LOCUS       sequence-461182-       14206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-inducible lentiviral HACE Editor as a fusion of AID cytidine 
            deaminase and PcrA helicase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14206)
  AUTHORS   Chen XD, Chen Z, Wythes G, Zhang Y, Orr BC, Sun G, Chao YK, Navarro 
            Torres A, Thao K, Vallurupalli M, Sun J, Borji M, Tkacik E, Chen H, 
            Bernstein BE, Chen F
  TITLE     Helicase-assisted continuous editing for programmable mutagenesis of
            endogenous genomes.
  JOURNAL   Science. 2024 Oct 11;386(6718):eadn5876. doi: 
            10.1126/science.adn5876. Epub 2024 Oct 11.
  PUBMED    39388570
REFERENCE   2  (bases 1 to 14206)
  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: "Science."; 
            date: "2024-10-11"; volume: "386(6718):eadn5876. doi"; pages: " 
            10.1126/science.adn5876. Epub 2024 Oct 11"
COMMENT     Sequence Label: tet-AID-PcrA M6-UGI-T2A-mTagBFP2-PGK-Puro-T2A-...
FEATURES             Location/Qualifiers
     source          1..14206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2635..2949
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    2643..2661
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2679..2697
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2714..2732
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2750..2768
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2786..2804
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2821..2839
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2857..2875
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2922..2946
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2971..2995
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      3005..3014
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5828..6076
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6089..6109
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6110..6163
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             6164..6874
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     protein_bind    complement(6889..6913)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        6955..7465
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7384..7402
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7475..8071
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7475..7494)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7971..7991
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             8072..8125
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(8126..8743,8744..8749,8750..8785,8786..8788,8789..8824,
                     8825..8827,8828..8863,8864..8872)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    8898..9486
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8951..8971)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9369..9380)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9489..9505)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9490..9506)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10011..10191
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10217..10234)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10223..10447
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10493..10921
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10580..10599)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10790..10811
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10930..10950)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10935..11264
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11115..11250
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11177..11196
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11312..11359
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11378..11752
                     /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    11882..12003
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11919..11938)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11973..11992
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12052..12068)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12052..12068)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12065..12087)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12076..12092
                     /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(12100..12106,12107..12124,12125..12130))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12145..12166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12283..12300)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12454..13042)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12534..12553)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13213..14004,14005..14073))
                     /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     13836..13855
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14074..14178)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta acagagatcc actttggccg cgaatcgata tgtcgagttt
     2641 actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg
     2701 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt
     2761 gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgagttta
     2821 tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta
     2881 tgtcgaggta ggcgtgtacg gtgggaggcc tatataagca gagctcgttt agtgaaccgt
     2941 cagatcgcct ggagaattgg ctaagcatca acaagtttgt acaaaaaaac gggatccacc
     3001 ggtcgccacc atggacagcc tcttgatgaa ccggagggag tttctttacc aattcaaaaa
     3061 tgtccgctgg gctaagggtc ggcgtgagac ctacctgtgc tacgtagtga agaggcgtga
     3121 cagtgctaca tccttttcac tggactttgg ttatcttcgc aataagaacg gctgccacgt
     3181 ggaattgctc ttcctccgct acatctcgga ctgggaccta gaccctggcc gctgctaccg
     3241 cgtcacctgg ttcatctcct ggagcccctg ctacgactgt gcccgacatg tggccgactt
     3301 tctgcgaggg aaccccaacc tcagtctgag gatcttcacc gcgcgcctct acttctgtga
     3361 ggaccgcaag gctgagcccg aggggctgcg gcggctgcac cgcgccgggg tgcaaatagc
     3421 catcatgacc ttcaaagatt atttttactg ctggaatact tttgtagaaa accacggaag
     3481 aactttcaaa gcctgggaag ggctgcatga aaattcagtt cgtctctcca gacagcttcg
