LOCUS       sequence-461364-       11945 bp    DNA     circular SYN 14-APR-2025
DEFINITION  PiggyBac Vector expressing KEIMA-REEP5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11945)
  AUTHORS   Hoyer MJ, Capitanio C, Smith IR, Paoli JC, Bieber A, Jiang Y, Paulo 
            JA, Gonzalez-Lozano MA, Baumeister W, Wilfling F, Schulman BA, 
            Harper JW
  TITLE     Combinatorial selective ER-phagy remodels the ER during 
            neurogenesis.
  JOURNAL   Nat Cell Biol. 2024 Mar;26(3):378-392. doi: 
            10.1038/s41556-024-01356-4. Epub 2024 Mar 1.
  PUBMED    38429475
REFERENCE   2  (bases 1 to 11945)
  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"; doi: 
            "10.1038/s41556-024-01356-4"; journalName: "Nat Cell Biol"; date: 
            "2024-03"; volume: "26"; issue: "3"; pages: "378-392"
COMMENT     Sequence Label: pAC150-KEIMA-REEP5
FEATURES             Location/Qualifiers
     source          1..11945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   299..333
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    667..700
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        850..1229
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1230..1433
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1384..1404
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1430..1454
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2618..3613
                     /codon_start=1
                     /product="truncated herpes simplex virus thymidine kinase 
                     (Salomon et al., 1995)"
                     /label=Delta-TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRV
                     LGASETIANIYTTQHRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGS
                     SHAPPPALTLIFDRHPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGAL
                     PEDRHIDRLAKRQRPGERLDLAMLAAIRRVYGLLANTVRYLQCGGSWREDWGQLSGTAV
                     PPQGAEPQSNAGPRPHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRSMHVFI
                     LDYDQSPAGCRDALLQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     polyA_signal    3649..3783
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3686..3705)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3740..3759
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    3812..3845
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4487..5665
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4718..5656
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5613..5633
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    5726..5750
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(5759..5761,5762..5764,5765..6424)
                     /codon_start=1
                     /product="humanized monomeric Keima-Red fluorescent 
                     protein"
                     /label=hmKeima-Red
                     /note="mammalian codon-optimized"
                     /translation="MVSVIAKQMTYKVYMSGTVNGHYFEVEGDGKGKPYEGEQTVKLTV
                     TKGGPLPFAWDILSPQLQYGSIPFTKYPEDIPDYFKQSFPEGYTWERSMNFEDGAVCTV
                     SNDSSIQGNCFIYNVKISGENFPPNGPVMQKKTQGWEPSTERLFARDGMLIGNDYMALK
                     LEGGGHYLCEFKSTYKAKKPVRMPGRHEIDRKLDVTSHNRDYTSVEQCEIAIARHSLLG
                     "
     protein_bind    complement(7011..7035)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             7088..7129
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(7167..7184)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7173..7397
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   8217..8279
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8317..8336)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8318..8336)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8343..8360)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8343..8359)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8352..8374)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             8497..8799
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     8678..8697
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             9148..9942
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             9148..9942
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(9202..9221)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9812..9831
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(10192..10983,10984..11052))
                     /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     10815..10834
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11176..11764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11665..11684
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11918..11935
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctca
      241 gaattaaccc tcactaaagg gactagtcct gcaggtttaa acgaattcgc ccttttaacc
      301 ctagaaagat agtctgcgta aaattgacgc atgcattctt gaaatattgc tctctctttc
      361 taaatagcgc gaatccgtcg ctgtgcattt aggacatctc agtcgccgct tggagctccc
      421 gtgaggcgtg cttgtcaatg cggtaagtgt cactgatttt gaactataac gaccgcgtga
      481 gtcaaaatga cgcatgatta tcttttacgt gacttttaag atttaactca tacgataatt
      541 atattgttat ttcatgttct acttacgtga taacttatta tatatatatt ttcttgttat
      601 agatatcaac tagaatgcta gacctttcgt cttcaagaat tccgatcata ttcaataacc
      661 cttaatataa cttcgtataa tgtatgctat acgaagttat taggtctgaa gaggagttta
      721 cgtccagcca agcttaggat cccgggtacc gatggccggg cctcgaggag cttggcccat
      781 tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt ccaacattac
      841 cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      901 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      961 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     1021 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     1081 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     1141 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1201 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     1261 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     1321 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     1381 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
