LOCUS       sequence-467753-       13259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Adenosine base editor 8e* with SpG nicking Cas9 makes A > G edits.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13259)
  AUTHORS   Sokirniy I, Inam H, Tomaszkiewicz M, Reynolds J, McCandlish D, 
            Pritchard J
  TITLE     A side-by-side comparison of variant function measurements using 
            deep mutational scanning and base editing.
  JOURNAL   Nucleic Acids Res. 2025 Jul 19;53(14):gkaf738. doi: 
            10.1093/nar/gkaf738.
  PUBMED    40744495
REFERENCE   2  (bases 1 to 13259)
  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: "Nucleic 
            Acids Res."; date: "2025-07-19"; volume: "53(14):gkaf738. doi"; 
            pages: " 10.1093/nar/gkaf738"
COMMENT     Sequence Label: ABE8e SpG Puro
FEATURES             Location/Qualifiers
     source          1..13259
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1040..1161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1051..1070)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1105..1124
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1291..1665)
                     /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"
     promoter        complement(1684..1731)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(1779..2108)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1793..1928
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1847..1866)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2093..2113
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2122..2550)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2232..2253)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2444..2463
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2596..2820
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2809..2826
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             2852..3085
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    3157..3745
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3263..3274
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3672..3692
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3761..4357)
                     /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     complement(3844..3864)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             complement(4358..4414)
                     /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             complement(4424..4444)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(4487..8587)
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFLWPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAKQLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(9182..9202)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(9252..9463)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     primer_bind     complement(9468..9488)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    9539..9656
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(9954..9998)
                     /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    10183..10416
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    10909..11034
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             11081..11261
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(11279..11477)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(11304..11324)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        11479..11858
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     12030..12049
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        12124..12228
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(12229..12297,12298..13089)
