LOCUS       sequence-388955-       15442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac plasmid with tet-inducible expression of transcription 
            factors for sensory neuron differentiation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15442)
  AUTHORS   Held A, Adler M, Marques C, Reyes CJ, Kavuturu AS, Quadros ARAA, 
            Ndayambaje IS, Lara E, Ward M, Lagier-Tourenne C, Wainger BJ
  TITLE     iPSC motor neurons, but not other derived cell types, capture gene 
            expression changes in postmortem sporadic ALS motor neurons.
  JOURNAL   Cell Rep. 2023 Aug 30;42(9):113046. doi: 
            10.1016/j.celrep.2023.113046.
  PUBMED    37651231
REFERENCE   2  (bases 1 to 15442)
  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: "Cell Rep.";
            date: "2023-08-30"; volume: "42(9):113046. doi"; pages: " 
            10.1016/j.celrep.2023.113046"
COMMENT     Sequence Label: PB-tetO-NB
FEATURES             Location/Qualifiers
     source          1..15442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(441..1232,1233..1301))
                     /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     1064..1083
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1302..1406)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1432..1887)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1569..1590)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1781..1800
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2014..2036
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2028..2045
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2029..2045
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   2175..2237
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        3342..3719
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    3349..3367
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3385..3403
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3421..3439
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3457..3475
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3493..3511
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3529..3547
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3565..3583
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3628..3652
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4572..4625
                     /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             4929..4955
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     polyA_signal    5939..6060
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5976..5995)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6030..6049
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6109..6125)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6133..6149
                     /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(6157..6163,6164..6181,6182..6187))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6202..6223
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     6649..6668
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    6667..6722
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     6768..6787
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(6878..7624)
                     /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(7656..7675)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          7683..8699
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(7737..7759)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(8700..8977)
                     /label=chicken beta-actin promoter
     enhancer        8979..9358
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    9359..9407
                     /note="synthetic polyadenylation signal"
     misc_feature    9421..9512
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     9461..9480
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     protein_bind    complement(10724..10757)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        10758..10969
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          10970..11908
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     11865..11885
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             11931..12527
                     /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(11931..11950)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     12427..12447
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             12537..12590
                     /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             12597..12623
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             12639..12665
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     regulatory      12687..12696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             12693..13403
