LOCUS       Exported                9995 bp ds-DNA     circular SYN 03-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9995)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 5, 2023 from SnapGene 4.0.8
            http://www.snapgene.com
COMMENT     Notes o The plasmid has been transformed in SURE2 cells 
            (Stratagene), to avoid recombination. o The cassette 
            [5'TR-MCS-pA-3'TR] has been cloned between the SacII and KpnI sites 
            of the vector pBluescript II SK+. It can be removed by cutting 
            SacII-KpnI or by cutting with AscI. o The 5'TR contains the two 
            point mutations described in Lacoste et al., 2009 (Cell Stem Cell). 
            o The following primers can be used to sequence the plasmid: M13-rev
            and T3 (upstream of the 5'TR, top strand); M13-fw and T7 (downstream
            of the 3'TR, bottom strand). o Any restriction site can be used to 
            clone your favorite promoter/gene. However, I would suggest cloning 
            the promoter between XhoI-BamHI and the gene between BamHI-NotI, for
            consistency with previous constructs by Arnaud and others in the 
            lab. o You don't need to add a polyAdenylation (pA) sequence, since 
            it is already present in the vector. If undesired, the pA can be 
            removed with the EcoRI site.
FEATURES             Location/Qualifiers
     source          1..9995
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          326..557
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     source          3219..3281
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     source          6189..6419
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     repeat_region   1..325
                     /label=5' PB Terminal Repeat (ePB)
                     /note="5' Piggybac Transposon Terminal Repeat. Optimized 
                     for human IPSC by Arnaud et al. 2009."
     misc_feature    14..26
                     /label=13 bp terminal repeat (TR)
     misc_feature    27..29
                     /label=3 bp spacer
     misc_feature    30..48
                     /label=19 bp Internal Repeat (IR)
     enhancer        326..557
                     /label=core Insulator
                     /label=core insul enhancer
                     /label=core insul
     misc_feature    564..590
                     /label=I-CeuI site
                     /label=ICeuI site
                     /note="ICeuI site"
     misc_feature    591..605
                     /label=AarI site
     promoter        612..861
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA. 
                     Taken from Feng Zhang lab CRISPR sgRNA plasmids (Addgene 
                     42230, 60958, 61591)."
     primer_bind     612..632
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     783..802
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     regulatory      832..837
                     /regulatory_class="TATA_box"
     misc_signal     861
                     /label=TSS +1
                     /note="Transcription start site for human U6 promoter. 
                     First base of transcript. G is preferred."
     regulatory      881..886
                     /regulatory_class="terminator"
     misc_feature    887..901
                     /label=AarI site
     misc_feature    894..911
                     /label=ISceI site
                     /note="ISceI site"
     promoter        join(920..1175,1180..1313)
                     /label=TRE3GS promoter
     protein_bind    928..946
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    964..982
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1000..1018
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1036..1054
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1072..1090
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1108..1126
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1144..1162
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    1176..1179
                     /label=Overlap for assembly o2
     regulatory      1326..1335
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1332..2831
                     /codon_start=1
                     /product="Rev-independent version of HIV-1 Gag coding 
                     sequence from HXB2 [pCMV55M1-10 obtained from Dr. George 
                     Pavlakis, NCI (Schwartz et al (1992) J. Virol. 66: 
                     7176-7182]. "
                     /label=HIV-1 gag
                     /translation="MGARASVLSGGELDRWEKIRLRPGGKKKYKLKHIVWASRELERFA
                     VNPGLLETSEGCRQILGQLQPSLQTGSEELRSLYNTVATLYCVHQRIEIKDTKEALDKI
                     EEEQNKSKKKAQQAAADTGHSNQVSQNYPIVQNIQGQMVHQAISPRTLNAWVKVVEEKA
                     FSPEVIPMFSALSEGATPQDLNTMLNTVGGHQAAMQMLKETINEEAAEWDRVHPVHAGP
                     IAPGQMREPRGSDIAGTTSTLQEQIGWMTNNPPIPVGEIYKRWIILGLNKIVRMYSPTS
                     ILDIRQGPKEPFRDYVDRFYKTLRAEQASQEVKNWMTETLLVQNANPDCKTILKALGPA
                     ATLEEMMTACQGVGGPGHKARVLAEAMSQVTNSATIMMQRGNFRNQRKIVKCFNCGKEG
                     HTARNCRAPRKKGCWKCGKEGHQMKDCTERQANFLGKIWPSYKGRPGNFLQSRPEPTAP
                     PEESFRSGVETTTPPQKQEPIDKELYPLTSLRSLFGNDPSSQ"
     CDS             1335..1727
                     /product="HIV-1 Matrix protein p17 
