LOCUS       sequence-416335-       10094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible lentiviral expression, TRE-RPS2-APOBEC-HA-P2A-mRuby; 
            PGK-puro-2A-rtTA (Ribo-STAMP, RPS2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10094)
  AUTHORS   Brannan KW, Chaim IA, Marina RJ, Yee BA, Kofman ER, Lorenz DA, 
            Jagannatha P, Dong KD, Madrigal AA, Underwood JG, Yeo GW
  TITLE     Robust single-cell discovery of RNA targets of RNA-binding proteins 
            and ribosomes.
  JOURNAL   Nat Methods. 2021 May;18(5):507-519. doi: 
            10.1038/s41592-021-01128-0. Epub 2021 May 7.
  PUBMED    33963355
REFERENCE   2  (bases 1 to 10094)
  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"; doi: 
            "10.1038/s41592-021-01128-0"; journalName: "Nat Methods"; date: 
            "2021-05"; volume: "18"; issue: "5"; pages: "507-519"
COMMENT     Sequence Label: pLIX403_RPS2_APOBEC_HA_P2A_mRuby_Capture1
FEATURES             Location/Qualifiers
     source          1..10094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(96..115)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      909..1497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1398..1417
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1651..1668
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1785..1806
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1821..1826,1827..1844,1845..1851)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1859..1875
                     /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     1864..1886
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1883..1899
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1883..1899
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1918..1938
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1920..1938
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1966..2192
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2193..2373
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2420..2545
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3038..3271
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3456..3500
                     /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    3713..3830
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3884..4198
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    3892..3910
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3928..3946
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3963..3981
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3999..4017
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4035..4053
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4070..4088
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4106..4124
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4171..4195
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    4219..4243
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5130..5154)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5183..5869
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             5876..5902
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             5876..5902
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5903..5959
                     /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             5963..6670
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTDI
                     ALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAVA
                     KYSNLGGGMDELYK"
     CDS             6681..6698
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        6708..7218
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7137..7155
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7228..7824
                     /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(7228..7247)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7724..7744
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             7825..7878
                     /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(7879..8496,8497..8502,8503..8538,8539..8541,8542..8577,
                     8578..8580,8581..8616,8617..8625)
                     /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"
     primer_bind     complement(8665..8685)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9083..9094)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9299..9532
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9604..9725
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9641..9660)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9695..9714
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9765..9900
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        9900..9971
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9972..10040,10041..10094,1..738)
                     /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"
ORIGIN
        1 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
       61 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      121 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      181 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      241 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
      301 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
      361 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
      421 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
      481 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
      541 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
      601 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
      661 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
      721 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
      781 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
      841 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
      901 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
      961 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1021 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     1081 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1141 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1201 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1261 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     1321 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     1381 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     1441 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     1501 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     1561 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     1621 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     1681 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     1741 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     1801 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     1861 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
     1921 attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa
     1981 tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga
     2041 aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa
     2101 ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata
     2161 ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag
     2221 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     2281 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     2341 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
     2401 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
     2461 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
     2521 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
     2581 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
     2641 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
     2701 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     2761 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     2821 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     2881 agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat
     2941 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
     3001 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
     3061 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac
     3121 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