                     /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     complement(12447..12466)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
     1021 cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1081 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1141 ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg
     1201 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
     1261 ggcggtggaa tcgaaatctc gtagcacgtg tcagtcctgc tcctcggcca cgaagtgcac
     1321 gcagttgccg gccgggtcgc gcagggcgaa ctcccgcccc cacggctgct cgccgatctc
     1381 ggtcatggcc ggcccggagg cgtcccggaa gttcgtggac acgacctccg accactcggc
     1441 gtacagctcg tccaggccgc gcacccacac ccaggccagg gtgttgtccg gcaccacctg
     1501 gtcctggacc gcgctgatga acagggtcac gtcgtcccgg accacaccgg cgaagtcgtc
     1561 ctccacgaag tcccgggaga acccgagccg gtcggtccag aactcgaccg ctccggcgac
     1621 gtcgcgcgcg gtgagcaccg gaacggcact ggtcaacttg gccatggttt agttcctcac
     1681 cttgtcgtat tatactatgc cgatatacta tgccgatgat taattgtcaa cacgtgctga
     1741 tcagatccga aaatggatat acaagctccc gggagctttt tgcaaaagcc taggcctcca
     1801 aaaaagcctc ctcactactt ctggaatagc tcagaggcag aggcggcctc ggcctctgca
     1861 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
     1921 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
     1981 tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg
     2041 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
     2101 tgacacacat tccacagaat taattcgcgt taaatttttg ttaaatcagc tcatttttta
     2161 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     2221 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     2281 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     2341 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     2401 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     2461 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     2521 ccgcgcttaa tgcgccgcta cagggcgcgt ggggataccc cctagagccc cagctggttc
     2581 tttccgcctc agaagccata gagcccaccg catccccagc atgcctgcta ttgtcttccc
     2641 aatcctcccc cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca
     2701 gacaatgcga tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca
     2761 gggtcaagga aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag
     2821 tcgaggctga tcagcgggtt taaacgggcc ctgctagaga ttttccacac tgactaaaag
     2881 ggtctgaggg atctctagtt accagagtca cacaacagac gggcacacac tacttgaagc
     2941 actcaaggca agctttattg aggcttaagc agtgggttcc ctagttagcc agagagctcc
     3001 caggctcaga tctggtctaa ccagagagac ccagtacaag caaaaagcag atcttgtctt
     3061 cgttgggagt gaattagccc ttccagtccc cccttttctt ttaaaaagtg gctaagatct
     3121 acagctgcct tgtaagtcat tggtcttaaa gtcgacgcgg ggaggcggcc caaagggaga
     3181 tccgactcgt ctgagggcga aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc
     3241 cgcgggaagg aaggtccgct ggattgaggg ccgaagggac gtagcagaag gacgtcccgc
     3301 gcagaatcca ggtggcaaca caggcgagca gccaaggaaa ggacgatgat ttccccgaca
     3361 acaccacgga attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg
     3421 cggcgatgag ttccgccgtg gcaataggga gggggaaagc gaaagtcccg gaaaggagct
     3481 gacaggtggt ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca gtgcacacca
     3541 cgccacgttg cctgacaacg ggccacaact cctcataaag agacagcaac caggatttat
     3601 acaaggagga gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag
     3661 cagcgtatcc acatagcgta aaaggagcaa catagttaag aataccagtc aatctttcac
     3721 aaattttgta atccagaggt tgattgtcga cttaacgcgt tcaggcaccg ggcttgcggg
     3781 tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga
     3841 gccgctcgta gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg
     3901 cgcgctcggc cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt
     3961 ggtcgggcga gactccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg
     4021 ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc
