LOCUS       sequence-332996-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Disrupt IREB2.
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 sgIREB2_2
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..32)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        204..583
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        585..783
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     738..758
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             801..981
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1028..1153
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1646..1879
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2064..2108
                     /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    2406..2523
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2574..2822
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2574..2594
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2745..2764
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2844..2919
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2981..3192
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3211..3220
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3217..7320
                     /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             3217..7320
                     /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             7321..7368
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7369..7392
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7402..7458
                     /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             7459..8055
                     /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     7952..7972
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8071..8659
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8124..8144)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8542..8553)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8731..8964
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8990..9007)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8996..9220
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9266..9694
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9353..9372)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9563..9584
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9703..9723)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9708..10037
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9888..10023
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9950..9969
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10085..10132
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10151..10525
                     /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    10655..10776
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10692..10711)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10746..10765
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10825..10841)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10825..10841)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10838..10860)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10849..10865
                     /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(10873..10879,10880..10897,10898..10903))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10918..10939
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11056..11073)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11227..11815)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11307..11326)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11986..12777,12778..12846))
                     /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     12609..12628
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12847..12951)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
       61 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
      121 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
      181 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
      241 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      301 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      361 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      421 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      481 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      541 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      601 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      661 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      721 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      781 cagcgcgttt tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct
      841 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
      901 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
      961 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
     1021 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
     1081 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
     1141 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
     1201 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
     1261 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
     1321 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     1381 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     1441 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga
     1501 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
     1561 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
     1621 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
     1681 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
     1741 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
     1801 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
     1861 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
     1921 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
     1981 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
     2041 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
     2101 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
     2161 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
     2221 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
     2281 ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc
     2341 gattagtgaa cggatcggca ctgcgtgcgc caattctgca gacaaatggc agtattcatc
     2401 cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac
     2461 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat
     2521 tttcgggttt attacaggga cagcagagat ccagtttggt taattaaggt accgagggcc
     2581 tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga gagataatta
     2641 gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat
     2701 aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta
     2761 ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca
     2821 ccgtgagcca ttccagttcc agggttttag agctagaaat agcaagttaa aataaggcta
     2881 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttgaatt cgctagctag
     2941 gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg cacatcgccc
     3001 acagtccccg agaagttggg gggaggggtc ggcaattgat ccggtgccta gagaaggtgg
     3061 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
     3121 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
     3181 gccagaacac aggaccggtt ctagagcgct gccaccatgg acaagaagta cagcatcggc
     3241 ctggacatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc
     3301 agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc
     3361 ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc
     3421 agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac
     3481 gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa
     3541 gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac
     3601 cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag
     3661 gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc
     3721 ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg
     3781 gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc
     3841 aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag
     3901 ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg
     3961 acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag
     4021 gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac
     4081 ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg
     4141 aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac
     4201 caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa
     4261 gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc
     4321 caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa
     4381 ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc
     4441 agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat
     4501 ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc
     4561 ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag
     4621 agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc
     4681 cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg
     4741 cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa
     4801 tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc
     4861 gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac
     4921 ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac
     4981 gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac
     5041 aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac
     5101 agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg
     5161 aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac
     5221 ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc
     5281 gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc
     5341 cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc
     5401 ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg
     5461 aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag
     5521 accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc
     5581 aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac
     5641 gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg
     5701 gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag
     5761 gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac
     5821 aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac
     5881 gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg
     5941 agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca
     6001 aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag
     6061 ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag
     6121 gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac
     6181 ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc
     6241 gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa
     6301 atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc
     6361 gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa
     6421 accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc
     6481 atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag
     6541 tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct
     6601 aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa
     6661 gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc
     6721 atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa
     6781 gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac
     6841 ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg
     6901 ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc
     6961 cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag
     7021 atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac
     7081 aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat
     7141 atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac
     7201 accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc
     7261 caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac
     7321 aagcgacctg ccgccacaaa gaaggctgga caggctaaga agaagaaaga ttacaaagac
     7381 gatgacgata agggatccgg cgcaacaaac ttctctctgc tgaaacaagc cggagatgtc
     7441 gaagagaatc ctggaccgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     7501 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     7561 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     7621 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     7681 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     7741 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     7801 caccggccca aggagcccgc gtggttcctg gccaccgtcg gagtctcgcc cgaccaccag
     7861 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     7921 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     7981 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     8041 aagcccggtg cctgaacgcg ttaagtcgac aatcaacctc tggattacaa aatttgtgaa
     8101 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     8161 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     8221 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     8281 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     8341 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     8401 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     8461 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     8521 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     8581 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     8641 ctttgggccg cctccccgcg tcgactttaa gaccaatgac ttacaaggca gctgtagatc
     8701 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
     8761 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     8821 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     8881 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     8941 agtcagtgtg gaaaatctct agcagggccc gtttaaaccc gctgatcagc ctcgactgtg
     9001 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     9061 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     9121 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     9181 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     9241 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     9301 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     9361 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     9421 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     9481 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     9541 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     9601 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     9661 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     9721 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     9781 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     9841 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     9901 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     9961 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
    10021 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
    10081 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
    10141 gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc
    10201 cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga
    10261 cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt
    10321 ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga
    10381 gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat
    10441 cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca
    10501 cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt
    10561 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
    10621 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
    10681 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
    10741 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
    10801 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
    10861 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
    10921 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
    10981 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
    11041 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
    11101 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
    11161 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
    11221 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
    11281 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
    11341 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
    11401 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
    11461 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
    11521 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
    11581 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
    11641 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
    11701 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
    11761 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
    11821 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
    11881 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
    11941 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
    12001 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
    12061 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
    12121 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
    12181 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
    12241 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
    12301 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
    12361 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
    12421 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
    12481 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
    12541 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
    12601 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
    12661 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
    12721 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
    12781 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
    12841 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
    12901 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
    12961 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atg
//
