LOCUS       Exported               12403 bp DNA     circular SYN 04-JAN-2022
DEFINITION  Forward inducible lentivirus backbone coding for Delta variant 
            SARS-CoV-2 Spike (sinobiological optimization) with and 21aa CTD 
            deletion, and zsGreen that is linked to puromycin resistance gene by
            T2A linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12403)
  AUTHORS   Bernadeta Dadonaite
  TITLE     Direct Submission
  JOURNAL   Exported Jan 4, 2022 from SnapGene 6.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..12403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          5041..5058
                     /label=source
     misc_feature    9..186
                     /locus_tag="cppt"
                     /label=cppt
     misc_feature    33..150
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    163..164
                     /locus_tag="splicing_signal(1)"
                     /label=splicing_signal
                     /label=splicing_signal(1)
     misc_feature    188..192
                     /locus_tag="Insert from AAVS1"
                     /label=Insert from AAVS1
     misc_feature    188..192
                     /locus_tag="SpeI"
                     /label=SpeI
     misc_feature    197..202
                     /locus_tag="XhoI"
                     /label=XhoI
     misc_feature    200..567
                     /locus_tag="TRE3GS_promoter"
                     /label=TRE3GS_promoter
     promoter        200..567
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     misc_feature    208..226
                     /locus_tag="TetOperator"
                     /label=TetOperator
     protein_bind    208..226
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    244..262
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    280..298
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    316..334
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    352..370
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    388..406
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    424..442
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    587..606
                     /locus_tag="GAPDH_PrimingSite"
                     /label=GAPDH_PrimingSite
     primer_bind     591..634
                     /label=b11529_to_lenti_fw
     misc_feature    613..618
                     /locus_tag="KozakSeq"
                     /label=KozakSeq
     CDS             619..4368
                     /codon_start=1
                     /label=Spike
                     /translation="MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRS
                     SVLHSTQDLFLPFFSNVTWFHVISGTNGTKRFDNPVLPFNDGVYFASIEKSNIIRGWIF
                     GTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLDHKNNKSWMESEFRVYSSANNCTF
                     EYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPIIVREPEDLPQGFSALEP
                     LVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGT
                     ITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFDEVFN
                     ATRFASVYAWNRKRISNCVADYSVLYNLAPFFTFKCYGVSPTKLNDLCFTNVYADSFVI
                     RGDEVRQIAPGQTGNIADYNYKLPDDFTGCVIAWNSNKLDSKVSGNYNYLYRLFRKSNL
                     KPFERDISTEIYQAGNKPCNGVAGFNCYFPLRSYSFRPTYGVGHQPYRVVVLSFELLHA
                     PATVCGPKKSTNLVKNKCVNFNFNGLKGTGVLTESNKKFLPFQQFGRDIADTTDAVRDP
                     QTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQGVNCTEVPVAIHADQLTPTWRVYST
                     GSNVFQTRAGCLIGAEYVNNSYECDIPIGAGICASYQTQTKSHRRARSVASQSIIAYTM
                     SLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQY
                     GSFCTQLKRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKYFGGFNFSQILPDPSKPSKR
                     SFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFKGLTVLPPLLTDEMIAQYT
                     SALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGK
                     IQDSLSSTASALGKLQDVVNHNAQALNTLVKQLSSKFGAISSVLNDIFSRLDKVEAEVQ
                     IDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMS
                     FPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFY
                     EPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDI
                     SGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWYIWLGFIAGLIAIV
                     MVTIMLCCMTSCCSCLKGCCSCGS"
     misc_feature    817..819
                     /label=A67V
     misc_feature    822^823
                     /label=Delta-69-70
     misc_feature    895..897
                     /label=T95I
     primer_bind     complement(905..925)
                     
     misc_feature    1036..1038
                     /label=G142D
     misc_feature    1038^1039
                     /label=Delta-143-145
     misc_feature    1233^1234
                     /label=Delta-211
     misc_feature    1234..1236
                     /label=L212I
     misc_feature    1243..1251
                     /label=ins214EPE
     misc_feature    1624..1626
                     /label=G339D
     misc_feature    1720..1722
                     /label=S371L
     misc_feature    1726..1728
                     /label=S373P
     misc_feature    1732..1734
                     /label=S375F
     primer_bind     complement(1787..1812)
