LOCUS       sequence-374684-       10863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible lentiviral expression, TRE-gateway-APOBEC-HA-P2A-mRuby; 
            PGK-puro-2A-rtTA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10863)
  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 10863)
  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_Capture1_APOBEC_HA_P2A_mRuby
FEATURES             Location/Qualifiers
     source          1..10863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    41..158
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        212..526
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    220..238
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    256..274
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    291..309
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    327..345
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    363..381
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    398..416
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    434..452
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     499..523
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    547..671
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(696..701,702..719,720..726)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             780..1439
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(846..865)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1781..2086
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1965..1984
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2127..2251)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2280..2966
                     /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             2973..2999
                     /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             2973..2999
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3000..3056
                     /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             3060..3767
                     /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             3778..3795
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        3805..4315
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     4234..4252
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             4325..4921
                     /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(4325..4344)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4821..4841
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4922..4975
                     /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(4976..5593,5594..5599,5600..5635,5636..5638,5639..5674,
                     5675..5677,5678..5713,5714..5722)
                     /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(5762..5782)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6180..6191)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6396..6629
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6701..6822
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6738..6757)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6792..6811
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6862..6997
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6997..7068
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7069..7137,7138..7929)
                     /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(7287..7306)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8100..8688
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8589..8608
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8842..8859
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8976..8997
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9012..9017,9018..9035,9036..9042)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9050..9066
                     /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     9055..9077
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9074..9090
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9074..9090
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9109..9129
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9111..9129
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9157..9383
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             9384..9564
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9611..9736
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10229..10462
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10647..10691
                     /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"
ORIGIN
        1 aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
       61 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
      121 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
      181 gagatccact ttggccgcga atcgatatgt cgagtttact ccctatcagt gatagagaac
      241 gtatgtcgag tttactccct atcagtgata gagaacgatg tcgagtttac tccctatcag
      301 tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt
      361 actccctatc agtgatagag aacgtatgtc gagtttatcc ctatcagtga tagagaacgt
      421 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg
      481 ggaggcctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gaattggcta
      541 gcatcaacaa gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat
      601 atattaaatt agattttgca taaaaaacag actacataat actgtaaaac acaacatatc
      661 cagtcactat ggcggccgca ttaggcaccc caggctttac actttatgct tccggctcgt
      721 ataatgtgtg gattttgagt taggatccgt cgagattttc aggagctaag gaagctaaaa
      781 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
      841 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata
      901 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
      961 acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg aaagacggtg
     1021 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa
     1081 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt
     1141 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga
     1201 atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg
     1261 ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg
     1321 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg
     1381 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaaa
     1441 gatctggatc cggcttacta aaagccagat aacagtatgc gtatttgcgc gctgattttt
     1501 gcggtataag aatatatact gatatgtata cccgaagtat gtcaaaaaga ggtatgctat
     1561 gaagcagcgt attacagtga cagttgacag cgacagctat cagttgctca aggcatatat
     1621 gatgtcaata tctccggtct ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt
     1681 gccgaacgct ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg gtttattgaa
     1741 atgaacggct cttttgctga cgagaacagg ggctggtgaa atgcagttta aggtttacac
     1801 ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac
     1861 gcccgggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc
     1921 ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga
     1981 tatggccagt gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga
     2041 aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt caggctccct
     2101 tatacacagc cagtctgcag gtcgaccata gtgactggat atgttgtgtt ttacagtatt
     2161 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg
     2221 tttctcgttc agctttcttg tacaaagtgg ttgcaagctt taaggccggt cctagcaata
     2281 tgagctcaga gactggccca gtggctgtgg accccacatt gagacggcgg atcgagcccc
     2341 atgagtttga ggtattcttc gatccgagag agctccgcaa ggagacctgc ctgctttacg
     2401 aaattaattg ggggggccgg cactccattt ggcgacatac atcacagaac actaacaagc
     2461 acgtcgaagt caacttcatc gagaagttca cgacagaaag atatttctgt ccgaacacaa
     2521 ggtgcagcat tacctggttt ctcagctgga gcccatgcgg cgaatgtagt agggccatca
     2581 ctgaattcct gtcaaggtat ccccacgtca ctctgtttat ttacatcgca aggctgtacc
     2641 accacgctga cccccgcaat cgacaaggcc tgcgggattt gatctcttca ggtgtgacta
     2701 tccaaattat gactgagcag gagtcaggat actgctggag aaactttgtg aattatagcc
     2761 cgagtaatga agcccactgg cctaggtatc cccatctgtg ggtacgactg tacgttcttg
     2821 aactgtactg catcatactg ggcctgcctc cttgtctcaa cattctgaga aggaagcagc
     2881 cacagctgac attctttacc atcgctcttc agtcttgtca ttaccagcga ctgcccccac
     2941 acattctctg ggccaccggg ttgaaactcg agtaccccta cgacgtgccc gactacgccg
     3001 ctactaactt cagcctgctg aagcaggctg gagacgtgga ggagaaccct ggacctaggg
     3061 tgtctaaggg cgaagagctg atcaaggaaa atatgcgtat gaaggtggtc atggaaggtt
     3121 cggtcaacgg ccaccaattc aaatgcacag gtgaaggaga aggcagaccg tacgagggag
     3181 tgcaaaccat gaggatcaaa gtcatcgagg gaggacccct gccatttgcc tttgacattc