                     (https://www.uniprot.org/uniprot/P04591)"
                     /label=HIV-1 MA
     CDS             1728..2420
                     /label=HIV-1 CA
     misc_feature    2247..2253
                     /label=Recoded to remove Bpu1102I site
                     /note="Synonymously recoded to remove Bpu1102I site 
                     (GCTGAGC -> GCGGAAC)"
     CDS             2421..2462
                     /product="HIV-1 Spacer peptide 1 
                     (https://www.uniprot.org/uniprot/P04591)"
                     /label=HIV-1 SP1
     CDS             2463..2627
                     /label=HIV-1 NC
     CDS             2499..2552
                     /product="Zinc finger domain 1 
                     (https://www.uniprot.org/uniprot/P04591), CCHC-type 1"
                     /label=ZF1
     CDS             2562..2615
                     /product="Zinc finger domain 2 
                     (https://www.uniprot.org/uniprot/P04591), CCHC-type 2"
                     /label=ZF2
     misc_feature    2625..2684
                     /label=Recoded to remove gag-pol frameshift
                     /note="Recoded synonymously using IDT Codon Optimizer for 
                     Human to remove gag-pol frameshift sequences."
     CDS             2628..2675
                     /product="HIV-1 Spacer peptide 2 
                     (https://www.uniprot.org/uniprot/P04591)"
                     /label=HIV-1 SP2
     CDS             2676..2684
                     /product="Plays a role in budding of the assembled particle
                     by interacting with the host class E VPS proteins TSG101 
                     and PDCD6IP/AIP1."
                     /label=HIV-1 p6
     CDS             2685..2831
                     /product="Plays a role in budding of the assembled particle
                     by interacting with the host class E VPS proteins TSG101 
                     and PDCD6IP/AIP1."
                     /label=HIV-1 p6
     CDS             2694..2705
                     /product="Late-budding domains (L domains) are short 
                     sequence motifs essential for viral particle budding. They 
                     recruit proteins of the host ESCRT machinery (Endosomal 
                     Sorting Complex Required for Transport) or ESCRT-associated
                     proteins. p6-gag contains two L domains: a PTAP/PSAP motif,
                     which interacts with the UEV domain of TSG101 and a 
                     LYPX(n)L motif which interacts with PDCD6IP/AIP1 
                     (https://www.uniprot.org/uniprot/P04591)"
                     /label=HIV-1 p6-gag L domain
     CDS             2778..2807
                     /product="Late-budding domains (L domains) are short 
                     sequence motifs essential for viral particle budding. They 
                     recruit proteins of the host ESCRT machinery (Endosomal 
                     Sorting Complex Required for Transport) or ESCRT-associated
                     proteins. p6-gag contains two L domains: a PTAP/PSAP motif,
                     which interacts with the UEV domain of TSG101 and a 
                     LYPX(n)L motif which interacts with PDCD6IP/AIP1 
                     (https://www.uniprot.org/uniprot/P04591)"
                     /label=HIV-1 p6-gag L domain
     CDS             2832..3218
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2-N55K
                     /note="binds with high affinity to a specific stem-loop 
                     structure in the viral RNA (Lim et al., 1994)"
     CDS             3219..3281
                     /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="GSGEGRGSLLTCGDVEENPGP"
     CDS             3282..3998
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     polyA_signal    4013..4249
                     /label=pA
                     /note="pA"
     polyA_signal    4126..4247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    4258..4707
                     /label=PGK promoter
                     /label=PGK(-)
                     /note="/ApEinfo_fwdcolor=#ff8000
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
                     /ApEinfo_revcolor=#ff8040"
     regulatory      4717..4725
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4723..5466
                     /codon_start=1
                     /label=Tet-On 3G
                     /label=Tet-On\3G
                     /note="/ApEinfo_fwdcolor=#0080ff
                     /ApEinfo_revcolor=#0080ff"
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             5467..5529
                     /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="GSGEGRGSLLTCGDVEENPGP"
     CDS             5530..5928
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BlastR
                     /note="BSD
                     confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5946..6170
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        complement(6189..6419)
                     /label=core Insulator
                     /label=core insul(1) enhancer
                     /label=core insul(1)
     repeat_region   6420..6672
                     /label=3' PB Terminal Repeat (ePB)
                     /note="3' Piggybac Transposon Terminal Repeat. Optimized 
                     for human IPSC by Arnaud et al. 2009."