     1441 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
     1501 gggaccgatc cagcctcccc ggctagccgc cgccaccatg aaaaagcctg aactcaccgc
     1561 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct
     1621 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct
     1681 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
     1741 atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac
     1801 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact
     1861 gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag
     1921 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg
     1981 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
     2041 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
     2101 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
     2161 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
     2221 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
     2281 cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg
     2341 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
     2401 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
     2461 aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtcg cgtctgcgtt
     2521 cgaccaggct gcgcgttctc gcggccatag caaccgacgt acggcgttgc gccctcgccg
     2581 gcagcaagaa gccacggaag tccgcccgga gcagaaaatg cccacgctac tgcgggttta
     2641 tatagacggt ccccacggga tggggaaaac caccaccacg caactgctgg tggccctggg
     2701 ttcgcgcgac gatatcgtct acgtacccga gccgatgact tactggcggg tgctgggggc
     2761 ttccgagaca atcgcgaaca tctacaccac acaacaccgc ctcgaccagg gtgagatatc
     2821 ggccggggac gcggcggtgg taatgacaag cgcccagata acaatgggca tgccttatgc
     2881 cgtgaccgac gccgttctgg ctcctcatat cgggggggag gctgggagct cacatgcccc
     2941 gcccccggcc ctcaccctca tcttcgaccg ccatcccatc gccgccctcc tgtgctaccc
     3001 ggccgcgcgg taccttatgg gcagcatgac cccccaggcc gtgctggcgt tcgtggccct
     3061 catcccgccg accttgcccg gcaccaacat cgtgcttggg gcccttccgg aggacagaca
     3121 catcgaccgc ctggccaaac gccagcgccc cggcgagcgg ctggacctgg ctatgctggc
     3181 tgcgattcgc cgcgtttacg ggctacttgc caatacggtg cggtatctgc agtgcggcgg
     3241 gtcgtggcgg gaggactggg gacagctttc ggggacggcc gtgccgcccc agggtgccga
     3301 gccccagagc aacgcgggcc cacgacccca tatcggggac acgttattta ccctgtttcg
     3361 ggcccccgag ttgctggccc ccaacggcga cctgtataac gtgtttgcct gggccttgga
     3421 cgtcttggcc aaacgcctcc gttccatgca cgtctttatc ctggattacg accaatcgcc
     3481 cgccggctgc cgggacgccc tgctgcaact tacctccggg atggtccaga cccacgtcac
     3541 cacccccggc tccataccga cgatatgcga cctggcgcgc acgtttgccc gggagatggg
     3601 ggaggctaac tgaagcttcc cgggggtacc gaattcgtcg acagatctaa cttgtttatt
     3661 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3721 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     3781 atcatcaagc ttaggatccg gaacccttaa tataacttcg tataatgtat gctatacgaa
     3841 gttattaggt ccctcgaaga ggttcactgg cgcgttggat ccccgggtac cgagttggga
     3901 gctcacgggg acagcccccc cccaaagccc ccagggatgt aattacgtcc ctcccccgct
     3961 agggggcagc agcgagccgc ccggggctcc gctccggtcc ggcgctcccc ccgcatcccc
     4021 gagccggcag cgtgcgggga cagcccgggc acggggaagg tggcacggga tcgctttcct
     4081 ctgaacgctt ctcgctgctc tttgagcctg cagacacctg gggggatacg gggaaaaagc
     4141 tttaggctga aagagagatt tagaatgaca gtctagtggg agctcacggg gacagccccc
     4201 ccccaaagcc cccagggatg taattacgtc cctcccccgc tagggggcag cagcgagccg
     4261 cccggggctc cgctccggtc cggcgctccc cccgcatccc cgagccggca gcgtgcgggg
     4321 acagcccggg cacggggaag gtggcacggg atcgctttcc tctgaacgct tctcgctgct
     4381 ctttgagcct gcagacacct ggggggatac ggggaaaaag ctttaggctg aaagagagat
     4441 ttagaatgac agaactcgat ttcattgcag actggcgcgc ccgtgaggct ccggtgcccg
     4501 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     4561 ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     4621 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     4681 cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg
     4741 cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct
     4801 gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc
     4861 ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg
     4921 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct
     4981 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt
     5041 aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg
     5101 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag
     5161 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc
     5221 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga
     5281 aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg
     5341 agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc
     5401 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt
     5461 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac
     5521 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt
     5581 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt
     5641 ttttcttcca tttcaggtgt cgtgaagaag cttggtaccg agctcggatc cactagtcca
     5701 gtgtggtgga attctgcaga tatcaacaag tttgtacaaa aaagcaggct gcggtaccat
     5761 ggtgagcgtg atcgccaagc agatgaccta caaggtgtac atgagcggca ccgtgaacgg
     5821 ccactacttc gaggtggagg gcgacggcaa gggcaagccc tacgagggcg agcagaccgt
     5881 gaagctgacc gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgagccccca
     5941 gctccagtac ggcagcatcc ccttcaccaa gtaccccgag gacatccccg actacttcaa
     6001 gcagagcttc cccgagggct acacctggga gcgcagcatg aacttcgagg acggcgccgt
     6061 gtgcaccgtg agcaacgaca gcagcatcca gggcaactgc ttcatctaca acgtgaagat
     6121 cagcggcgag aacttccccc ccaacggccc cgtgatgcag aagaagaccc agggctggga
     6181 gcccagcacc gagcgcctgt tcgcccgcga cggaatgctg atcggcaacg actacatggc
     6241 cctgaagctg gagggcggcg gccactacct gtgcgagttc aagagcacct acaaggccaa
     6301 gaagcccgtg aggatgcccg gccgccacga gatcgaccgc aagctggacg tgaccagcca
     6361 caaccgcgac tacaccagcg tggagcagtg cgagatcgcc atcgcccgcc actccctgct
     6421 gggcggaggc tcaggaggca tgtctgcggc catgagggag aggttcgacc ggttcctgca
     6481 cgagaagaac tgcatgactg accttctggc caagctcgag gccaaaaccg gcgtgaacag
     6541 gagcttcatc gctcttggtg tcatcggact ggtggccttg tacctggtgt tcggttatgg