                     /label=8896_HDVSpA_Spike_SSF
     misc_feature    1858..1860
                     /label=K417N
     misc_feature    1927..1929
                     /label=N440K
     misc_feature    1945..1947
                     /label=G446S
     misc_feature    2038..2040
                     /label=S477N
     misc_feature    2041..2043
                     /label=T478K
     misc_feature    2059..2061
                     /label=E484A
     misc_feature    2086..2088
                     /label=Q493R
     misc_feature    2095..2097
                     /label=G496S
     misc_feature    2101..2103
                     /label=Q498R
     misc_feature    2110..2112
                     /label=N501Y
     misc_feature    2122..2124
                     /label=Y505H
     misc_feature    2248..2250
                     /label=T547K
     mat_peptide     2449..2451
                     /label=D614G
     misc_feature    2572..2574
                     /label=H655Y
     misc_feature    2644..2646
                     /label=N679K
     misc_feature    2650..2652
                     /label=P681H
     primer_bind     complement(2676..2699)
                     /label=9783_HDVSpA_Spike_SSF
     primer_bind     2729..2742
                     /label=b11529_to_lenti_rv
     misc_feature    2899..2901
                     /label=N764K
     misc_feature    2995..2997
                     /label=D796Y
     misc_feature    3175..3177
                     /label=N856K
     misc_feature    3469..3471
                     /label=Q954H
     misc_feature    3514..3516
                     /label=N969K
     primer_bind     complement(3537..3559)
                     /label=10644_HDVSpA_Spike_SSF
     misc_feature    3550..3552
                     /label=L981F
     primer_bind     complement(4346..4398)
                     /label=b11529_to_lenti_rv
     misc_feature    4375..4382
                     /locus_tag="AscI"
                     /label=AscI
     misc_feature    4393..4425
                     /locus_tag="IlluminaPrimerSite"
                     /label=IlluminaPrimerSite
     misc_feature    4426..4432
                     /locus_tag="XbaI"
                     /label=XbaI
     misc_feature    4432
                     /locus_tag="add_base_avoid_methylation"
                     /label=add_base_avoid_methylation
     promoter        4455..5039
                     /locus_tag="CMV IE promoter"
                     /label=CMV IE promoter
     enhancer        4464..4767
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4768..4971
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     TATA_signal     4959..4963
                     /locus_tag="CMV Promoter TATA Box"
                     /label=CMV Promoter TATA Box
     primer_bind     4983..5003
                     /locus_tag="primer_bind(1)"
                     /label=primer_bind(1)
     prim_transcript 4991
                     /locus_tag="CMV Transcriptional Start Site"
                     /label=CMV Transcriptional Start Site
     misc_feature    complement(5041..5058)
                     /label=T7 promoter
     regulatory      5061..5070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5067..5732
                     /locus_tag="ZsGreen Fluorescent Protein"
                     /label=ZsGreen Fluorescent Protein
     CDS             5733..5759
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="AIASGSALP"
     CDS             5778..5831
                     /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             5832..6431
                     /codon_start=1
                     /label=puroR
                     /note="Puromycin resistance gene"
                     /note="/vntifkey=4"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     6147..6168
                     /label=PuR_fw
     misc_feature    6416
                     /label=syn mut to add stop to frame 2
     misc_feature    6432..6449
                     /label=Feature 93
     misc_feature    6438..7029
                     /locus_tag="WPRE"
                     /label=WPRE
     misc_feature    6439..7027
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6466..6500)
                     /locus_tag="primer_bind"
                     /label=primer_bind
     primer_bind     complement(6466..6500)
                     /label=WPRE_PuR_rv
     misc_feature    7034..7100
                     /locus_tag="HIV nef"
                     /label=HIV nef
     misc_feature    7084..7098
                     /locus_tag="PolyPurine Tract"
                     /label=PolyPurine Tract
     LTR             7101..7736
                     /locus_tag="LTR(1)"
                     /label=LTR(1)
     LTR             7101..7734
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    7101..7554
                     /locus_tag="Full Length HIV U3(1)"
                     /label=Full Length HIV U3
                     /label=Full Length HIV U3(1)
     misc_feature    7555..7651
                     /locus_tag="HIV Repeat Region(1)"
                     /label=HIV Repeat Region
                     /label=HIV Repeat Region(1)
     polyA_site      7650
                     /locus_tag="Exact base to which Poly-A is added"
                     /label=Exact base to which Poly-A is added