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     polyA_signal    13423..13647
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(13648..13681)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   14197..14231
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(14722..14738)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    14746..14762
                     /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(14770..14776,14777..14794,14795..14800))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14815..14836
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(join(15124..15442,1..270))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(15204..15223)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
       61 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      121 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      181 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      241 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      301 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      361 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      421 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
      481 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
      541 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
      601 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
      661 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
      721 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
      781 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
      841 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
      901 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
      961 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1021 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1081 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1141 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1201 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1261 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1321 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1381 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     1441 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     1501 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     1561 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     1621 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     1681 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     1741 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     1801 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     1861 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     1921 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     1981 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     2041 ccagtgagcg cgcctcgttc attcacgttt ttgaacccgt ggaggacggg cagactcgcg
     2101 gtgcaaatgt gttttacagc gtgatggagc agatgaagat gctcgacacg ctgcagaaca
     2161 cgcagctaga ttaaccctag aaagataatc atattgtgac gtacgttaaa gataatcatg
     2221 tgtaaaattg acgcatgtgt tttatcggtc tgtatatcga ggtttattta ttaatttgaa
     2281 tagatattaa gttttattat atttacactt acatactaat aataaattca acaaacaatt
     2341 tatttatgtt tatttattta ttaaaaaaaa caaaaactca aaatttcttc tataaagtaa
     2401 caaaactttt atgagggaca gccccccccc aaagccccca gggatgtaat tacgtccctc
     2461 ccccgctagg gggcagcagc gagccgcccg gggctccgct ccggtccggc gctccccccg
     2521 catccccgag ccggcagcgt gcggggacag cccgggcacg gggaaggtgg cacgggatcg
     2581 ctttcctctg aacgcttctc gctgctcttt gagcctgcag acacctgggg ggatacgggg
     2641 aaaaggcctc cacggcctta attaagggag gtggtccctg cagttacgcc aatgataacc
     2701 cccgccagaa aaatcttagt agccttccct ttttgttttc cgtgccccaa ctcggcggat
     2761 tgactcggcc ccttccggaa acacccgaat caacttctag tcaaattatt gttcacgccg
     2821 caatgaccca cccctggccc gcgtctgtgg aactgacccc tggtgtacag gagagttcgc
     2881 tgctgaaagt ggtcccaaag gggtactagt ttttaagctc ccaactcccc ctcccccagc
     2941 gtctggagga ttccacaccc tcgcaccgca ggggcgagga agtgggcgga gtccggtttt
     3001 ggcgccagcc gctgaggctg ccaagcagaa aagccaccgc tgaggagact ccggtcactg
     3061 tcctcgcccc gcctccccct tccctcccct tggggaccac cgggcgccac gccgcgaacg
     3121 gtaagtgccg cggtcgtcgg cgcctccgcc ctccccctag ggccccaatt cccagcgggc
     3181 gcggcgcgcg gcccctcccc ccgccgggcg cgcgcccgct gccccgccct tcgtggccgc
     3241 ccggcgtggg cggtgccacc cctccccccg gcggccccgc gcgcagctcc cggctccctc
     3301 ccccttcgga tgtggcttga gctgtaggcg cggagggtac cagtttactc cctatcagtg
     3361 atagagaacg tatgaagagt ttactcccta tcagtgatag agaacgtatg cagactttac
     3421 tccctatcag tgatagagaa cgtataagga gtttactccc tatcagtgat agagaacgta
     3481 tgaccagttt actccctatc agtgatagag aacgtatcta cagtttactc cctatcagtg
     3541 atagagaacg tatatccagt ttactcccta tcagtgatag agaacgtata agctttaggc
     3601 gtgtacggtg ggcgcctata aaagcagagc tcgtttagtg aaccgtcaga tcgcctggag