     6601 agcctctctc ctctgcaacc tgataggatt tggctaccca gcctacatct caattaaagc
     6661 tatagagagt cccaacaaag aagatgatac ccagtggctg acctactggg tagtgtatgg
     6721 tgtgttcagc attgctgaat tcttctctga tatcttcctg tcatggttcc ccttctacta
     6781 catgctgaag tgtggcttcc tgttgtggtg catggccccg agcccttcta atggggctga
     6841 actgctctac aagcgcatca tccgtccttt cttcctgaag cacgagtccc agatggacag
     6901 tgtggtcaag gaccttaaag acaaggccaa agagactgca gatgccatca ctaaagaagc
     6961 gaagaaagct accgtgaatt tactgggtga agaaaagaag agcacctaag acccagcttt
     7021 cttgtacaaa gtggttgata tccagcacag tggcggccgc tcgagtctag agggcccgcg
     7081 gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtta
     7141 ggggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     7201 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     7261 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     7321 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     7381 tgcggtgggc tctatggcgg cgcgcctgga tccccgggta ccgagttggg agctcacggg
     7441 gacagccccc ccccaaagcc cccagggatg taattacgtc cctcccccgc tagggggcag
     7501 cagcgagccg cccggggctc cgctccggtc cggcgctccc cccgcatccc cgagccggca
     7561 gcgtgcgggg acagcccggg cacggggaag gtggcacggg atcgctttcc tctgaacgct
     7621 tctcgctgct ctttgagcct gcagacacct ggggggatac ggggaaaaag ctttaggctg
     7681 aaagagagat ttagaatgac agtctagtgg gagctcacgg ggacagcccc cccccaaagc
     7741 ccccagggat gtaattacgt ccctcccccg ctagggggca gcagcgagcc gcccggggct
     7801 ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc agcgtgcggg gacagcccgg
     7861 gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc ttctcgctgc tctttgagcc
     7921 tgcagacacc tggggggata cggggaaaaa gctttaggct gaaagagaga tttagaatga
     7981 cagaactcga tttcattgca gactggccgg ccactagtac gcgccggctc gacatactag
     8041 ttaaaagttt tgttacttta tagaagaaat tttgagtttt tgtttttttt taataaataa
     8101 ataaacataa ataaattgtt tgttgaattt attattagta tgtaagtgta aatataataa
     8161 aacttaatat ctattcaaat taataaataa acctcgatat acagaccgat aaaacacatg
     8221 cgtcaatttt acgcatgatt atctttaacg tacgtcacaa tatgattatc tttctagggt
     8281 taaaagggcg aattcgcggc cgctaaattc aattcgccct atagtgagtc gtattacaat
     8341 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     8401 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     8461 cgcccttccc aacagttgcg cagcctatac gtacggcagt ttaaggttta cacctataaa
     8521 agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga cacgccgggg
     8581 cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa
     8641 ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac cgatatggcc
     8701 agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg cgaaaatgac
     8761 atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggcat gagattatca
     8821 aaaaggatct tcacctagat ccttttcacg tagaaagcca gtccgcagaa acggtgctga
     8881 ccccggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga
     8941 aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt tttatggaca
     9001 gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa gccctgcaaa
     9061 gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc aagctctgat
     9121 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     9181 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     9241 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     9301 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     9361 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     9421 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     9481 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     9541 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     9601 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     9661 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     9721 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     9781 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     9841 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     9901 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gaattattaa cgcttacaat
     9961 ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catcaggtgg
    10021 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
    10081 tatgtatccg ctcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
    10141 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
    10201 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
    10261 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
    10321 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
    10381 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
    10441 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
    10501 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
    10561 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
    10621 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
    10681 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
    10741 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
    10801 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
    10861 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
    10921 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
    10981 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
    11041 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
    11101 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
    11161 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
    11221 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
    11281 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
    11341 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
    11401 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
    11461 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
    11521 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
    11581 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
    11641 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
    11701 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
    11761 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
    11821 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
    11881 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
    11941 ggaag
//