     3181 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
     3241 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
     3301 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt
     3361 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
     3421 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
     3481 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
     3541 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
     3601 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
     3661 ccccgagggg acaattctcg acctcgagac aaatggcagt attcatccac aattttaaaa
     3721 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     3781 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     3841 acagggacag cagagatcca ctttggccgc gaatcgatat gtcgagttta ctccctatca
     3901 gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacga tgtcgagttt
     3961 actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg
     4021 tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttat ccctatcagt
     4081 gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgaggtag
     4141 gcgtgtacgg tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgcctg
     4201 gagaattggc tagcatcaac aagtttgtac aaaaaagcag gcttacacca tggcggatga
     4261 cgccggtgca gcgggggggc ccgggggccc tggtggccct gggatgggga accgcggtgg
     4321 cttccgcgga ggtttcggca gtggcatccg gggccggggt cgcggccgtg gacggggccg
     4381 gggccgaggc cgcggagctc gcggaggcaa ggccgaggat aaggagtgga tgcccgtcac
     4441 caagttgggc cgcttggtca aggacatgaa gatcaagtcc ctggaggaga tctatctctt
     4501 ctccctgccc attaaggaat cagagatcat tgatttcttc ctgggggcct ctctcaagga
     4561 tgaggttttg aagattatgc cagtgcagaa gcagacccgt gccggccagc gcaccaggtt
     4621 caaggcattt gttgctatcg gggactacaa tggccacgtc ggtctgggtg ttaagtgctc
     4681 caaggaggtg gccaccgcca tccgtggggc catcatcctg gccaagctct ccatcgtccc
     4741 cgtgcgcaga ggctactggg ggaacaagat cggcaagccc cacactgtcc cttgcaaggt
     4801 gacaggccgc tgcggctctg tgctggtacg cctcatccct gcacccaggg gcactggcat
     4861 cgtctccgca cctgtgccta agaagctgct catgatggct ggtatcgatg actgctacac
     4921 ctcagcccgg ggctgcactg ccaccctggg caacttcgcc aaggccacct ttgatgccat
     4981 ttctaagacc tacagctacc tgacccccga cctctggaag gagactgtat tcaccaagtc
     5041 tccctatcag gagttcactg accacctcgt caagacccac accagagtct ccgtgcagcg
     5101 gactcaggct ccagctgtgg ctacaacaaa cccagctttc ttgtacaaag tggttgcaag
     5161 ctttaaggcc ggtcctagca atatgagctc agagactggc ccagtggctg tggaccccac
     5221 attgagacgg cggatcgagc cccatgagtt tgaggtattc ttcgatccga gagagctccg
     5281 caaggagacc tgcctgcttt acgaaattaa ttgggggggc cggcactcca tttggcgaca
     5341 tacatcacag aacactaaca agcacgtcga agtcaacttc atcgagaagt tcacgacaga
     5401 aagatatttc tgtccgaaca caaggtgcag cattacctgg tttctcagct ggagcccatg
     5461 cggcgaatgt agtagggcca tcactgaatt cctgtcaagg tatccccacg tcactctgtt
     5521 tatttacatc gcaaggctgt accaccacgc tgacccccgc aatcgacaag gcctgcggga
     5581 tttgatctct tcaggtgtga ctatccaaat tatgactgag caggagtcag gatactgctg
     5641 gagaaacttt gtgaattata gcccgagtaa tgaagcccac tggcctaggt atccccatct
     5701 gtgggtacga ctgtacgttc ttgaactgta ctgcatcata ctgggcctgc ctccttgtct
     5761 caacattctg agaaggaagc agccacagct gacattcttt accatcgctc ttcagtcttg
     5821 tcattaccag cgactgcccc cacacattct ctgggccacc gggttgaaac tcgagtaccc
     5881 ctacgacgtg cccgactacg ccgctactaa cttcagcctg ctgaagcagg ctggagacgt
     5941 ggaggagaac cctggaccta gggtgtctaa gggcgaagag ctgatcaagg aaaatatgcg
     6001 tatgaaggtg gtcatggaag gttcggtcaa cggccaccaa ttcaaatgca caggtgaagg
     6061 agaaggcaga ccgtacgagg gagtgcaaac catgaggatc aaagtcatcg agggaggacc
     6121 cctgccattt gcctttgaca ttcttgccac gtcgttcatg tatggcagcc gtacctttat
     6181 caagtacccg gccgacatcc ctgatttctt taaacagtcc tttcctgagg gttttacttg
     6241 ggaaagagtt acgagatacg aagatggtgg agtcgtcacc gtcacgcagg acaccagcct
     6301 tgaggatggc gagctcgtct acaacgtcaa ggtcagaggg gtaaactttc cctccaatgg
     6361 tcccgtgatg cagaagaaga ccaagggttg ggagcctaat acagagatga tgtatccagc
     6421 agatggtggt ctgagaggat acactgacat cgcactgaaa gttgatggtg gtggccatct
     6481 gcactgcaac ttcgtgacaa cttacaggtc aaaaaagacc gtcgggaaca tcaagatgcc
     6541 cggtgtccat gccgttgatc accgcctgga aaggatcgag gagagtgaca atgaaaccta
     6601 cgtagtgcaa agagaagtgg cagttgccaa atacagcaac cttggtggtg gcatggacga
     6661 gctgtacaag taaaccggtc caccaccacc accaccacta aggatccggg gttggggttg
     6721 cgccttttcc aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc
     6781 gggaaacgca gcggcgccga ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc
     6841 acccggatct tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta
     6901 agtcgggaag gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt
     6961 ctcactagta ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga
     7021 tgggctgtgg ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg
     7081 gcggtgcggg aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt
     7141 tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc
     7201 accgacctct ctccccagca attcaccatg accgagtaca agcccacggt gcgcctcgcc
     7261 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     7321 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     7381 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     7441 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     7501 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     7561 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     7621 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     7681 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     7741 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     7801 tgcatgaccc gcaagcccgg tgccgaaggt agaggttctc tcctcacttg tggtgatgtt
     7861 gaagaaaacc ctggtccaat gtctagactg gacaagagca aagtcataaa cggagctctg
     7921 gaattactca atggtgtcgg tatcgaaggc ctgacgacaa ggaaactcgc tcaaaagctg
     7981 ggagttgagc agcctaccct gtactggcac gtgaagaaca agcgggccct gctcgatgcc
     8041 ctgccaatcg agatgctgga caggcatcat acccacttct gccccctgga aggcgagtca
     8101 tggcaagact ttctgcggaa caacgccaag tcataccgct gtgctctcct ctcacatcgc
     8161 gacggggcta aagtgcatct cggcacccgc ccaacagaga aacagtacga aaccctggaa
     8221 aatcagctcg cgttcctgtg tcagcaaggc ttctccctgg agaacgcact gtacgctctg
     8281 tccgccgtgg gccactttac actgggctgc gtattggagg aacaggagca tcaagtagca
     8341 aaagaggaaa gagagacacc taccaccgat tctatgcccc cacttctgag acaagcaatt
     8401 gagctgttcg accggcaggg agccgaacct gccttccttt tcggcctgga actaatcata
     8461 tgtggcctgg agaaacagct aaagtgcgaa agcggcgggc cgaccgacgc ccttgacgat
     8521 tttgacttag acatgctccc agccgatgcc cttgacgact ttgaccttga tatgctgcct
     8581 gctgacgctc ttgacgattt tgaccttgac atgctccccg ggtaactaag taaggatcga
     8641 tccaagatat cgtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     8701 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     8761 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     8821 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     8881 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     8941 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     9001 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     9061 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     9121 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     9181 cctttgggcc gcctccccgc ctgtttcgcc tcggcgtccg gactagaggt acctttaaga
     9241 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     9301 gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc
     9361 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     9421 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     9481 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt
     9541 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
     9601 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     9661 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     9721 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc
     9781 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     9841 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     9901 tttcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     9961 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
    10021 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
    10081 agttgggtgc acga
//
