaga ccactaactc agctaggttt tcccaattcc tcaacacttt ggggcacagt
    11461 gtttagtcag agctgtgatc actctctgag cacatggatt gtatcttaat tttcatctca
    11521 ttgaaaatat gtaccatttg tccaagtgga aactgggctg aagcatgact gtgaccacca
    11581 ccaaagaaat ggctagactc gctaaggtct gctggggtga cccgtgattg tgttgtgctt
    11641 gatcttgttg agcatggctc ttttaagtca ccatacctgc ccctgccaat gcaccaaaca
    11701 ccaacttaaa gggactttca gcttttgtaa gtcagtctca gccttacttc tgaaccattt
    11761 ctttgacttc tttcatgact ctgatcctgt cccagtttta agcctcccag ccaccagtca
    11821 tgctgttttg ctctgctatt tacattcttc tctacctact ttggacttga tggtatgcct
    11881 actatattag ttatttattg ctgtgtaaca aaccactctg aaacatagtg gcttacaacc
    11941 acaaccattt tattttatta tgaaactgtg ggtcagaatg tatattggcc cattggcatg
    12001 attctcctgc tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
    12061 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
    12121 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
    12181 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
    12241 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
    12301 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
    12361 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
    12421 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
    12481 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
    12541 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
    12601 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
    12661 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
    12721 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
    12781 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
    12841 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
    12901 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
    12961 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
    13021 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
    13081 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
    13141 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
    13201 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
    13261 tacatacctc gctctgctaa tcc
//