     5'UTR           7651..7735
                     /locus_tag="HIV 5' UTR"
                     /label=HIV 5' UTR
     misc_feature    7652..7736
                     /locus_tag="HIV U5"
                     /label=HIV U5
     polyA_signal    7670..7677
                     /locus_tag="GT Cluster shown to be necessary 
                     forPoly-Asitea"
                     /label=GT Cluster shown to be necessary forPoly-A
                     site a
                     /label=GT Cluster shown to be necessary forPoly-A site a
     promoter        7869..7973
                     /gene="bla"
                     /label=AmpR promoter
     promoter        7939..7973
                     /locus_tag="Beta Lactamase Promoter"
                     /label=Beta Lactamase Promoter
     CDS             7974..8834
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             7992..8831
                     /locus_tag="beta-lactamase (mature form)"
                     /label=beta-lactamase (mature form)
     rep_origin      9005..9593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      9060..9593
                     /locus_tag="ColE1 origin of replication"
                     /label=ColE1 origin of replication
     misc_feature    9830..9953
                     /locus_tag="SV40 ori"
                     /label=SV40 ori
     protein_bind    10004..10025
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        10040..10070
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10078..10094
                     /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     10102..10118
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             10379..11014
                     /locus_tag="LTR"
                     /label=LTR
     LTR             10379..11012
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10379..10832
                     /locus_tag="Full Length HIV U3"
                     /label=Full Length HIV U3
     misc_feature    10833..10929
                     /locus_tag="HIV Repeat Region"
                     /label=HIV Repeat Region
     polyA_site      10928
                     /locus_tag="Exact base to which Poly-A is added(1)"
                     /label=Exact base to which Poly-A is added
                     /label=Exact base to which Poly-A is added(1)
     5'UTR           10929..11013
                     /locus_tag="HIV 5' UTR(1)"
                     /label=HIV 5' UTR
                     /label=HIV 5' UTR(1)
     misc_feature    10930..11014
                     /locus_tag="HIV U5(1)"
                     /label=HIV U5
                     /label=HIV U5(1)
     polyA_signal    10948..10955
                     /locus_tag="GT Cluster shown to be necessary 
                     forPoly-Asitea(1)"
                     /label=GT Cluster shown to be necessary forPoly-A
                     site a
                     /label=GT Cluster shown to be necessary forPoly-A sitea(1)
     misc_feature    11015..11533
                     /locus_tag="pbs-gag"
                     /label=pbs-gag
     misc_feature    11015..11167
                     /locus_tag="noncoding region upstream of Gag"
                     /label=noncoding region upstream of Gag
     variation       11032
                     /locus_tag="C->T relative to HIV pSF256.8"
                     /label=C->T relative to HIV pSF256.8
     misc_feature    11059..11184
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11075..11184
                     /locus_tag="misc_feature"
                     /label=misc_feature
     stem_loop       11075..11109
                     /locus_tag="PSI Stem loop 1"
                     /label=PSI Stem loop 1
     stem_loop       11114..11132
                     /locus_tag="PSI Stem loop 2"
                     /label=PSI Stem loop 2
     misc_feature    11122..11123
                     /locus_tag="Major 5' Splice Donor 289"
                     /label=Major 5' Splice Donor 289
                     /label=nonstandard type: splicing_signal
     stem_loop       11144..11157
                     /locus_tag="PSI Stem Loop 3"
                     /label=PSI Stem Loop 3
     CDS             11168..11522
                     /locus_tag="HIV gag sequence from 336bp to 
                     688bp(ClonetoPvu"
                     /label=HIV gag sequence from 336bp to 688bp(Clone
                     to Pvu
                     /label=HIV gag sequence from 336bp to 688bp(Clone to Pvu
     stem_loop       11171..11184
                     /locus_tag="PSI Stem Loop 4"
                     /label=PSI Stem Loop 4
     misc_feature    11185..11522
                     /locus_tag="HIV gag sequence 351bp to 688bp"
                     /label=HIV gag sequence 351bp to 688bp
     variation       11211..11212
                     /locus_tag="Filled in Cla Site from original 
                     HIVgenometodis"
                     /label=Filled in Cla Site from original HIVgenome
                     to dis
                     /label=Filled in Cla Site from original HIVgenome to dis
                     /label=nonstandard type: mutation
     misc_feature    11231..11233
                     /locus_tag="Stop codon in gag put in frame by ClaIfill in"
                     /label=Stop codon in gag put in frame by ClaIfill