     misc_feature    6597..6615
                     /label=19 bp Internal Repeat (IR)
     misc_feature    6616..6646
                     /label=31 bp spacer
     misc_feature    6647..6659
                     /label=13 bp terminal repeat (TR)
     primer_bind     complement(7158..7174)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7182..7198
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(7206..7236)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7251..7272
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7560..8148)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8319..9113)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(9114..9218)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(9244..9699)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     9841..9857
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gcagctagat taaccctaga aagatagtct gcgtaaaatt gacgcatgca ttcttgaaat
       61 attgctctct ctttctaaat agcgcgaatc cgtcgctgtg catttaggac atctcagtcg
      121 ccgcttggag ctcccgtgag gcgtgcttgt caatgcggta agtgtcactg attttgaact
      181 ataacaaccg cgtgagtcaa aatgacgcat gattatcttt tacgtgactt ttaagattta
      241 actcatacga taattatatt gttatttcgt gttctactta cgtgataact tattatatat
      301 atattttctt gttatagata tccttgaggg acagcccccc cccaaagccc ccagggatgt
      361 aattacgtcc ctcccccgct agggggcagc agcgagccgc ccggggctcc gctccggtcc
      421 ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga cagcccgggc acggggaagg
      481 tggcacggga tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg cagacacctg
      541 gggggatacg gggaaaaatc gattaactat aacggtccta aggtagcgaa cacctgcgct
      601 cgagcgacgt cgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
      661 ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata
      721 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
      781 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
      841 tgtggaaagg acgaaacacc gaccaggtgg gcccacgcgt ttttttcacc tgcattaccc
      901 tgttatccct acctgcaggg agtttactcc ctatcagtga tagagaacgt atgaagagtt
      961 tactccctat cagtgataga gaacgtatgc agactttact ccctatcagt gatagagaac
     1021 gtataaggag tttactccct atcagtgata gagaacgtat gaccagttta ctccctatca
     1081 gtgatagaga acgtatctac agtttactcc ctatcagtga tagagaacgt atatccagtt
     1141 tactccctat cagtgataga gaacgtataa gctttcgtat gatagagaac gtataagctt
     1201 tgcttatgta aaccagggcg cctataaaag agtgctgatt ttttgagtaa acttcaattc
     1261 cacaacactt ttgtcttata ccaactttcc gtaccacttc ctaccctcgt aaagctagcg
     1321 gatccgccac catgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgatggg
     1381 aaaaaattcg gttaaggcca gggggaaaga agaagtacaa gctaaagcac atcgtatggg
     1441 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
     1501 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagag gagcttcgat
     1561 cactatacaa cacagtagca accctctatt gtgtgcacca gcggatcgag atcaaggaca
     1621 ccaaggaagc tttagacaag atagaggaag agcaaaacaa gtccaagaag aaggcccagc
     1681 aggcagcagc tgacacagga cacagcaatc aggtcagcca aaattaccct atagtgcaga
     1741 acatccaggg gcaaatggta catcaggcca tatcacctag aactttaaat gcatgggtaa
     1801 aagtagtaga agagaaggct ttcagcccag aagtgatacc catgttttca gcattatcag
     1861 aaggagccac cccacaggac ctgaacacga tgttgaacac cgtgggggga catcaagcag
     1921 ccatgcaaat gttaaaagag accatcaatg aggaagctgc agaatgggat agagtgcatc
     1981 cagtgcatgc agggcctatt gcaccaggcc agatgagaga accaagggga agtgacatag
     2041 caggaactac tagtaccctt caggaacaaa taggatggat gacaaataat ccacctatcc
     2101 cagtaggaga gatctacaag aggtggataa tcctgggatt gaacaagatc gtgaggatgt
     2161 atagccctac cagcattctg gacataagac aaggaccaaa agaacccttt agagactatg
     2221 tagaccggtt ctataaaact ctaagagcgg aacaagcttc acaggaggta aaaaattgga
     2281 tgacagaaac cttgttggtc caaaatgcga acccagattg taagaccatc ctgaaggctc
     2341 tcggcccagc ggctacacta gaagaaatga tgacagcatg tcagggagta ggaggacccg
     2401 gccataaggc aagagttttg gctgaagcaa tgagccaagt aacaaattca gctaccataa
     2461 tgatgcagag aggcaatttt aggaaccaaa gaaagattgt taagtgtttc aattgtggca
     2521 aagaagggca cacagccaga aattgcaggg cccctaggaa aaagggctgt tggaaatgtg
     2581 gaaaggaagg acaccaaatg aaagattgta ctgagagaca ggctaacttt ctggggaaaa
     2641 tttggcccag ctacaaaggt aggccgggaa atttcctgca atcaagacca gagccaacag
