LOCUS       sequence-424356-       12282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for VAPA clearance to acidic lysosomes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12282)
  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 12282)
  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"; 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-VAPA
FEATURES             Location/Qualifiers
     source          1..12282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   1069..1103
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    1437..1470
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        1620..1999
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2000..2203
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2154..2174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2200..2224
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3388..4383
                     /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    4419..4553
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4456..4475)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4510..4529
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    4582..4615
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5257..6435
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5488..6426
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     6383..6403
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    6496..6520
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(6529..6531,6532..6534,6535..7194)
                     /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(8118..8142)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             8195..8236
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(8274..8291)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8280..8504
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   9324..9386
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9424..9443)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9425..9443)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9450..9467)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9450..9466)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9459..9481)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             9604..9906
                     /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     9785..9804
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             10255..11049
                     /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             10255..11049
                     /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(10309..10328)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     10919..10938
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(11299..12090,12091..12159))
                     /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     11922..11941
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctca gaattaaccc
     1021 tcactaaagg gactagtcct gcaggtttaa acgaattcgc ccttttaacc ctagaaagat
     1081 agtctgcgta aaattgacgc atgcattctt gaaatattgc tctctctttc taaatagcgc
     1141 gaatccgtcg ctgtgcattt aggacatctc agtcgccgct tggagctccc gtgaggcgtg
     1201 cttgtcaatg cggtaagtgt cactgatttt gaactataac gaccgcgtga gtcaaaatga
     1261 cgcatgatta tcttttacgt gacttttaag atttaactca tacgataatt atattgttat
     1321 ttcatgttct acttacgtga taacttatta tatatatatt ttcttgttat agatatcaac
     1381 tagaatgcta gacctttcgt cttcaagaat tccgatcata ttcaataacc cttaatataa
     1441 cttcgtataa tgtatgctat acgaagttat taggtctgaa gaggagttta cgtccagcca
     1501 agcttaggat cccgggtacc gatggccggg cctcgaggag cttggcccat tgcatacgtt
     1561 gtatccatat cataatatgt acatttatat tggctcatgt ccaacattac cgccatgttg
     1621 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     1681 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     1741 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     1801 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     1861 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     1921 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     1981 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     2041 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     2101 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     2161 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
     2221 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc
     2281 cagcctcccc ggctagccgc cgccaccatg aaaaagcctg aactcaccgc gacgtctgtc
     2341 gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc
     2401 gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat
     2461 agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg
     2521 ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc
     2581 tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt
     2641 ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc
     2701 gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata
     2761 tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt
     2821 gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc
     2881 cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata
     2941 acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac
     3001 atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg
     3061 aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt
     3121 gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt
     3181 cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc
     3241 agaagcgcgg ccgtctggac cgatggctgt gtagaagtcg cgtctgcgtt cgaccaggct
     3301 gcgcgttctc gcggccatag caaccgacgt acggcgttgc gccctcgccg gcagcaagaa
     3361 gccacggaag tccgcccgga gcagaaaatg cccacgctac tgcgggttta tatagacggt
     3421 ccccacggga tggggaaaac caccaccacg caactgctgg tggccctggg ttcgcgcgac
     3481 gatatcgtct acgtacccga gccgatgact tactggcggg tgctgggggc ttccgagaca
     3541 atcgcgaaca tctacaccac acaacaccgc ctcgaccagg gtgagatatc ggccggggac
     3601 gcggcggtgg taatgacaag cgcccagata acaatgggca tgccttatgc cgtgaccgac
     3661 gccgttctgg ctcctcatat cgggggggag gctgggagct cacatgcccc gcccccggcc
     3721 ctcaccctca tcttcgaccg ccatcccatc gccgccctcc tgtgctaccc ggccgcgcgg
     3781 taccttatgg gcagcatgac cccccaggcc gtgctggcgt tcgtggccct catcccgccg
     3841 accttgcccg gcaccaacat cgtgcttggg gcccttccgg aggacagaca catcgaccgc
     3901 ctggccaaac gccagcgccc cggcgagcgg ctggacctgg ctatgctggc tgcgattcgc
     3961 cgcgtttacg ggctacttgc caatacggtg cggtatctgc agtgcggcgg gtcgtggcgg
     4021 gaggactggg gacagctttc ggggacggcc gtgccgcccc agggtgccga gccccagagc
     4081 aacgcgggcc cacgacccca tatcggggac acgttattta ccctgtttcg ggcccccgag
     4141 ttgctggccc ccaacggcga cctgtataac gtgtttgcct gggccttgga cgtcttggcc
     4201 aaacgcctcc gttccatgca cgtctttatc ctggattacg accaatcgcc cgccggctgc
     4261 cgggacgccc tgctgcaact tacctccggg atggtccaga cccacgtcac cacccccggc
     4321 tccataccga cgatatgcga cctggcgcgc acgtttgccc gggagatggg ggaggctaac
     4381 tgaagcttcc cgggggtacc gaattcgtcg acagatctaa cttgtttatt gcagcttata
     4441 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4501 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcatcaagc
     4561 ttaggatccg gaacccttaa tataacttcg tataatgtat gctatacgaa gttattaggt
     4621 ccctcgaaga ggttcactgg cgcgttggat ccccgggtac cgagttggga gctcacgggg
     4681 acagcccccc cccaaagccc ccagggatgt aattacgtcc ctcccccgct agggggcagc
     4741 agcgagccgc ccggggctcc gctccggtcc ggcgctcccc ccgcatcccc gagccggcag
     4801 cgtgcgggga cagcccgggc acggggaagg tggcacggga tcgctttcct ctgaacgctt
     4861 ctcgctgctc tttgagcctg cagacacctg gggggatacg gggaaaaagc tttaggctga
     4921 aagagagatt tagaatgaca gtctagtggg agctcacggg gacagccccc ccccaaagcc
     4981 cccagggatg taattacgtc cctcccccgc tagggggcag cagcgagccg cccggggctc
     5041 cgctccggtc cggcgctccc cccgcatccc cgagccggca gcgtgcgggg acagcccggg
     5101 cacggggaag gtggcacggg atcgctttcc tctgaacgct tctcgctgct ctttgagcct
     5161 gcagacacct ggggggatac ggggaaaaag ctttaggctg aaagagagat ttagaatgac
     5221 agaactcgat ttcattgcag actggcgcgc ccgtgaggct ccggtgcccg tcagtgggca
     5281 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt
     5341 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
     5401 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
     5461 cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc
     5521 ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg
     5581 attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa
     5641 ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg
     5701 cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa
     5761 aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc
     5821 caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg
     5881 tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg
     5941 gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc
     6001 ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg
     6061 cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg
     6121 ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac
     6181 tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg
     6241 tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg
     6301 gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt
     6361 gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca
     6421 tttcaggtgt cgtgaagaag cttggtaccg agctcggatc cactagtcca gtgtggtgga
     6481 attctgcaga tatcaacaag tttgtacaaa aaagcaggct gcggtaccat ggtgagcgtg
     6541 atcgccaagc agatgaccta caaggtgtac atgagcggca ccgtgaacgg ccactacttc
     6601 gaggtggagg gcgacggcaa gggcaagccc tacgagggcg agcagaccgt gaagctgacc
     6661 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgagccccca gctccagtac
     6721 ggcagcatcc ccttcaccaa gtaccccgag gacatccccg actacttcaa gcagagcttc
     6781 cccgagggct acacctggga gcgcagcatg aacttcgagg acggcgccgt gtgcaccgtg
     6841 agcaacgaca gcagcatcca gggcaactgc ttcatctaca acgtgaagat cagcggcgag
     6901 aacttccccc ccaacggccc cgtgatgcag aagaagaccc agggctggga gcccagcacc
     6961 gagcgcctgt tcgcccgcga cggaatgctg atcggcaacg actacatggc cctgaagctg
     7021 gagggcggcg gccactacct gtgcgagttc aagagcacct acaaggccaa gaagcccgtg
     7081 aggatgcccg gccgccacga gatcgaccgc aagctggacg tgaccagcca caaccgcgac
     7141 tacaccagcg tggagcagtg cgagatcgcc atcgcccgcc actccctgct gggcaagctt
     7201 ggaggatcag gaggatcaat gtctgcagca atggcgtccg cctcaggggc catggcgaag
     7261 cacgagcaga tcctggtcct cgatccgccc acagacctca aattcaaagg ccccttcaca
     7321 gatgtagtca ctacaaatct taaattgcga aatccatcgg atagaaaagt gtgtttcaaa
     7381 gtgaagacta cagcacctcg ccggtactgt gtgaggccca acagtggaat tattgaccca
     7441 gggtcaactg tgactgtttc agtaatgcta cagccctttg actatgatcc gaatgaaaag
     7501 agtaaacaca agtttatggt acagacaatt tttgctccac caaacacttc agatatggaa
     7561 gctgtgtgga aagaggcaaa acctgatgaa ttaatggatt ccaaattgag atgcgtattt