     4081 gcgggccgat ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg
     4141 ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga
     4201 ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg acggtgtggc
     4261 gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt
     4321 cgcgggtggc gaggcgcacc gtgggcttgt actcggtcgg tccaggattc tcttcgacat
     4381 ctccggcttg tttcagcaga gagaagtttg ttgcgccagc gctgactttc ctcttcttct
     4441 tgggctcgaa ctcgctgccg tcggcggttc ttttggagcc tccgctgtca cctcccagct
     4501 gagacaggtc gatccgtgtc tcgtacaggc cggtgatgct ctggtggatc agggtggcgt
     4561 ccagcacctc tttggtgctt ctgtactgct tccggtcgat ggtggtgtca aagtacttga
     4621 aggcggcagg ggctcccaga ttggtcaggg taaacaggtg gatgatattc tcggcctgct
     4681 ctctgatggg cttatcccgg tgcttgttgt aggcggacag cactttgtcc agattagcgt
     4741 cggccaggat cactctcttg gagaactcgc tgatctgctc gatgatctcg tccaggtagt
     4801 gcttgtgctg ttccacaaac agctgtttct gctcattatc ctcgggggag cccttcagct
     4861 tctcatagtg gctggccagg tacaggaagt tcacatattt ggagggcagg gccagttcgt
     4921 ttcccttctg cagctgcttg gcagaggcca gcattctctt ccggccgttt tccagctcga
     4981 acagggagta cttaggcagc ttgatgatca ggtccttttt cacttctttg tagcccttgg
     5041 cttccagaaa gtcgatggga ttcttctcga agctgcttct ttccatgatg gtgatcccca
     5101 gcagctcttt cacactcttc agtttcttgg acttgccctt ttccactttg gccaccacca
     5161 gcacagaata ggccacggtg ggccacagga agccgccgta cttcttaggg tcccagtcct
     5221 tctttctggc gatcagctta tcgctgttcc tcttgggcag gatagactct ttgctgaagc
     5281 cgcctgtctg cacctcggtc tttttcacga tattcacttg gggcatgctc agcactttcc
     5341 gcacggtggc aaaatcccgg cccttatccc acacgatctc cccggtttcg ccgtttgtct
     5401 cgatcagagg ccgcttccgg atctcgccgt tggccagggt aatctcggtc ttgaaaaagt
     5461 tcatgatgtt gctgtagaag aagtacttgg cggtagcctt gccgatttcc tgctcgctct
     5521 tggcgatcat cttccgcacg tcgtacacct tgtagtcgcc gtacacgaac tcgctttcca
     5581 gcttagggta ctttttgatc agggcggttc ccacgacggc gttcaggtag gcgtcgtggg
     5641 cgtggtggta gttgttgatc tcgcgcactt tgtaaaactg gaaatccttc cggaaatcgg
     5701 acaccagctt ggacttcagg gtgatcactt tcacttcccg gatcagcttg tcattctcgt
     5761 cgtacttagt gttcatccgg gagtccagga tctgtgccac gtgctttgtg atctgccggg
     5821 tttccaccag ctgtctcttg atgaagccgg ccttatccag ttcgctcagg ccgcctctct
     5881 cggccttggt cagattgtcg aactttctct gggtaatcag cttggcgttc agcagctgcc
     5941 gccagtagtt cttcatcttc ttcacgacct cttcggaggg cacgttgtcg ctcttgcccc
     6001 ggttcttgtc gcttctggtc agcaccttgt tgtcgatgga gtcgtccttc agaaagctct
     6061 gaggcacgat atggtccaca tcgtagtcgg acagccggtt gatgtccagt tcctggtcca
     6121 cgtacatatc ccgcccattc tgcaggtagt acaggtacag cttctcgttc tgcagctggg
     6181 tgttttccac ggggtgttct ttcaggatct ggctgcccag ctctttgatg ccctcttcga
     6241 tccgcttcat tctctcgcgg ctgttcttct gtcccttctg ggtggtctgg ttctctctgg
     6301 ccatttcgat cacgatgttc tcgggcttgt gccggcccat cactttcacg agctcgtcca
     6361 ccaccttcac tgtctgcagg atgcccttct taatggcggg gctgccggcc agattggcaa
     6421 tgtgctcgtg caggctatcg ccctggccgg acacctgggc tttctggatg tcctctttaa
     6481 aggtcaggct gtcgtcgtgg atcagctgca tgaagtttct gttggcgaag ccgtcggact
     6541 tcaggaaatc caggattgtc ttgccggact gcttgtcccg gatgccgttg atcagcttcc
     6601 ggctcagcct gccccagccg gtgtatctcc gccgcttcag ctgcttcatc actttgtcgt
     6661 cgaacaggtg ggcataggtt ttcagccgtt cctcgatcat ctctctgtcc tcaaacagtg
     6721 tcagggtcag cacgatatct tccagaatgt cctcgttttc ctcattgtcc aggaagtcct
     6781 tgtccttgat aattttcagc agatcgtggt atgtgcccag ggaggcgttg aaccgatctt
     6841 ccacgccgga gatttccacg gagtcgaagc actcgatttt cttgaagtag tcctctttca
     6901 gctgcttcac ggtcactttc cggttggtct tgaacagcag gtccacgatg gcctttttct
     6961 gctcgccgct caggaaggcg ggctttctca ttccctcggt cacgtatttc actttggtca
     7021 gctcgttata cacggtgaag tactcgtaca gcaggctgtg cttgggcagc accttctcgt