     3661 caattccaca acacttttgt cttataccaa ctttccgtac cacttcctac cctcgtaaag
     3721 gatccgccgc caccatgttc gtgaagtccg agacactgga actgaaagag gaagaggacg
     3781 tgctggtgct gctgggctct gcctctcctg ctctggctgc tctgacaccc ctgagcagca
     3841 gcgccgacga ggaagaagaa gaggaacctg gcgcaagcgg cggagccaga aggcagagag
     3901 gcgctgaagc tggacagggc gcaagaggcg gagtggctgc tggcgcagaa ggatgcagac
     3961 ctgccagact gctgggcctg gtgcacgact gcaagagaag gccctctaga gccagagccg
     4021 tgtccagagg cgccaagaca gccgagacag tgcagcggat caagaaaacc agacggctga
     4081 aggccaacaa ccgcgagcgg aaccggatgc acaacctgaa cgccgccctg gacgccctga
     4141 gagaggtgct gcctaccttc cccgaggacg ccaagctgac caagatcgaa accctgagat
     4201 tcgcccacaa ctacatctgg gccctgaccg agactctgcg gctggccgat cattgtggcg
     4261 gcggaggcgg aggactgcca ggcgctctgt tttctgaagc cgtgctgctg tctcctggcg
     4321 gagcttctgc tgccctgtct agcagcggcg attctcctag ccctgccagc acctggtcct
     4381 gcaccaatag cccagcccct agctccagcg tgtccagcaa ttccaccagc ccttacagct
     4441 gcaccctgag ccctgcttct ccagccggca gcgacatgga ctactggcag cctccccccc
     4501 ctgacaaaca tcgctacgcc ccccatctgc ctatcgcccg ggattgcatc atgcacagca
     4561 gaggctctgg cgagggcaga ggcagcctgc tgacatgtgg cgacgtggaa gagaaccctg
     4621 gccccggttc catgatgtcc atgaactcca aacagccaca ctttgccatg cacccaactc
     4681 tgcctgagca taagtaccca tccctgcatt cctcctccga ggcaatcagg agggcttgtc
     4741 tgcctactcc tcctctccag agcaacctgt tcgcttctct ggacgaaaca ctgctcgcaa
     4801 gagccgaagc cctcgcagcc gtggacattg ctgtttctca gggcaagtcc catcctttca
     4861 aacccgacgc cacctaccac accatgaata gcgtcccatg caccagcacc tccactgtcc
     4921 cactcgctca tcatcatcac catcaccacc accaccaggc actggagcct ggtgatctcc
     4981 tcgaccacat ctcctctcca tccctggctc tgatggctgg agctggaggt gctggagcag
     5041 ctgcaggtgg aggtggagcc cacgacggtc caggaggagg tggaggacca ggaggaggag
     5101 gaggacctgg aggtggtcct ggtggtggag gaggtggcgg gccaggagga ggtggtggcg
     5161 ggccaggtgg aggactcctc ggaggaagcg ctcaccctca tcctcacatg cacagcctgg
     5221 gtcacctgtc ccaccctgct gctgctgctg ctatgaacat gccttctgga ctgcctcacc
     5281 ctggcctggt tgcagcagct gctcatcacg gtgcagctgc tgcagctgca gctgctgctg
     5341 caggtcaggt cgctgctgcc tctgctgcag ctgccgtcgt tggagctgct ggactggcct
     5401 ccatctgcga ttccgacaca gacccaagag aactcgaagc attcgctgaa agattcaaac
     5461 aaagaagaat caaactcggt gttactcaag cagatgtggg aagcgcactg gctaacctca
     5521 agatccctgg agtcggcagc ctgagccagt ccaccatctg tcgctttgaa tctctgaccc
     5581 tgtcccacaa caacatgatt gccctcaagc ctatcctcca ggcttggctg gaggaggcag
     5641 agggagcaca acgcgagaag atgaacaagc cagagctgtt caacggaggc gagaagaagc
     5701 gcaagcgcac ttccattgct gcacctgaga agagatccct ggaggcatac ttcgcagtgc
     5761 agcccaggcc atcttctgag aagatcgcag ctattgccga gaagctcgac ctcaagaaga
     5821 acgtggtcag ggtgtggttc tgcaatcaga ggcagaaaca gaagagaatg aagttctccg
     5881 caacctactg agctagctga actcgagtgt ttaaactcgc gaatcgatcg cccaccccaa
     5941 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6001 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     6061 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     6121 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6181 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6241 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6301 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6361 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggacggcc gcacatgtaa
     6421 ggaaaatttt agggatgtta aagaaaaaaa taacacaaaa caaaatataa aaaaaatcta
     6481 acctcaagtc aaggcttttc tatggaataa ggaatggaca gcagggggct gtttcatata
     6541 ctgatgacct ctttatagcc aacctttgtt catggcagcc agcatatggg catatgttgc
     6601 caaactctaa accaaatact cattctgatg ttttaaatga tttgccctcc catatgtcct
     6661 tccgagtgag agacacaaaa aattccaaca cactattgca atgaaaataa atttccttta
     6721 ttagccagaa gtcagatgct caaggggctt catgatgtcc ccataatttt tggcagaggg
     6781 aaaaagatct cagtggtatt tgtgagccag ggcattggcc acaccagtca ccaccttctg
     6841 ataggcagcc tgcacctgag gagtgaattc acgcgtttta gcctggcagc atatcgagat
     6901 cgaagtcatc gagagcgtca gcagggagca tatccagatc aaaatcatcc agggcatcgg
     6961 cgggcagcat gtccaggtcg aagtcgtcca gggcgtcggt gggtccgccg ctctcgcact
     7021 tcagctgctt ctcgaggccg cagatgatca gttccaggcc gaacaggaag gcgggctcgg
     7081 cgccctgtct gtcgaacagc tcgatggcct gcttcagcag ggggggcatg ctgtcggtgg
     7141 tgggtgtctc tctttcctct ttggcgacct ggtgttcctg ttcttccagc acgcagccca
     7201 gggtgaagtg gcccacggcg ctcagggcgt acagggcgtt ttccaggctg aagccctgct
     7261 ggcacaggaa ggccagctgg ttttccagtg tctcgtactg cttctcggtg ggtctggtgc
     7321 ccaggtgcac tttggcgccg tcccggtggc tcagcagggc gcatctgtag ctcttggcgt
     7381 tgttccgcag gaagtcctgc cagctctcgc cttccagagg gcagctgtgg gtgtggtgcc
     7441 ggtccagcat ctcgatgggc agggcgtcca gcagggcccg cttgttcttc acgtgccagt