     2701 ccccaccaga agagagcttc aggtctgggg tagagacaac aactccccct cagaagcagg
     2761 agccgataga caaggaactg tatcctttaa cttccctcag atcactcttt ggcaacgacc
     2821 cctcgtcaca agcttcaaac tttactcagt tcgtgctcgt ggacaatggt gggacagggg
     2881 atgtgacagt ggctccttct aatttcgcta atggggtggc agagtggatc agctccaact
     2941 cacggagcca ggcctacaag gtgacatgca gcgtcaggca gtctagtgcc cagaagagaa
     3001 agtataccat caaggtggag gtccccaaag tggctaccca gacagtgggc ggagtcgaac
     3061 tgcctgtcgc cgcttggagg tcctacctga acatggagct cactatccca attttcgcta
     3121 ccaattctga ctgtgaactc atcgtgaagg caatgcaggg gctcctcaaa gacggtaatc
     3181 ctatcccttc cgccatcgcc gctaactcag gtatctacgg atcgggagag ggcagaggaa
     3241 gtctgctaac atgcggtgac gtcgaggaga atcctggccc aatgagcaag ggcgaggagc
     3301 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     3361 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     3421 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     3481 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     3541 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     3601 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     3661 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     3721 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     3781 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     3841 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     3901 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     3961 ccgggatcac tcacggcatg gacgagctgt acaagtaagc ggccgcgtcg acgactctag
     4021 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     4081 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     4141 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4201 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagc gccggcgcta
     4261 cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc
     4321 gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc
     4381 cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct
     4441 cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca
     4501 atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg
     4561 cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc
     4621 attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg
     4681 cctttcgacc tgcagcccaa gcttaccatg ggtaccgcca ccatgtctag actggacaag
     4741 agcaaagtca taaactctgc tctggaatta ctcaatggag tcggtatcga aggcctgacg
     4801 acaaggaaac tcgctcaaaa gctgggagtt gagcagccta ccctgtactg gcacgtgaag
     4861 aacaagcggg ccctgctcga tgccctgcca atcgagatgc tggacaggca tcatacccac
     4921 tcctgccccc tggaaggcga gtcatggcaa gactttctgc ggaacaacgc caagtcatac
     4981 cgctgtgctc tcctctcaca tcgcgacggg gctaaagtgc atctcggcac ccgcccaaca
     5041 gagaaacagt acgaaaccct ggaaaatcag ctcgcgttcc tgtgtcagca aggcttctcc
     5101 ctggagaacg cactgtacgc tctgtccgcc gtgggccact ttacactggg ctgcgtattg
     5161 gaggaacagg agcatcaagt agcaaaagag gaaagagaga cacctaccac cgattctatg
     5221 cccccacttc tgaaacaagc aattgagctg ttcgaccggc agggagccga acctgccttc
     5281 cttttcggcc tggaactaat catatgtggc ctggagaaac agctaaagtg cgaaagcggc
     5341 gggccgaccg acgcccttga cgattttgac ttagacatgc tcccagccga tgcccttgac
     5401 gactttgacc ttgatatgct gcctgctgac gctcttgacg attttgacct tgacatgctc
     5461 cccgggggat ctggtgaggg gcggggaagt ttgcttacgt gcggagatgt agaagaaaat
     5521 ccaggaccga tggccaagcc tttgtctcaa gaagaatcca ccctcattga aagagcaacg
     5581 gctacaatca acagcatccc catctctgaa gattacagcg tcgccagcgc agctctctct
     5641 agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg accttgtgca
     5701 gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc
     5761 gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg acaggtgctt
     5821 ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca gccgacggca
     5881 gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagg tcacccgcgc
     5941 gcggactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     6001 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     6061 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     6121 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg ttcgaaccag
     6181 tacgatggtt ttccccgtat ccccccaggt gtctgcaggc tcaaagagca gcgagaagcg
