LOCUS       sequence-332994-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Disrput ACO1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  AUTHORS   Terzi EM, Sviderskiy VO, Alvarez SW, Whiten GC, Possemato R
  TITLE     Iron-sulfur cluster deficiency can be sensed by IRP2 and regulates 
            iron homeostasis and sensitivity to ferroptosis independent of IRP1 
            and FBXL5.
  JOURNAL   Sci Adv. 2021 May 26;7(22). pii: 7/22/eabg4302. doi: 
            10.1126/sciadv.abg4302. Print 2021 May.
  PUBMED    34039609
REFERENCE   2  (bases 1 to 13013)
  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: "Sci Adv."; 
            date: "2021-05-26"; volume: "7(22). pii: 7/22/eabg4302. doi"; pages:
            " 10.1126/sciadv.abg4302. Print 2021 May"
COMMENT     Sequence Label: pLentiCRISPR V2 sgACO1_2
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(542..1333,1334..1402))
                     /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     1165..1184
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1403..1507)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1582..1601)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1773..2152
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2154..2352
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2307..2327
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2370..2550
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2597..2722
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3215..3448
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3633..3677
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    3975..4092
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4143..4391
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4143..4163
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4314..4333
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4413..4488
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4550..4761
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4780..4789
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4786..8889
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4786..8889
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8890..8937
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8938..8961
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8971..9027
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             9028..9624
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     9521..9541
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9640..10228
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9693..9713)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10111..10122)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10300..10533
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10559..10576)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10565..10789
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10835..11263
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10922..10941)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11132..11153
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11272..11292)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11277..11606
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11457..11592
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11519..11538
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11654..11701
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11720..12094
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    12224..12345
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12261..12280)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12315..12334
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12394..12410)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12394..12410)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12407..12429)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12418..12434
                     /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(12442..12448,12449..12466,12467..12472))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12487..12508
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12625..12642)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(12796..13013,1..371))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12876..12895)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
       61 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
      121 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
      181 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
      241 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
      301 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
      361 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
      421 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
      481 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
      541 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
      601 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
      661 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
      721 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
      781 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
      841 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
      901 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
      961 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     1021 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     1081 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     1141 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     1201 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     1261 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     1321 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     1381 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     1441 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     1501 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     1561 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     1621 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     1681 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     1741 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
     1801 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     1861 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     1921 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     1981 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     2041 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     2101 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     2161 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     2221 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     2281 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     2341 ctatataagc agcgcgtttt gcctgtactg ggtctctctg gttagaccag atctgagcct
     2401 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     2461 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     2521 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
     2581 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     2641 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     2701 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     2761 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     2821 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     2881 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     2941 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     3001 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     3061 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     3121 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     3181 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     3241 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     3301 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     3361 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     3421 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     3481 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     3541 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     3601 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     3661 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     3721 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     3781 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     3841 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     3901 gatccattcg attagtgaac ggatcggcac tgcgtgcgcc aattctgcag acaaatggca
     3961 gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     4021 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     4081 attcaaaatt ttcgggttta ttacagggac agcagagatc cagtttggtt aattaaggta
     4141 ccgagggcct atttcccatg attccttcat atttgcatat acgatacaag gctgttagag
     4201 agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta
     4261 gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc
     4321 atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag
     4381 gacgaaacac cgccactacc ccaacctggc gggttttaga gctagaaata gcaagttaaa
     4441 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttttgaattc
     4501 gctagctagg tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc
     4561 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgatc cggtgcctag
     4621 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     4681 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     4741 gggtttgccg ccagaacaca ggaccggttc tagagcgctg ccaccatgga caagaagtac
     4801 agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac
     4861 aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag
     4921 aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag
     4981 agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc
     5041 ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc
     5101 ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag
     5161 gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc
     5221 accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg
     5281 ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc
     5341 atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc
     5401 gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg
     5461 atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc
     5521 ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag
     5581 ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag
     5641 tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc
     5701 ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac
     5761 gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag
     5821 aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc
     5881 ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc
     5941 accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc
     6001 gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg
     6061 caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc
     6121 ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg
     6181 accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc
     6241 gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag
     6301 aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc
     6361 aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa
     6421 aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa
     6481 gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat
     6541 cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac
     6601 ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg
     6661 tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac
     6721 aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag
     6781 ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc
     6841 gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa
     6901 gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc
     6961 aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac
     7021 gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga
     7081 gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa
     7141 gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag
     7201 ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac
     7261 caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc
     7321 tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc
     7381 aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag
     7441 ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga
     7501 ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg
     7561 cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag
     7621 aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat
     7681 ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac
     7741 gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa
     7801 agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc
     7861 gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt
     7921 ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca
     7981 aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa
     8041 gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc
     8101 agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac
     8161 tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg
     8221 gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg
     8281 atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag
     8341 ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag
     8401 ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa
     8461 ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg
     8521 aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac
     8581 ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct
     8641 aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag
     8701 gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag
     8761 tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc
     8821 accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg
     8881 ggaggcgaca agcgacctgc cgccacaaag aaggctggac aggctaagaa gaagaaagat
     8941 tacaaagacg atgacgataa gggatccggc gcaacaaact tctctctgct gaaacaagcc
     9001 ggagatgtcg aagagaatcc tggaccgacc gagtacaagc ccacggtgcg cctcgccacc
     9061 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     9121 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     9181 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     9241 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     9301 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     9361 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg agtctcgccc
     9421 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     9481 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     9541 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     9601 atgacccgca agcccggtgc ctgaacgcgt taagtcgaca atcaacctct ggattacaaa
     9661 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     9721 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     9781 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     9841 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     9901 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     9961 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
    10021 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
    10081 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
    10141 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
    10201 cggatctccc tttgggccgc ctccccgcgt cgactttaag accaatgact tacaaggcag
    10261 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
    10321 aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
    10381 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
    10441 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
    10501 gaccctttta gtcagtgtgg aaaatctcta gcagggcccg tttaaacccg ctgatcagcc
    10561 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
    10621 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
    10681 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
    10741 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
    10801 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
    10861 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
    10921 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
    10981 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
    11041 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
    11101 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
    11161 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
    11221 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
    11281 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
    11341 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
    11401 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
    11461 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
    11521 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
    11581 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
    11641 ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
    11701 caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg
    11761 cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt
    11821 ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca
    11881 ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct
    11941 gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat
    12001 gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa
    12061 ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt tcgattccac
    12121 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
    12181 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
    12241 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
    12301 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
    12361 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
    12421 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
    12481 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
    12541 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
    12601 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
    12661 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
    12721 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
    12781 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
    12841 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
    12901 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
    12961 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tca
//