     7621 gaaatgccca atgaaaatga taaattgggt ataactccac cagggaatgc tccgactgtc
     7681 acttcaatga gcagcatcaa caacacagtt gcaacacctg ccagttatca cacgaaggat
     7741 gaccccaggg gactcagtgt gttgaaacag gagaaacaga agaatgatat ggaacctagc
     7801 aaagctgttc cactgaatgc atctaagcaa gatggaccta tgccaaaacc acacagtgtt
     7861 tcacttaatg ataccgaaac aaggaaacta atggaagagt gtaaaagact tcagggagaa
     7921 atgatgaagc tatcagaaga aaatcggcac ctgagagatg aaggtttaag gctcagaaag
     7981 gtagcacatt cggataaacc tggatcaacc tcaactgcat ccttcagaga taatgtcacc
     8041 agtcctcttc cttcacttct tgttgtaatt gcagccattt tcattggatt ctttctaggg
     8101 aaattcatct tgtagagacc cagctttctt gtacaaagtg gttgatatcc agcacagtgg
     8161 cggccgctcg agtctagagg gcccgcggtt cgaaggtaag cctatcccta accctctcct
     8221 cggtctcgat tctacgcgta ccggttaggg gcccgtttaa acccgctgat cagcctcgac
     8281 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     8341 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     8401 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     8461 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcggcgc gcctggatcc
     8521 ccgggtaccg agttgggagc tcacggggac agcccccccc caaagccccc agggatgtaa
     8581 ttacgtccct cccccgctag ggggcagcag cgagccgccc ggggctccgc tccggtccgg
     8641 cgctcccccc gcatccccga gccggcagcg tgcggggaca gcccgggcac ggggaaggtg
     8701 gcacgggatc gctttcctct gaacgcttct cgctgctctt tgagcctgca gacacctggg
     8761 gggatacggg gaaaaagctt taggctgaaa gagagattta gaatgacagt ctagtgggag
     8821 ctcacgggga cagccccccc ccaaagcccc cagggatgta attacgtccc tcccccgcta
     8881 gggggcagca gcgagccgcc cggggctccg ctccggtccg gcgctccccc cgcatccccg
     8941 agccggcagc gtgcggggac agcccgggca cggggaaggt ggcacgggat cgctttcctc
     9001 tgaacgcttc tcgctgctct ttgagcctgc agacacctgg ggggatacgg ggaaaaagct
     9061 ttaggctgaa agagagattt agaatgacag aactcgattt cattgcagac tggccggcca
     9121 ctagtacgcg ccggctcgac atactagtta aaagttttgt tactttatag aagaaatttt
     9181 gagtttttgt ttttttttaa taaataaata aacataaata aattgtttgt tgaatttatt
     9241 attagtatgt aagtgtaaat ataataaaac ttaatatcta ttcaaattaa taaataaacc
     9301 tcgatataca gaccgataaa acacatgcgt caattttacg catgattatc tttaacgtac
     9361 gtcacaatat gattatcttt ctagggttaa aagggcgaat tcgcggccgc taaattcaat
     9421 tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg
     9481 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     9541 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctatacgta
     9601 cggcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta
     9661 cagagtgata ttattgacac gccggggcga cggatggtga tccccctggc cagtgcacgt
     9721 ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc
     9781 tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg
     9841 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga
     9901 atataaatgt caggcatgag attatcaaaa aggatcttca cctagatcct tttcacgtag
     9961 aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg
    10021 acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga
    10081 tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc
    10141 tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc
    10201 tgatggcgca ggggatcaag ctctgatcaa gagacaggat gaggatcgtt tcgcatgatt
    10261 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
    10321 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
    10381 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
    10441 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
    10501 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
    10561 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
    10621 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
    10681 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
    10741 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
    10801 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
    10861 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
    10921 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
    10981 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
    11041 ttcttctgaa ttattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
    11101 tgcggtattt cacaccgcat caggtggcac ttttcgggga aatgtgcgcg gaacccctat
    11161 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagattat caaaaaggat
    11221 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
    11281 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
    11341 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
    11401 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
    11461 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
    11521 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
    11581 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
    11641 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
    11701 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
    11761 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
    11821 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
    11881 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
    11941 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
    12001 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
    12061 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
    12121 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
    12181 ttgaagcatt tatcagggtt attgtctcat gaccaaaatc ccttaacgtg agttttcgtt
    12241 ccactgagcg tcagaccccg tagaaaagat caaaggatct tc
//