     7081 tgggcaggtt cttatcgaag ttggtcatcc gctcgatgaa gctctgggcg gaagcgccct
     7141 tgtccaccac ttcctcgaag ttccaggggg tgatggtttc ctcgctcttt ctggtcatcc
     7201 aggcgaatct gctgtttccc ctggccagag ggcccacgta gtaggggatg cggaaggtca
     7261 ggatcttctc gatcttttcc cggttgtcct tcaggaatgg gtaaaaatct tcctgccgcc
     7321 gcagaatggc gtgcagctct cccaggtgga tctggtgggg gatgctgccg ttgtcgaagg
     7381 tccgctgctt ccgcagcagg tcctctctgt tcagcttcac gagcagttcc tcggtgccgt
     7441 ccatcttttc caggatgggc ttgatgaact tgtagaactc ttcctggctg gctccgccgt
     7501 caatgtagcc ggcgtagccg ttcttgctct ggtcgaagaa aatctctttg tacttctcag
     7561 gcagctgctg ccgcacgaga gctttcagca gggtcaggtc ctggtggtgc tcgtcgtatc
     7621 tcttgatcat agaggcgctc aggggggcct tggtgatctc ggtgttcact ctcaggatgt
     7681 cgctcagcag gatggcgtcg gacaggttct tggcggccag aaacaggtcg gcgtactggt
     7741 cgccgatctg ggccagcagg ttgtccaggt cgtcgtcgta ggtgtccttg ctcagctgca
     7801 gtttggcatc ctcggccagg tcgaagttgc tcttgaagtt gggggtcagg cccaggctca
     7861 gggcaatcag gtttccgaac aggccattct tcttctcgcc gggcagctgg gcgatcagat
     7921 tttccagccg tctgctcttg ctcagtctgg cagacaggat ggccttggcg tccacgccgc
     7981 tggcgttgat ggggttttcc tcgaacagct ggttgtaggt ctgcaccagc tggatgaaca
     8041 gcttgtccac gtcgctgttg tcggggttca ggtcgccctc gatcaggaag tggccccgga
     8101 acttgatcat gtgggccagg gccagataga tcagccgcag gtcggccttg tcggtgctgt
     8161 ccaccagttt ctttctcagg tggtagatgg tggggtactt ctcgtggtag gccacctcgt
     8221 ccacgatgtt gccgaagatg gggtgccgct cgtgcttctt atcctcttcc accaggaagg
     8281 actcttccag tctgtggaag aagctgtcgt ccaccttggc catctcgttg ctgaagatct
     8341 cttgcagata gcagatccgg ttcttccgtc tggtgtatct tcttctggcg gttctcttca
     8401 gccgggtggc ctcggctgtt tcgccgctgt cgaacagcag ggctccgatc aggttcttct
     8461 tgatgctgtg ccggtcggtg ttgcccagca ccttgaattt cttgctgggc accttgtact
     8521 cgtcggtgat cacggcccag cccacagagt tggtgccgat ggccaggccg atgctgtact
     8581 tcttgtcgct gccgccgctg ctgccgccac tgctctctgg tgttgctgac tcgcttgttc
     8641 ctggtgtctc gctgccagag gatcctccgc tagatcctcc agagttgatg gagctctggg
     8701 ccttcttctg agcattgaac acctgtctag gcatccgata gaaatcgcac agcagggcgg
     8761 cacattcatc tgccaggatt ccctcggtaa tttcgacgcg gtgattcatg ccggggtagt
     8821 tcagcacgtt catcagggag cctgcggcgc ctcttttaga atttctccat ccaaacacca
     8881 cgcggccgat cctagagtgg atcatggcgc cggcgcacat cacgcaaggc tcgaatgtca
     8941 cgtacagggt ggcgtcaatc agtctgtagt tctgcatgac caggccgccc tgtctcaggg
     9001 ccataatttc ggcatgggct gttgggtcgt gcaggccgat ggctctgttc cagccctcgc
     9061 cgatcactct attgttcagc accagcacgg ctcccacagg cacctccctc tcatcccgtg
     9121 ccctcttggc cagggtcagg gcatgtctca tccagtactc gtgggaaaac tccacctcag
     9181 agactttccg cttcttcttt ggtgactcga actcgcttcc gtcggctgtc cgtttcatgg
     9241 tggcaccggt cctgtgttct ggcggcaaac ccgttgcgaa aaagaacgtt cacggcgact
     9301 actgcactta tatacggttc tcccccaccc tcgggaaaaa ggcggagcca gtacacgaca
     9361 tcactttccc agtttacccc gcgccacctt ctctaggcac cggatcaatt gccgacccct
     9421 ccccccaact tctcggggac tgtgggcgat gtgcgctctg cccactgaat catgggaaat
     9481 aggccctcgg taccttaatt aaccaaactg gatctctgct gtccctgtaa taaacccgaa
     9541 aattttgaat ttttgtaatt tgtttttgta attctttagt ttgtatgtct gttgctatta
     9601 tgtctactat tctttcccct gcactgtacc ccccaatccc cccttttctt ttaaaattgt
     9661 ggatgaatac tgccatttgt ctgcagaatt ggcgcacgca gtgccgatcc gttcactaat
     9721 cgaatggatc tgtctctgtc tctctctcca ccttcttctt ctattccttc gggcctgtcg
     9781 ggtcccctcg gggttgggag gtgggtctga aacgataatg gtgaatatcc ctgcctaact
     9841 ctattcacta tagaaagtac agcaaaaact attcttaaac ctaccaagcc tcctactatc
     9901 attatgaata attttatata ccacagccaa tttgttatgt taaaccaatt ccacaaactt
     9961 gcccatttat ctaattccaa taattcttgt tcattctttt cttgctggtt ttgcgattct
