LOCUS       sequence-365343-        9152 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible lentiviral expression, TRE-control-APOBEC-HA-P2A-mRuby; 
            PGK-puro-2A-rtTA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9152)
  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 9152)
  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_control_APOBEC_HA_P2A_mRuby_Capture1
FEATURES             Location/Qualifiers
     source          1..9152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6..692
                     /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             699..725
                     /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             699..725
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             726..782
                     /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             786..1493
                     /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"
     promoter        1521..2031
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1950..1968
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2041..2637
                     /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(2041..2060)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2537..2557
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2638..2691
                     /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(2692..3309,3310..3315,3316..3351,3352..3354,3355..3390,
                     3391..3393,3394..3429,3430..3438)
                     /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(3478..3498)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3896..3907)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4112..4345
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4417..4538
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4454..4473)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4508..4527
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4578..4713
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4713..4784
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4785..4853,4854..5645)
                     /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(5003..5022)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5816..6404
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6305..6324
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6558..6575
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6692..6713
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6728..6733,6734..6751,6752..6758)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6766..6782
                     /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     6771..6793
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6790..6806
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6790..6806
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6825..6845
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6827..6845
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6873..7099
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7100..7280
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7327..7452
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7945..8178
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8363..8407
                     /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    8620..8737
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        8791..9105
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    8799..8817
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8835..8853
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8870..8888
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8906..8924
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8942..8960
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8977..8995
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9013..9031
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     9078..9102
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    9126..9150
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 cgcgtatgag ctcagagact ggcccagtgg ctgtggaccc cacattgaga cggcggatcg
       61 agccccatga gtttgaggta ttcttcgatc cgagagagct ccgcaaggag acctgcctgc
      121 tttacgaaat taattggggg ggccggcact ccatttggcg acatacatca cagaacacta
      181 acaagcacgt cgaagtcaac ttcatcgaga agttcacgac agaaagatat ttctgtccga
      241 acacaaggtg cagcattacc tggtttctca gctggagccc atgcggcgaa tgtagtaggg
      301 ccatcactga attcctgtca aggtatcccc acgtcactct gtttatttac atcgcaaggc
      361 tgtaccacca cgctgacccc cgcaatcgac aaggcctgcg ggatttgatc tcttcaggtg
      421 tgactatcca aattatgact gagcaggagt caggatactg ctggagaaac tttgtgaatt
      481 atagcccgag taatgaagcc cactggccta ggtatcccca tctgtgggta cgactgtacg
      541 ttcttgaact gtactgcatc atactgggcc tgcctccttg tctcaacatt ctgagaagga
      601 agcagccaca gctgacattc tttaccatcg ctcttcagtc ttgtcattac cagcgactgc
      661 ccccacacat tctctgggcc accgggttga aactcgagta cccctacgac gtgcccgact
      721 acgccgctac taacttcagc ctgctgaagc aggctggaga cgtggaggag aaccctggac
      781 ctagggtgtc taagggcgaa gagctgatca aggaaaatat gcgtatgaag gtggtcatgg
      841 aaggttcggt caacggccac caattcaaat gcacaggtga aggagaaggc agaccgtacg
      901 agggagtgca aaccatgagg atcaaagtca tcgagggagg acccctgcca tttgcctttg
      961 acattcttgc cacgtcgttc atgtatggca gccgtacctt tatcaagtac ccggccgaca
     1021 tccctgattt ctttaaacag tcctttcctg agggttttac ttgggaaaga gttacgagat
     1081 acgaagatgg tggagtcgtc accgtcacgc aggacaccag ccttgaggat ggcgagctcg
     1141 tctacaacgt caaggtcaga ggggtaaact ttccctccaa tggtcccgtg atgcagaaga
     1201 agaccaaggg ttgggagcct aatacagaga tgatgtatcc agcagatggt ggtctgagag
     1261 gatacactga catcgcactg aaagttgatg gtggtggcca tctgcactgc aacttcgtga
     1321 caacttacag gtcaaaaaag accgtcggga acatcaagat gcccggtgtc catgccgttg
     1381 atcaccgcct ggaaaggatc gaggagagtg acaatgaaac ctacgtagtg caaagagaag
     1441 tggcagttgc caaatacagc aaccttggtg gtggcatgga cgagctgtac aagtaaagct
     1501 ttaaggccgg tcctagcaac ggggttgggg ttgcgccttt tccaaggcag ccctgggttt
     1561 gcgcagggac gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg
     1621 tctcgcacat tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg
     1681 ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg
     1741 gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc agacggacag
     1801 cgccagggag caatggcagc gcgccgaccg cgatgggctg tggccaatag cggctgctca
     1861 gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg
     1921 gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc
     1981 acgtcggcag tcggctccct cgttgaccga atcaccgacc tctctcccca gcaattcacc
     2041 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     2101 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     2161 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     2221 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     2281 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     2341 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     2401 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     2461 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     2521 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     2581 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgccgaa
     2641 ggtagaggtt ctctcctcac ttgtggtgat gttgaagaaa accctggtcc aatgtctaga
     2701 ctggacaaga gcaaagtcat aaacggagct ctggaattac tcaatggtgt cggtatcgaa
     2761 ggcctgacga caaggaaact cgctcaaaag ctgggagttg agcagcctac cctgtactgg
     2821 cacgtgaaga acaagcgggc cctgctcgat gccctgccaa tcgagatgct ggacaggcat
     2881 catacccact tctgccccct ggaaggcgag tcatggcaag actttctgcg gaacaacgcc
     2941 aagtcatacc gctgtgctct cctctcacat cgcgacgggg ctaaagtgca tctcggcacc
     3001 cgcccaacag agaaacagta cgaaaccctg gaaaatcagc tcgcgttcct gtgtcagcaa
     3061 ggcttctccc tggagaacgc actgtacgct ctgtccgccg tgggccactt tacactgggc
     3121 tgcgtattgg aggaacagga gcatcaagta gcaaaagagg aaagagagac acctaccacc
     3181 gattctatgc ccccacttct gagacaagca attgagctgt tcgaccggca gggagccgaa
     3241 cctgccttcc ttttcggcct ggaactaatc atatgtggcc tggagaaaca gctaaagtgc
     3301 gaaagcggcg ggccgaccga cgcccttgac gattttgact tagacatgct cccagccgat
     3361 gcccttgacg actttgacct tgatatgctg cctgctgacg ctcttgacga ttttgacctt
     3421 gacatgctcc ccgggtaact aagtaaggat cgatccaaga tatcgtattc ttaactatgt
     3481 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3541 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     3601 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3661 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     3721 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3781 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     3841 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     3901 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3961 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctgtttc
     4021 gcctcggcgt ccggactaga ggtaccttta agaccaatga cttacaaggc agctgtagat
     4081 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     4141 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     4201 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     4261 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     4321 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     4381 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     4441 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4501 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     4561 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4621 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4681 tccagaagta gtgaggaggc ttttttggag gcctttcaaa tatgtatccg ctcatgagac
     4741 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4801 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4861 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4921 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4981 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5041 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5101 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5161 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5221 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5281 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5341 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5401 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5461 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5521 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5581 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5641 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5701 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5761 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5821 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5881 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5941 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6001 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6061 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6121 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6181 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6241 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6301 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6361 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6421 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6481 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6541 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     6601 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     6661 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     6721 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     6781 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     6841 aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc
     6901 ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc
     6961 atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg
     7021 tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg
     7081 cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc
     7141 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     7201 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     7261 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
     7321 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
     7381 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
     7441 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
     7501 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     7561 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     7621 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     7681 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
     7741 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat
     7801 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     7861 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     7921 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     7981 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     8041 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     8101 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     8161 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     8221 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     8281 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     8341 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     8401 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     8461 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     8521 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggaca
     8581 attctcgacc tcgagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     8641 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     8701 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     8761 agatccactt tggccgcgaa tcgatatgtc gagtttactc cctatcagtg atagagaacg
     8821 tatgtcgagt ttactcccta tcagtgatag agaacgatgt cgagtttact ccctatcagt
     8881 gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgagttta
     8941 ctccctatca gtgatagaga acgtatgtcg agtttatccc tatcagtgat agagaacgta
     9001 tgtcgagttt actccctatc agtgatagag aacgtatgtc gaggtaggcg tgtacggtgg
     9061 gaggcctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag aattggctag
     9121 catcaacaag tttgtacaaa aaagcaggct ta
//