                     in
                     /label=Stop codon in gag put in frame by ClaIfill in
     CDS             11534..12391
                     /locus_tag="HIV env sequence from bp7168 
                     to8025(containsRRE"
                     /label=HIV env sequence from bp7168 to
                     8025(contains RRE
                     /label=HIV env sequence from bp7168 to 8025(contains RRE
     misc_feature    11623..11971
                     /locus_tag="RRE"
                     /label=RRE
     misc_feature    11681..11914
                     /locus_tag="minimal RRE"
                     /label=minimal RRE
     variation       11735
                     /locus_tag="This G is a C in the WT HIV-1 genome"
                     /label=This G is a C in the WT HIV-1 genome
                     /label=nonstandard type: mutation
     misc_feature    12289..12290
                     /locus_tag="splicing_signal"
                     /label=splicing_signal
                     /label=nonstandard type: splicing_signal
     misc_feature    join(12392..12403,1..2)
                     /locus_tag="Linker Junk Sequence probably 
                     leftoverfromcloni"
                     /label=Linker Junk Sequence probably left
                     overfrom cloni
                     /label=Linker Junk Sequence probably left overfrom cloni
ORIGIN
        1 ccgaattcac aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
       61 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
      121 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
      181 gtttggacta gtctctctcg agtttactcc ctatcagtga tagagaacgt atgaagagtt
      241 tactccctat cagtgataga gaacgtatgc agactttact ccctatcagt gatagagaac
      301 gtataaggag tttactccct atcagtgata gagaacgtat gaccagttta ctccctatca
      361 gtgatagaga acgtatctac agtttactcc ctatcagtga tagagaacgt atatccagtt
      421 tactccctat cagtgataga gaacgtataa gctttgctta tgtaaaccag ggcgcctata
      481 aaagagtgct gattttttga gtaaacttca attccacaac acttttgtct tataccaact
      541 ttccgtacca cttcctaccc tcgtaaagaa ttcgcggccg cacgcgcagc cgagccacat
      601 cgctcaacgc gtgccaccat gtttgtgttc ctggtgctgc tgccactggt gtccagccag
      661 tgtgtgaacc tgaccaccag gacccaactt cctcctgcct acaccaactc cttcaccagg
      721 ggagtctact accctgacaa ggtgttcagg tcctctgtgc tgcacagcac ccaggacctg
      781 ttcctgccat tcttcagcaa tgtgacctgg ttccatgtga tctctggcac caatggcacc
      841 aagaggtttg acaaccctgt gctgccattc aatgatggag tctactttgc cagcatcgag
      901 aagagcaaca tcatcagggg ctggattttt ggcaccaccc tggacagcaa gacccagtcc
      961 ctgctgattg tgaacaatgc caccaatgtg gtgattaagg tgtgtgagtt ccagttctgt
     1021 aatgacccat tcctggacca caagaacaac aagtcctgga tggagtctga gttcagggtc
     1081 tactcctctg ccaacaactg tacctttgaa tatgtgagcc aaccattcct gatggacttg
     1141 gagggcaagc agggcaactt caagaacctg agggagtttg tgttcaagaa cattgatggc
     1201 tacttcaaga tttacagcaa acacacacca atcatcgtga gggagcccga ggacctgcca
     1261 cagggcttct ctgccttgga accactggtg gacctgccaa ttggcatcaa catcaccagg
     1321 ttccagaccc tgctggctct gcacaggtcc tacctgacac ctggagactc ctcctctggc
     1381 tggacagcag gagcagcagc ctactatgtg ggctacctcc aaccaaggac cttcctgctg
     1441 aaatacaatg agaatggcac catcacagat gctgtggact gtgccctgga cccactgtct
     1501 gagaccaagt gtaccctgaa atccttcaca gtggagaagg gcatctacca gaccagcaac
     1561 ttcagggtcc aaccaacaga gagcattgtg aggtttccaa acatcaccaa cctgtgtcca
     1621 tttgacgagg tgttcaatgc caccaggttt gcctctgtct atgcctggaa caggaagagg
     1681 attagcaact gtgtggctga ctactctgtg ctctacaacc tggccccctt cttcaccttc
     1741 aagtgttatg gagtgagccc aaccaaactg aatgacctgt gtttcaccaa tgtctatgct
     1801 gactcctttg tgattagggg agatgaggtg agacagattg cccctggaca aacaggcaac
     1861 attgctgact acaactacaa actgcctgat gacttcacag gctgtgtgat tgcctggaac
     1921 agcaacaagc tggacagcaa ggtgagcggc aactacaact acctctacag actgttcagg
     1981 aagagcaacc tgaaaccatt tgagagggac atcagcacag agatttacca ggctggcaac
     2041 aagccatgta atggagtggc cggcttcaac tgttactttc cactccgctc ctatagcttc
     2101 agaccaacct acggagtggg ccaccaacca tacagggtgg tggtgctgtc ctttgaactg
     2161 ctccatgccc ctgccacagt gtgtggacca aagaagagca ccaacctggt gaagaacaag
     2221 tgtgtgaact tcaacttcaa tggactgaag ggcacaggag tgctgacaga gagcaacaag
     2281 aagttcctgc cattccaaca gtttggcagg gacattgctg acaccacaga tgctgtgagg
     2341 gacccacaga ccttggagat tctggacatc acaccatgtt cctttggagg agtgtctgtg
     2401 attacacctg gcaccaacac cagcaaccag gtggctgtgc tctaccaggg tgtgaactgt
     2461 actgaggtgc ctgtggctat ccatgctgac caacttacac caacctggag ggtctacagc
     2521 acaggcagca atgtgttcca gaccagggct ggctgtctga ttggagcaga gtacgtgaac
     2581 aactcctatg agtgtgacat cccaattgga gcaggcatct gtgcctccta ccagacccag
     2641 accaagagcc acaggagggc aaggtctgtg gcaagccaga gcatcattgc ctacacaatg
     2701 agtctgggag cagagaactc tgtggcttac agcaacaaca gcattgccat cccaaccaac
     2761 ttcaccatct ctgtgaccac agagattctg cctgtgagta tgaccaagac ctctgtggac
     2821 tgtacaatgt atatctgtgg agacagcaca gagtgtagca acctgctgct ccaatatggc
     2881 tccttctgta cccaacttaa gagggctctg acaggcattg ctgtggaaca ggacaagaac
     2941 acccaggagg tgtttgccca ggtgaagcag atttacaaga cacctccaat caagtacttt
     3001 ggaggcttca acttcagcca gattctgcct gacccaagca agccaagcaa gaggtccttc