     7501 acagggtggg ctgttccacg cccagtttct gggccagctt ccgggtggtc aggccctcga
     7561 tgcccacgcc gttcagcagt tccagggcgc tgttgatcac tttgctcttg tccagccggc
     7621 tcatgtcgac attctttgcc aaaatgatga gacagcacaa taaccagcac gttgcccagg
     7681 agctgtagga aaaagaagaa ggcatgaaca tggttagcag aggctctaga gccgccggtc
     7741 acacgccaga agccgaaccc cgccctgccc cgtccccccc gaaggcagcc gtccccccgc
     7801 ggacagcccc gaggctggag agggagaagg ggacggcggc gcggcgacgc acgaaggccc
     7861 tccccgccca tttccttcct gccggcgccg caccgcttcg ccccgcgccc gctagagggg
     7921 gtgcggcggc gcctcccaga tttcggctcc gcacagattt gggacaaagg aagtccctgc
     7981 gccctctcgc acgattacca taaaaggcaa tggctgcggc tcgccgcgcc tcgacagccg
     8041 ccggcgctcc gggggccgcc gcgcccctcc cccgagccct ccccggcccg aggcggcccc
     8101 gccccgcccg gcacccccac ctgccgccac cccccgcccg gcacggcgag ccccgcgcca
     8161 cgccccgtac ggagccccgc acccgaagcc gggccgtgct cagcaactcg gggagggggg
     8221 tgcagggggg gttgcagccc gaccgacgcg cccacacccc ctgctcaccc ccccacgcac
     8281 acaccccgca cgcagccttt gttcccctcg cagccccccc cgcaccgcgg ggcaccgccc
     8341 ccggccgcgc tcccctcgcg cacactgcgg agcgcacaaa gccccgcgcc gcgcccgcag
     8401 cgctcacagc cgccgggcag cgcggagccg cacgcggcgc tccccacgca cacacacacg
     8461 cacgcacccc ccgagccgct ccccccgcac aaagggccct cccggagccc ctcaaggctt
     8521 tcacgcagcc acagaaaaga aacaagccgt cattaaacca agcgctaatt acagcccgga
     8581 ggagaagggc cgtcccgccc gctcacctgt gggagtaacg cggtcagtca gagccggggc
     8641 gggcggcgcg aggcggcggc ggagcggggc acggggcgaa ggcagcgcgc agcgactccc
     8701 gcccgccgcg cgcttcgctt tttatagggc cgccgccgcc gccgcctcgc cataaaagga
     8761 aactttcgga gcgcgccgct ctgattggct gccgccgcmc ctctccgccy csccccsccc
     8821 csccccycsc cccsccccsc cccsccyggc gcgcgccccc cccccccccc cgcccccatc
     8881 gctgcacaaa ataattaaaa aataaataaa tacaaaattg ggggtgggga ggggggggag
     8941 atggggagag tgaagcagaa cgtggggctc acctcgacca tggtaatagc gatgactaat
     9001 acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag
     9061 gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga
     9121 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
     9181 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
     9241 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
     9301 ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcaa
     9361 taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact
     9421 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
     9481 cagtgcaagt gcaggtgcca gaacatttct ctgagctcac ggggacagcc cccccccaaa
     9541 gcccccaggg atgtaattac gtccctcccc cgctaggggg cagcagcgag ccgcccgggg
     9601 ctccgctccg gtccggcgct ccccccgcat ccccgagccg gcagcgtgcg gggacagccc
     9661 gggcacgggg aaggtggcac gggatcgctt tcctctgaac gcttctcgct gctctttgag
     9721 cctgcagaca cctgggggga tacggggaaa aagctttagg ctgaaagaga gatttagaat
     9781 gacagaatca tagaacggcc tgggttgcaa aggagcacag tgctcatcca gatccaaccc
     9841 cctgctatgt gcagggtcat caaccagcag cccaggctgc ccagagccac atccagcctg
     9901 gccttgaatg cctgcaggga tggggcatcc acagcctcct tgggcaacct gttcagtgcg
     9961 tcaccaccct ctgggggaaa aactgcctcc tcatatccaa cccaaacctc ccctgtctca
    10021 gtgtaaagcc attccccctt gtcctatcaa gggggagttt gctgtgacat tgttggtctg
    10081 gggtgacaca tgtttgccaa ttcagtgcat cacggagagg cagatcttgg ggataaggaa
    10141 gtgcaggaca gcatggacgt gggacatgca ggtgttgagg gctctgggac actctccaag
    10201 tcacagcgtt cagaacagcc ttaaggataa gaagatagga tagaaggaca aagagcaagt
    10261 taaaacccag catggagagg agcacaaaaa ggccacagac actgctggtc cctgtgtctg
    10321 agcctgcatg tttgatggtg tctggatgca agcagaaggg gtggaagagc ttgcctggag
    10381 agatacagct gggtcagtag gactgggaca ggcagctgga gaattgccat gtagatgttc
    10441 atacaatcgt caaatcatga aggctggaaa agccctccaa gatccccaag accaacccca
    10501 acccacccac cgtgcccact ggccatgtcc ctcagtgcca catccccaca gttcttcatc
    10561 acctccaggg acggtgaccc ccccacctcc gtgggcagct gtgccactgc agcaccgctc
    10621 tttggagaag gtaaatcttg ctaaatccag cccgaccctc ccctggcaca acgtaaggcc
    10681 attatctctc atccaactcc aggacggagt cagtgagaat attataactt cgtatagcat
    10741 acattatacg aagttatggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
    10801 aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
    10861 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
    10921 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt
    10981 gtgtggttcc cgcgggcctg gcctctttac gggttatggc ccttgcgtgc cttgaattac
    11041 ttccacctgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa gtgggtggga
    11101 gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc
    11161 tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt
    11221 cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt ttttctggca
    11281 agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt ttggggccgc
    11341 gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc
    11401 gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg
    11461 cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt
    11521 tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa aatggaggac
    11581 gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg cctttccgtc
    11641 ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta
    11701 gttctcgcgc ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga
    11761 gtttccccac actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt
    11821 ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt
    11881 ggttcaaagt ttttttcttc catttcaggt gtcgtgattt aaatcccacc atgaccgagt
    11941 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
    12001 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
    12061 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
    12121 tgtgggtcgc ggacgacggc gccgccgtgg cggtctggac cacgccggag agcgtcgaag
    12181 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
    12241 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
    12301 tcctggccac cgtcggagtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
    12361 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
    12421 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
    12481 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgccgga tcgggagagg
    12541 gcagaggaag tctgctaaca tgcggtgacg tcgaggagaa tcctggccca ccggtccctg
    12601 ccgctaaacg ggtcaaactc gacggcggtt ctggaggtcc tgctgcaaag cgcgttaagt
    12661 tggatggagg gtcaggggga gccaccgcca ccatggtgtc taagggcgaa gagctgatta
    12721 aggagaacat gcacatgaag ctgtacatgg agggcaccgt ggacaaccat cacttcaagt
    12781 gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga atcaaggtgg
    12841 tcgagggcgg ccctctcccc ttcgccttcg acatcctggc tactagcttc ctctacggca
    12901 gcaagacctt catcaaccac acccagggca tccccgactt cttcaagcag tccttccctg
    12961 agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc
    13021 aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga ggggtgaact
    13081 tcacatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc ttcaccgaga
    13141 cgctgtaccc cgctgacggc ggcctggaag gcagaaacga catggccctg aagctcgtgg
    13201 gcgggagcca tctgatcgca aacgccaaga ccacatatag atccaagaaa cccgctaaga
    13261 acctcaagat gcctggcgtc tactatgtgg actacagact ggaaagaatc aaggaggcca
    13321 acaacgagac ctacgtcgag cagcacgagg tggcagtggc cagatactgc gacctcccta
    13381 gcaaactggg gcacaagctt aattaggcgg ccgctaagaa ttctgtgcct tctagttgcc
    13441 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
    13501 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
    13561 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
    13621 atgctgggga tgcggtgggc tctatggata acttcgtata gcatacatta tacgaagtta
    13681 tttggtatgg ctttttcccc gtatcccccc aggtgtctgc aggctcaaag agcagcgaga
    13741 agcgttcaga ggaaagcgat cccgtgccac cttccccgtg cccgggctgt ccccgcacgc
    13801 tgccggctcg gggatgcggg gggagcgccg gaccggagcg gagccccggg cggctcgctg
    13861 ctgcccccta gcgggggagg gacgtaatta catccctggg ggctttgggg gggggctgtc
    13921 cctgattcta taacaagaaa atatatatat aataagttat cacgtaagta gaacatgaaa
    13981 taacaatata attatcgtat gagttaaatc ttaaaagtca cgtaaaagat aatcatgcgt
    14041 cattttgact cacgcggtcg ttatagttca aaatcagtga cacttaccgc attgacaagc
    14101 acgcctcacg ggagctccaa gcggcgactg agatgtccta aatgcacagc gacggattcg
    14161 cgctatttag aaagagagag caatatttca agaatgcatg cgtcaatttt acgcagacta
    14221 tctttctagg gttaatctag ctgcatcagg atcatatcgt cgggtctttt ttccggctca
    14281 gtcatcgccc aagctggcgc tatctgggca tcggggagga agaagcccgt gccttttccc
    14341 gcgaggttga agcggcatgg aaagagtttg ccgaggatga ctgctgctgc attgacgttg
    14401 agcgaaaacg cacgtttacc atgatgattc gggaaggtgt ggccatgcac gcctttaacg
    14461 gtgaactgtt cgttcaggcc acctgggata ccagttcgtc gcggcttttc cggacacagt
    14521 tccggatggt cagcccgaag cgcatcagca acccgaacaa taccggcgac agccggaact
    14581 gccgtgccgg tgtgcagatt aatgacagcg gtgcggcgct gggatattac gtcagcgagg
    14641 acgggtatcc tggctggatg ccgcagaaat ggacatggat accccgtgag ttacccggcg
    14701 ggcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
    14761 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
    14821 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
    14881 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
    14941 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
    15001 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
    15061 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
    15121 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
    15181 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
    15241 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
    15301 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
    15361 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
    15421 taactatcgt cttgagtcca ac
//