     6241 ttcagaggaa agcgatcccg tgccaccttc cccgtgcccg ggctgtcccc gcacgctgcc
     6301 ggctcgggga tgcgggggga gcgccggacc ggagcggagc cccgggcggc tcgctgctgc
     6361 cccctagcgg gggagggacg taattacatc cctgggggct ttgggggggg gctgtccctg
     6421 ataaaagttt tgttacttta tagaagaaat tttgagtttt tgtttttttt taataaataa
     6481 ataaacataa ataaattgtt tgttgaattt attattagta tgtaagtgta aatataataa
     6541 aacttaatat ctattcaaat taataaataa acctcgatat acagaccgat aaaacacatg
     6601 cgtcaatttt acgcatgatt atctttaacg tacgtcacaa tatgattatc tttctagggt
     6661 taatctagct gcggcgcgcc atcaggatca tatcgtcggg tcttttttcc ggctcagtca
     6721 tcgcccaagc tggcgctatc tgggcatcgg ggaggaagaa gcccgtgcct tttcccgcga
     6781 ggttgaagcg gcatggaaag agtttgccga ggatgactgc tgctgcattg acgttgagcg
     6841 aaaacgcacg tttaccatga tgattcggga aggtgtggcc atgcacgcct ttaacggtga
     6901 actgttcgtt caggccacct gggataccag ttcgtcgcgg cttttccgga cacagttccg
     6961 gatggtcagc ccgaagcgca tcagcaaccc gaacaatacc ggcgacagcc ggaactgccg
     7021 tgccggtgtg cagattaatg acagcggtgc ggcgctggga tattacgtca gcgaggacgg
     7081 gtatcctggc tggatgccgc agaaatggac atggataccc cgtgagttac ccggcgggcg
     7141 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     7201 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     7261 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     7321 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     7381 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     7441 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     7501 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     7561 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     7621 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     7681 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7741 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7801 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7861 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7921 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7981 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     8041 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     8101 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     8161 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     8221 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     8281 tcaatctaaa gtatatatga gtaaacttgg tctgacagtc agaagaactc gtcaagaagg
     8341 cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg
     8401 tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga
     8461 tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc
     8521 accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc
     8581 atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc
     8641 agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt
     8701 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca
     8761 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc
     8821 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct
     8881 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca
     8941 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc
     9001 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc
     9061 ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat catactcttc
     9121 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     9181 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     9241 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     9301 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     9361 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     9421 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     9481 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     9541 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     9601 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     9661 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     9721 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     9781 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     9841 gtaaaacgac ggccagtgag cgcgcctcgt tcattcacgt ttttgaaccc gtggaggacg
     9901 ggcagactcg cggtgcaaat gtgttttaca gcgtgatgga gcagatgaag atgctcgaca
     9961 cgctgcagaa cacgagctcc accgcggggc gcgcc
//