     3541 gcgcatcctt ttgcccctgt atgaggttga tgacttacga gacgcatttc gtactagcgg
     3601 cagcgagact cccgggacct cagagtccgc cacacccgaa agtatgaact tcctgtctga
     3661 gcaactgctg gcacacctga acaaggagca gcaagaagct gtgcggacca ccgagggacc
     3721 tctgctgatc atggccggcg ctggaagcgg aaaaacaaga gtgctgacac accgcatcgc
     3781 ctacctgatg gctgagaagc acgtggcccc ttggaacatc ctggccatta cctttacaaa
     3841 caaggccgct agagagatga gagaaagagt gcagagcctc ctgggaggcg ccgccgagga
     3901 cgtgtggatc agcaccttcg ccagcatggc cgtgcggatc ctgagaagag atatcgacag
     3961 aatcggcatc aaccggaact tcagcatcct ggacccaaca gaccagctga gcgtgatgaa
     4021 aaccatcctg aaagaaaaga acatcgaccc caagaaattc gagcctagaa caatcctggg
     4081 cacaatcagc gccgccaaga atgaactgct gcctccagaa cagtttgcga agcgggcctc
     4141 cacctactat gagaaagtgg tgtctgacgt ctaccaggag tatcagcagc ggctgctcag
     4201 gtgtcacagc cttgacttcg atgatctgat catgaccaca atccagctgt ttgaccgggt
     4261 ccccgacgtg ctccactact accaatacaa gtttcagtac atccacatcg atgagtacca
     4321 ggacacaaac agagcccaat acaccctggt gaaaaagctg gctgagcggt tccagaacat
     4381 cgccgccgtg ggcgacgccg atcagtctat ctacagatgg cggggcgccg acatccagaa
     4441 catcctgagc ttcgaaagag attaccccaa cgccaaagtg atcctgctgg agcaaaatta
     4501 ccggagcacg aagcgcatcc tgcaggccgc aaacgaggtg atcgagcaca acgtgaacag
     4561 aaagcctaag cggatctgga ccgagaatcc tgagggcaag cccatcctgt actacgaggc
     4621 catgaacgaa gccgacgagg cccagttcgt ggccggcaga atcagagagg ccgtggagcg
     4681 cggcgagaga agataccgag acttcgccgt gctgtacaga accaatgccc agtccagagt
     4741 catggaagag atgctgctga aggccaacat cccttaccag atagtgggcg gcgtgaagtt
     4801 ctacgacaga aaggaaatca aggacatcct ggcttatctg cgggtcatcg caaacccaga
     4861 cgacgactgc agcctgctga gaattatcaa cgttcctaaa agaggcatcg gagcctctac
     4921 catcgacaag ctggtccggt acgccgccga ccacgagctg agcctgttcg aggccctggg
     4981 cgagctggag atgatcggtc ttggcgccaa ggccgctggg gccctggccg ccttcagaag
     5041 ccagctggaa cagtggacac agctgcagga gtacgtgagc gtgaccgaac tggtggaaga
     5101 ggtgctggac aagtctggct accgggaaat gctgaaggcc gaaagaacca tcgaagccca
     5161 gagcagactg gagaacctgg atgagttcct gagcgtgacc aagcatttcg aaaacgtgtc
     5221 cgacgacaag agcctgatcg ccttcctgac cgatctggct ctgatcagcg acctggacga
     5281 gctggatggc accgagcagg ccgccgaggg cgacgccgtg atgctgatga ccctgcacgc
     5341 cgccaagggc ctggaattcc ccgtggtctt tctgataggc atggaagaag gcatcttccc
     5401 tcacaataga agcctggagg atgacgatga gatggaagag gagagaagac tggcctacgt
     5461 gggcatcacc agagctgaag aggaactggt gctgaccagc gcccagatgc ggaccctgtt
     5521 cggcaacatt cagatggacc ctcctagccg gttccttaac gagatccccg cccatctgct
     5581 tgagacagcc agcagacggc aggccggagc ttccaggcct gccgtgagca gaccccaggc
     5641 ctctggcgct gtgggcagct ggaaggtggg cgatcgggca aatcacagaa agtggggcat
     5701 cggaaccgtg gtttccgtgc ggggcggcgg agatgatcag gaactggaca tcgcttttcc
     5761 atctcctatc ggcattaaga gactgctagc taaattcgcc cctatcgaga aggtgtctgg
     5821 tggttctact aatctgtcag atattattga aaaggagacc ggtaagcaac tggttatcca
     5881 ggaatccatc ctcatgctcc cagaggaggt ggaagaagtc attgggaaca agccggaaag
     5941 cgatatactc gtgcacaccg cctacgacga gagcaccgac gagaatgtca tgcttctgac
     6001 tagcgacgcc cctgaataca agccttgggc tctggtcata caggatagca acggtgagaa
     6061 caagattaag atgctctctg gtggttctcc caagaagaag aggaaagtcg agggcagagg
     6121 aagtcttcta acatgcggtg acgtggagga gaatcccggc cctatggtgt ctaagggcga
     6181 agagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tggacaacca
     6241 tcacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag
     6301 aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctactagctt
     6361 cctctacggc agcaagacct tcatcaacca cacccagggc atccccgact tcttcaagca
     6421 gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct
     6481 gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag
     6541 aggggtgaac ttcacatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc
     6601 cttcaccgag acgctgtacc ccgctgacgg cggcctggaa ggcagaaacg acatggccct
     6661 gaagctcgtg ggcgggagcc atctgatcgc aaacgccaag accacatata gatccaagaa
     6721 acccgctaag aacctcaaga tgcctggcgt ctactatgtg gactacagac tggaaagaat
     6781 caaggaggcc aacaacgaga cctacgtcga gcagcacgag gtggcagtgg ccagatactg
     6841 cgacctccct agcaaactgg ggcacaagct taattaaagc ggccgcttcc ccagctttct
     6901 tgtacaaagt ggtttagtaa tgaaccggtc caccaccacc accactaagg atccggggtt
     6961 ggggttgcgc cttttccaag gcagccctgg gtttgcgcag ggacgcggct gctctgggcg
     7021 tggttccggg aaacgcagcg gcgccgaccc tgggtctcgc acattcttca cgtccgttcg
     7081 cagcgtcacc cggatcttcg ccgctaccct tgtgggcccc ccggcgacgc ttcctgctcc
     7141 gcccctaagt cgggaaggtt ccttgcggtt cgcggcgtgc cggacgtgac aaacggaagc
     7201 cgcacgtctc actagtaccc tcgcagacgg acagcgccag ggagcaatgg cagcgcgccg
     7261 accgcgatgg gctgtggcca atagcggctg ctcagcaggg cgcgccgaga gcagcggccg
     7321 ggaaggggcg gtgcgggagg cggggtgtgg ggcggtagtg tgggccctgt tcctgcccgc
     7381 gcggtgttcc gcattctgca agcctccgga gcgcacgtcg gcagtcggct ccctcgttga
     7441 ccgaatcacc gacctctctc cccagcaatt caccatgacc gagtacaagc ccacggtgcg
     7501 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     7561 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     7621 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     7681 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     7741 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     7801 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     7861 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     7921 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     7981 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     8041 cacctggtgc atgacccgca agcccggtgc cgaaggtaga ggttctctcc tcacttgtgg
     8101 tgatgttgaa gaaaaccctg gtccaatgtc tagactggac aagagcaaag tcataaacgg
     8161 agctctggaa ttactcaatg gtgtcggtat cgaaggcctg acgacaagga aactcgctca
     8221 aaagctggga gttgagcagc ctaccctgta ctggcacgtg aagaacaagc gggccctgct
     8281 cgatgccctg ccaatcgaga tgctggacag gcatcatacc cacttctgcc ccctggaagg
     8341 cgagtcatgg caagactttc tgcggaacaa cgccaagtca taccgctgtg ctctcctctc
     8401 acatcgcgac ggggctaaag tgcatctcgg cacccgccca acagagaaac agtacgaaac
     8461 cctggaaaat cagctcgcgt tcctgtgtca gcaaggcttc tccctggaga acgcactgta
     8521 cgctctgtcc gccgtgggcc actttacact gggctgcgta ttggaggaac aggagcatca
