LOCUS       sequence-332993-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Disrupt 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_3
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(54..73)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        245..624
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        626..824
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     779..799
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             842..1022
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1069..1194
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1687..1920
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2105..2149
                     /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    2447..2564
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2615..2863
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2615..2635
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2786..2805
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2885..2960
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3022..3233
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3252..3261
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3258..7361
                     /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             3258..7361
                     /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             7362..7409
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7410..7433
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7443..7499
                     /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             7500..8096
                     /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     7993..8013
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8112..8700
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8165..8185)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8583..8594)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8772..9005
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9031..9048)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9037..9261
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9307..9735
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9394..9413)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9604..9625
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9744..9764)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9749..10078
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9929..10064
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9991..10010
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10126..10173
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10192..10566
                     /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    10696..10817
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10733..10752)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10787..10806
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10866..10882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10866..10882)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10879..10901)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10890..10906
                     /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(10914..10920,10921..10938,10939..10944))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10959..10980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11097..11114)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11268..11856)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11348..11367)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12027..12818,12819..12887))
                     /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     12650..12669
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12888..12992)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
       61 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
      121 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
      181 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
      241 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
      301 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      361 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      421 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      481 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      541 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      601 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      661 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      721 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      781 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac
      841 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
      901 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
      961 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     1021 cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc
     1081 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg
     1141 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt
     1201 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa
     1261 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
     1321 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
     1381 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
     1441 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
     1501 caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg
     1561 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     1621 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     1681 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     1741 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     1801 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     1861 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     1921 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     1981 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     2041 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     2101 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     2161 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     2221 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     2281 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     2341 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc
     2401 actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag
     2461 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2521 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2581 acagcagaga tccagtttgg ttaattaagg taccgagggc ctatttccca tgattccttc
     2641 atatttgcat atacgataca aggctgttag agagataatt agaattaatt tgactgtaaa
     2701 cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc
     2761 agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt
     2821 cgatttcttg gctttatata tcttgtggaa aggacgaaac accggctcgc tacttaacta
     2881 acaggtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa
     2941 aagtggcacc gagtcggtgc ttttttgaat tcgctagcta ggtcttgaaa ggagtgggaa
     3001 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     3061 ggggaggggt cggcaattga tccggtgcct agagaaggtg gcgcggggta aactgggaaa
     3121 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     3181 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggaccggt
     3241 tctagagcgc tgccaccatg gacaagaagt acagcatcgg cctggacatc ggcaccaact
     3301 ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc
     3361 tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca
     3421 gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac
     3481 ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg
     3541 acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc
     3601 ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca
     3661 tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct
     3721 atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga
     3781 accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc
     3841 tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca
     3901 gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga
     3961 atggcctgtt cggaaacctg attgccctga gcctgggcct gacccccaac ttcaagagca
     4021 acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc
     4081 tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga
     4141 acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg
     4201 cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc
     4261 tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga
     4321 gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt
     4381 tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca
     4441 gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc
     4501 acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg
     4561 acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc
     4621 tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc
     4681 cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga
     4741 tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt
     4801 acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga
     4861 gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga
     4921 ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct
     4981 tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc
     5041 acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca
     5101 ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac
     5161 ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga
     5221 gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt
     5281 ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc
     5341 agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg
     5401 gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga
     5461 agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca
     5521 agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga
     5581 agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga
     5641 tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact
     5701 acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg
     5761 actacgatgt ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca
     5821 aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg
     5881 tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga
     5941 gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg
     6001 gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc
     6061 tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag
     6121 tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag
     6181 tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa
     6241 ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg
     6301 tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca
     6361 agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg
     6421 gcgagatccg gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg
     6481 ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg
     6541 tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga
     6601 acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg
     6661 acagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca
     6721 agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg
     6781 agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga
     6841 tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg
     6901 cctctgccgg cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact
     6961 tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga
     7021 aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg
     7081 agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca
     7141 acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc
     7201 tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga
     7261 ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc
     7321 tgtacgagac acggatcgac ctgtctcagc tgggaggcga caagcgacct gccgccacaa
     7381 agaaggctgg acaggctaag aagaagaaag attacaaaga cgatgacgat aagggatccg
     7441 gcgcaacaaa cttctctctg ctgaaacaag ccggagatgt cgaagagaat cctggaccga
     7501 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
     7561 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
     7621 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     7681 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     7741 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     7801 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     7861 cgtggttcct ggccaccgtc ggagtctcgc ccgaccacca gggcaagggt ctgggcagcg
     7921 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     7981 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     8041 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgaacgc
     8101 gttaagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     8161 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     8221 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     8281 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     8341 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     8401 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     8461 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     8521 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     8581 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     8641 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     8701 gtcgacttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     8761 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     8821 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     8881 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     8941 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     9001 tagcagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     9061 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     9121 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     9181 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     9241 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     9301 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     9361 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     9421 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     9481 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     9541 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     9601 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     9661 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     9721 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     9781 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     9841 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     9901 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     9961 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
    10021 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
    10081 gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca
    10141 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag
    10201 ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg
    10261 accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg
    10321 gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg
    10381 gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc
    10441 acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg
    10501 cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag
    10561 gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
    10621 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
    10681 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
    10741 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
    10801 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
    10861 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
    10921 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
    10981 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
    11041 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
    11101 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
    11161 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
    11221 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
    11281 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
    11341 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
    11401 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
    11461 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
    11521 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
    11581 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
    11641 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
    11701 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
    11761 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
    11821 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
    11881 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
    11941 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
    12001 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
    12061 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
    12121 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
    12181 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
    12241 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
    12301 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
    12361 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
    12421 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
    12481 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
    12541 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
    12601 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
    12661 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
    12721 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
    12781 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
    12841 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
    12901 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
    12961 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gcc
//