     3241 ttgccacgtc gttcatgtat ggcagccgta cctttatcaa gtacccggcc gacatccctg
     3301 atttctttaa acagtccttt cctgagggtt ttacttggga aagagttacg agatacgaag
     3361 atggtggagt cgtcaccgtc acgcaggaca ccagccttga ggatggcgag ctcgtctaca
     3421 acgtcaaggt cagaggggta aactttccct ccaatggtcc cgtgatgcag aagaagacca
     3481 agggttggga gcctaataca gagatgatgt atccagcaga tggtggtctg agaggataca
     3541 ctgacatcgc actgaaagtt gatggtggtg gccatctgca ctgcaacttc gtgacaactt
     3601 acaggtcaaa aaagaccgtc gggaacatca agatgcccgg tgtccatgcc gttgatcacc
     3661 gcctggaaag gatcgaggag agtgacaatg aaacctacgt agtgcaaaga gaagtggcag
     3721 ttgccaaata cagcaacctt ggtggtggca tggacgagct gtacaagtaa accggtccac
     3781 caccaccacc accactaagg atccggggtt ggggttgcgc cttttccaag gcagccctgg
     3841 gtttgcgcag ggacgcggct gctctgggcg tggttccggg aaacgcagcg gcgccgaccc
     3901 tgggtctcgc acattcttca cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct
     3961 tgtgggcccc ccggcgacgc ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt
     4021 cgcggcgtgc cggacgtgac aaacggaagc cgcacgtctc actagtaccc tcgcagacgg
     4081 acagcgccag ggagcaatgg cagcgcgccg accgcgatgg gctgtggcca atagcggctg
     4141 ctcagcaggg cgcgccgaga gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg
     4201 ggcggtagtg tgggccctgt tcctgcccgc gcggtgttcc gcattctgca agcctccgga
     4261 gcgcacgtcg gcagtcggct ccctcgttga ccgaatcacc gacctctctc cccagcaatt
     4321 caccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     4381 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     4441 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     4501 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     4561 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     4621 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     4681 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     4741 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     4801 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     4861 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     4921 cgaaggtaga ggttctctcc tcacttgtgg tgatgttgaa gaaaaccctg gtccaatgtc
     4981 tagactggac aagagcaaag tcataaacgg agctctggaa ttactcaatg gtgtcggtat
     5041 cgaaggcctg acgacaagga aactcgctca aaagctggga gttgagcagc ctaccctgta
     5101 ctggcacgtg aagaacaagc gggccctgct cgatgccctg ccaatcgaga tgctggacag
     5161 gcatcatacc cacttctgcc ccctggaagg cgagtcatgg caagactttc tgcggaacaa
     5221 cgccaagtca taccgctgtg ctctcctctc acatcgcgac ggggctaaag tgcatctcgg
     5281 cacccgccca acagagaaac agtacgaaac cctggaaaat cagctcgcgt tcctgtgtca
     5341 gcaaggcttc tccctggaga acgcactgta cgctctgtcc gccgtgggcc actttacact
     5401 gggctgcgta ttggaggaac aggagcatca agtagcaaaa gaggaaagag agacacctac
     5461 caccgattct atgcccccac ttctgagaca agcaattgag ctgttcgacc ggcagggagc
     5521 cgaacctgcc ttccttttcg gcctggaact aatcatatgt ggcctggaga aacagctaaa
     5581 gtgcgaaagc ggcgggccga ccgacgccct tgacgatttt gacttagaca tgctcccagc
     5641 cgatgccctt gacgactttg accttgatat gctgcctgct gacgctcttg acgattttga
     5701 ccttgacatg ctccccgggt aactaagtaa ggatcgatcc aagatatcgt attcttaact
     5761 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     5821 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     5881 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     5941 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     6001 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     6061 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat
     6121 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     6181 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     6241 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     6301 tttcgcctcg gcgtccggac tagaggtacc tttaagacca atgacttaca aggcagctgt
     6361 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     6421 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     6481 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     6541 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     6601 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     6661 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
     6721 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     6781 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     6841 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     6901 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     6961 ctattccaga agtagtgagg aggctttttt ggaggccttt caaatatgta tccgctcatg
     7021 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     7081 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     7141 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     7201 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     7261 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     7321 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     7381 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     7441 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     7501 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     7561 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     7621 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     7681 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     7741 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     7801 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     7861 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     7921 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     7981 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     8041 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     8101 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     8161 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     8221 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     8281 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     8341 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     8401 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     8461 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     8521 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     8581 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     8641 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     8701 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     8761 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     8821 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     8881 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     8941 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     9001 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     9061 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct
     9121 cactaaaggg aacaaaagct ggagctgcaa gcttaatgta gtcttatgca atactcttgt
     9181 agtcttgcaa catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc
     9241 gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga
     9301 cgggtctgac atggattgga cgaaccactg aattgccgca ttgcagagat attgtattta
     9361 agtgcctagc tcgatacata aacgggtctc tctggttaga ccagatctga gcctgggagc
     9421 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     9481 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     9541 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa
     9601 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga
     9661 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga
     9721 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg
     9781 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga
     9841 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat
     9901 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa
     9961 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc
    10021 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc
    10081 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
    10141 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
    10201 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
    10261 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
    10321 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
    10381 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
    10441 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
    10501 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
    10561 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
    10621 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
    10681 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
    10741 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga
    10801 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg
    10861 gac
//