     3061 attgaggacc tgctgttcaa caaggtgacc ctggctgatg ctggcttcat caagcaatat
     3121 ggagactgtc tgggagacat tgctgccagg gacctgattt gtgcccagaa gttcaaggga
     3181 ctgacagtgc tgcctccact gctgacagat gagatgattg cccaatacac ctctgccctg
     3241 ctggctggca ccatcacctc tggctggacc tttggagcag gagcagccct ccaaatccca
     3301 tttgctatgc agatggctta caggttcaat ggcattggag tgacccagaa tgtgctctat
     3361 gagaaccaga aactgattgc caaccagttc aactctgcca ttggcaagat tcaggactcc
     3421 ctgtccagca cagcctctgc cctgggcaaa ctccaagatg tggtgaacca caatgcccag
     3481 gctctgaaca ccctggtgaa gcaactttcc agcaagtttg gagccatctc ctctgtgctg
     3541 aatgacatct tcagcagact ggacaaggtg gaggctgagg tccagattga cagactgatt
     3601 acaggcagac tccaatccct ccaaacctat gtgacccaac aacttatcag ggctgctgag
     3661 attagggcat ctgccaacct ggctgccacc aagatgagtg agtgtgtgct gggacaaagc
     3721 aagagggtgg acttctgtgg caagggctac cacctgatga gttttccaca gtctgcccct
     3781 catggagtgg tgttcctgca tgtgacctat gtgcctgccc aggagaagaa cttcaccaca
     3841 gcccctgcca tctgccatga tggcaaggct cactttccaa gggagggagt gtttgtgagc
     3901 aatggcaccc actggtttgt gacccagagg aacttctatg aaccacagat tatcaccaca
     3961 gacaacacct ttgtgtctgg caactgtgat gtggtgattg gcattgtgaa caacacagtc
     4021 tatgacccac tccaacctga actggactcc ttcaaggagg aactggacaa atacttcaag
     4081 aaccacacca gccctgatgt ggacctggga gacatctctg gcatcaatgc ctctgtggtg
     4141 aacatccaga aggagattga cagactgaat gaggtggcta agaacctgaa tgagtccctg
     4201 attgacctcc aagaactggg caaatatgaa caatacatca agtggccatg gtacatctgg
     4261 ctgggcttca ttgctggact gattgcaatt gtgatggtga caataatgct gtgttgtatg
     4321 acctcctgtt gttcctgtct gaaaggctgt tgttcctgtg gctcctgata atgaggcgcg
     4381 cctagttgga tcacactctt tccctacacg acgctcttcc gatcttctag attcgagaga
     4441 gaaatgttcc tagtggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     4501 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     4561 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     4621 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     4681 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     4741 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     4801 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     4861 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     4921 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
     4981 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
     5041 ctatagtgag tcgtattacg gccgccatgg cccagtccaa gcacggcctg accaaggaga
     5101 tgaccatgaa gtaccgcatg gagggctgcg tggacggcca caagttcgtg atcaccggcg
     5161 agggcatcgg ctaccccttc aagggcaagc aggccatcaa cctgtgcgtg gtggagggcg
     5221 gccccttgcc cttcgccgag gacatcttgt ccgccgcctt catgtacggc aaccgcgtgt
     5281 tcaccgagta cccccaggac atcgtcgact acttcaagaa ctcctgcccc gccggctaca
     5341 cctgggaccg ctccttcctg ttcgaggacg gcgccgtgtg catctgcaac gccgacatca
     5401 ccgtgagcgt ggaggagaac tgcatgtacc acgagtccaa gttctacggc gtgaacttcc
     5461 ccgccgacgg ccccgtgatg aagaagatga ccgacaactg ggagccctcc tgcgagaaga
     5521 tcatccccgt gcccaagcag ggcatcttga agggcgacgt gagcatgtac ctgctgctga
     5581 aggacggtgg ccgcttgcgc tgccagttcg acaccgtgta caaggccaag tccgtgcccc
     5641 gcaagatgcc cgactggcac ttcatccagc acaagctgac ccgcgaggac cgcagcgacg
     5701 ccaagaacca gaagtggcac ctgaccgagc acgccatcgc ctccggctcc gccttgcccg
     5761 ctggatccgg ctccggcgag ggcaggggaa gtcttctaac atgcggggac gtggaggaaa
     5821 atcccggccc aatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     5881 cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     5941 tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     6001 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     6061 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     6121 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     6181 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     6241 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     6301 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     6361 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgtaagc
     6421 ccggtgcctg aggatcctaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     6481 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     6541 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     6601 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     6661 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     6721 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     6781 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     6841 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     6901 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     6961 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     7021 tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     7081 tttaaaagaa aaggggggac tggaagggct aattcactcc caaagaagac aagatatcct
     7141 tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg
     7201 gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc
     7261 agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct
     7321 gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc
     7381 atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc
     7441 ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg
     7501 ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc
     7561 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     7621 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     7681 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtcccg
     7741 ggcgattaag gaaagggcta gatcattctt gaagacgaaa gggcctcgtg atacgcctat
     7801 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     7861 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     7921 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     7981 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     8041 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     8101 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     8161 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     8221 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     8281 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     8341 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     8401 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     8461 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     8521 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     8581 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     8641 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     8701 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     8761 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     8821 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     8881 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     8941 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     9001 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     9061 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     9121 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     9181 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     9241 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     9301 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     9361 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     9421 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     9481 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     9541 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     9601 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     9661 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     9721 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     9781 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc aagctcatgg
     9841 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     9901 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc ccgtggcacg
     9961 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
    10021 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
    10081 tgagcggata acaatttcac acaggaaaca gctatgacat gattacgaat ttcacaaata
    10141 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
    10201 atgtctggat caactggata actcaagcta accaaaatca tcccaaactt cccaccccat
    10261 accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg
    10321 aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagttg
    10381 gaagggctaa ttcactccca aagaagacaa gatatccttg atctgtggat ctaccacaca
    10441 caaggctact tccctgatta gcagaactac acaccagggc caggggtcag atatccactg
    10501 acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga agaggccaat
    10561 aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga tgacccggag
    10621 agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt ggcccgagag
    10681 ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg actttccgct
    10741 ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc cctcagatcc
    10801 tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc
    10861 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
    10921 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
    10981 acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg
    11041 aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca
    11101 agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag
    11161 gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa
    11221 aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca
    11281 agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt
    11341 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca
    11401 ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc
    11461 aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa
    11521 gcggccggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
    11581 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
    11641 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
    11701 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
    11761 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
    11821 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
    11881 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
    11941 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
    12001 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
    12061 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
    12121 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
    12181 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
    12241 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
    12301 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
    12361 gagacagatc cattcgatta gtgaacggat ctcgacggta tcg
//