     8581 agtagcaaaa gaggaaagag agacacctac caccgattct atgcccccac ttctgagaca
     8641 agcaattgag ctgttcgacc ggcagggagc cgaacctgcc ttccttttcg gcctggaact
     8701 aatcatatgt ggcctggaga aacagctaaa gtgcgaaagc ggcgggccga ccgacgccct
     8761 tgacgatttt gacttagaca tgctcccagc cgatgccctt gacgactttg accttgatat
     8821 gctgcctgct gacgctcttg acgattttga ccttgacatg ctccccgggt aaagaattcg
     8881 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     8941 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     9001 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     9061 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     9121 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     9181 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     9241 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     9301 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     9361 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     9421 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     9481 ccccgcatcg ataccgtcga cctcgagacc tagaaaaaca tggagcaatc acaagtagca
     9541 atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg
     9601 gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     9661 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
     9721 aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat tggcagaact
     9781 acacaccagg gccagggatc agatatccac tgacctttgg atggtgctac aagctagtac
     9841 cagttgagca agagaaggta gaagaagcca atgaaggaga gaacacccgc ttgttacacc
     9901 ctgtgagcct gcatgggatg gatgacccgg agagagaagt attagagtgg aggtttgaca
     9961 gccgcctagc atttcatcac atggcccgag agctgcatcc ggactgtact gggtctctct
    10021 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
    10081 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
    10141 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agggcccgtt
    10201 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
    10261 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
    10321 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
    10381 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
    10441 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
    10501 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
    10561 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
    10621 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
    10681 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
    10741 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
    10801 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
    10861 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
    10921 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
    10981 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
    11041 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
    11101 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
    11161 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
    11221 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
    11281 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat
    11341 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
    11401 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
    11461 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
    11521 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
    11581 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
    11641 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
    11701 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct
    11761 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
    11821 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
    11881 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
    11941 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
    12001 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
    12061 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
    12121 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
    12181 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
    12241 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
    12301 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
    12361 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
    12421 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
    12481 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
    12541 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
    12601 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
    12661 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
    12721 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
    12781 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
    12841 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
    12901 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
    12961 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
    13021 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
    13081 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
    13141 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
    13201 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
    13261 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
    13321 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
    13381 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
    13441 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
    13501 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
    13561 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
    13621 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
    13681 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
    13741 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
    13801 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
    13861 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
    13921 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
    13981 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
    14041 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
    14101 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
    14161 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgac
//