    10021 tcaattaagg agtgtattaa gcttgtgtaa ttgttaattt ctctgtccca ctccatccag
    10081 gtcgtgtgat tccaaatctg ttccagagat ttattactcc aactagcatt ccaaggcaca
    10141 gcagtggtgc aaatgagttt tccagagcaa ccccaaatcc ccaggagctg ttgatccttt
    10201 aggtatcttt ccacagccag gattcttgcc tggagctgct tgatgcccca gactgtgagt
    10261 tgcaacagat gctgttgcgc ctcaatagcc ctcagcaaat tgttctgctg ctgcactata
    10321 ccagacaata attgtctggc ctgtaccgtc agcgtcattg acgctgcgcc catagtgctt
    10381 cctgctgctc ccaagaaccc aaggaacaaa gctcctattc ccactgctct tttttctctc
    10441 tgcaccactc ttctctttgc cttggtgggt gctactccta atggttcaat ttttactact
    10501 ttatatttat ataattcact tctccaattg tccctcatat ctcctcctcc aggtctgaag
    10561 atcagcggcc gcttgctgtg cggtggtctt acttttgttt tgctcttcct ctatcttgtc
    10621 taaagcttcc ttggtgtctt ttatctctat cctttgatgc acacaataga gggttgctac
    10681 tgtattatat aatgatctaa gttcttctga tcctgtctga agggatggtt gtagctgtcc
    10741 cagtatttgt ctacagcctt ctgatgtttc taacaggcca ggattaactg cgaatcgttc
    10801 tagctccctg cttgcccata ctatatgttt taatttatat tttttctttc cccctggcct
    10861 taaccgaatt ttttcccatc gcgatctaat tctcccccgc ttaatactga cgctctcgca
    10921 cccatctctc tccttctagc ctccgctagt caaaattttt ggcgtactca ccagtcgccg
    10981 cccctcgcct cttgccgtgc gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc
    11041 tggtttccct ttcgctttca agtccctgtt cgggcgccac tgctagagat tttccacact
    11101 gactaaaagg gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact
    11161 acttgaagca ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca
    11221 gagagctccc aggctcagat ctggtctaac cagagagacc cagtacaggc aaaacgcgct
    11281 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
    11341 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
    11401 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
    11461 ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt
    11521 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
    11581 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
    11641 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
    11701 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
    11761 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
    11821 aattgattac tattaataac tagtcaataa tcaatgtcaa cgcgtatatc tggcccgtac
    11881 atcgcgaagc agcgcaaaac gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa
    11941 gccttgcctt gttgtagctt aaattttgct cgcgcactac tcagcgacct ccaacacaca
    12001 agcagggagc agatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
    12061 ccatagggga tcgggagatc tcccgatccg tcgacgtcag gtggcacttt tcggggaaat
    12121 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
    12181 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
    12241 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
    12301 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
    12361 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
    12421 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
    12481 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
    12541 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
    12601 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
    12661 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
    12721 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
    12781 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
    12841 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
    12901 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
    12961 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
    13021 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
    13081 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
    13141 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
    13201 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttc
//
