LOCUS       sequence-432518-       10741 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable, inducible expression in cytoplasm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10741)
  AUTHORS   Zhong X, Li Q, Polacco BJ, Patil T, DiBerto JF, Vartak R, Xu J, 
            Marley A, Foussard H, Roth BL, Eckhardt M, Von Zastrow M, Krogan NJ,
            Huttenhain R
  TITLE     An automated proximity proteomics pipeline for subcellular proteome 
            and protein interaction mapping.
  JOURNAL   bioRxiv. 2023 Apr 12:2023.04.11.536358. doi: 
            10.1101/2023.04.11.536358. Preprint.
  PUBMED    37090610
REFERENCE   2  (bases 1 to 10741)
  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"; journalName: "bioRxiv. 
            2023 Apr 12:2023.04.11.536358. doi: 10.1101/2023.04.11.536358. 
            Preprint."
COMMENT     Sequence Label: Cyto-APEX2
FEATURES             Location/Qualifiers
     source          1..10741
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             110..136
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     polyA_signal    323..457
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    501..616
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     CDS             complement(879..923)
                     /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    1108..1341
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1838..1963
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             2010..2643
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    2811..2945
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        2993..3097
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3098..3166,3167..3958)
                     /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(3316..3335)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4129..4717
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4618..4637
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4871..4888
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5005..5026
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5041..5046,5047..5064,5065..5071)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5079..5095
                     /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     5084..5106
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5103..5119
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5103..5119
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     LTR             5247..5880
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    6087..6675
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6193..6204
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6602..6622
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6689..7288)
                     /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(6772..6792)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7269..7288
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(7297..7626)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7311..7446
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             complement(7637..8383)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     promoter        complement(8402..8912)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8465..8483)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     promoter        8929..9296
                     /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"
     protein_bind    8937..8955
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8973..8991
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9009..9027
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9045..9063
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9081..9099
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9117..9135
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9153..9171
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             join(9306..9308,9309..9311,9312..10022)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(9306..9308,9309..9311,9312..10022)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(9351..9372)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(9612..9631)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9959..9980
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      10065..10074
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             10074..10097
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(10098..10741,1..103)
                     /codon_start=1
                     /product="soybean ascorbate peroxidase, engineered to serve
                     as a monomeric tag for electron microscopy and live-cell 
                     proteomics (Lam et al., 2015)"
                     /label=APEX2
                     /note="more active than the original APEX due to the A134P 
                     mutation"
                     /translation="GKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAGT
                     FDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVAV
                     EVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGHT
                     IGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVDK
                     YAADEDAFFADYAEAHQKLSELGFADA"
ORIGIN
        1 attccgccct ctcgttgaca aatatgcagc ggacgaagat gccttctttg ctgattacgc
       61 tgaggctcac caaaagcttt ccgagcttgg gtttgctgat gccctgcagc tgcctcccct
      121 ggagcgcctg accctggact aaggatccag accacctccc ctgcgagcta agctggacag
      181 ccaatgacgg gtaagagagt gacatttttc actaacctaa gacaggaggg ccgtcagagc
      241 tactgcctaa tccaaagacg ggtaaaagtg ataaaaatgt atcactccaa cctaagacag
      301 gcgcagcttc cgagggattt gagatccaga catgataaga tacattgatg agtttggaca
      361 aaccaaaact agaatgcagt gaaaaaaatg ccttatttgt gaaatttgtg atgctattgc
      421 cttatttgta accattataa gctgcaataa acaagttaag tcgataaact ggatctctgc
      481 tgtccctgta ataaacccga aaattttgaa tttttgtaat ttgtttttgt aattctttag
      541 tttgtatgtc tgttgctatt atgtctacta ttctttcccc tgcactgtac cccccaatcc
      601 ccccttttct tttaaaggcg ataccgtcga gatccgttca ctaatcgaat ggatctgtct
      661 ctgtctctct ctccaccttc ttcttctatt ccttcgggcc tgtcgggtcc cctcggggtt
      721 gggaggtggg tctgaaacga taatggtgaa tatccctgcc taactctatt cactatagaa
      781 agtacagcaa aaactattct taaacctacc aagcctccta ctatcattat gaataatttt
      841 atataccaca gccaatttgt tatgttaaac caattccaca aacttgccca tttatctaat
      901 tccaataatt cttgttcatt cttttcttgc tggttttgcg attcttcaat taaggagtgt
      961 attaagcttg tgtaattgtt aatttctctg tcccactcca tccaggtcgt gtgattccaa
     1021 atctgttcca gagatttatt actccaacta gcattccaag gcacagcagt ggtgcaaatg
     1081 agttttccag agcaacccca aatccccagg agctgttgat cctttaggta tctttccaca
     1141 gccaggattc ttgcctggag ctgcttgatg ccccagactg tgagttgcaa cagatgctgt
     1201 tgcgcctcaa tagccctcag caaattgttc tgctgctgca ctataccaga caataattgt
     1261 ctggcctgta ccgtcagcgt cattgacgct gcgcccatag tgcttcctgc tgctcccaag
     1321 aacccaagga acaaagctcc tattcccact gctctttttt ctctctgcac cactcttctc
     1381 tttgccttgg tgggtgctac tcctaatggt tcaattttta ctactttata tttatataat
     1441 tcacttctcc aattgtccct catatctcct cctccaggtc tgaagatcag cggccggccg
     1501 cttgctgtgc ggtggtctta cttttgtttt gctcttcctc tatcttgtct aaagcttcct
     1561 tggtgtcttt tatctctatc ctttgatgca cacaatagag ggttgctact gtattatata
     1621 atgatctaag ttcttctgat cctgtctgaa gggatggttg tagctgtccc agtatttgtc
     1681 tacagccttc tgatgtttct aacaggccag gattaactgc gaatcgttct agctccctgc
     1741 ttgcccatac tatatgtttt aatttatatt ttttctttcc ccctggcctt aaccgaattt
     1801 tttcccatcg cgatctaatt ctcccccgct taatactgac gctctcgcac ccatctctct
     1861 ccttctagcc tccgctagtc aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc
     1921 ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga gagctcctct ggtttccctt
     1981 tcgctttcaa gtccctgttc gggcgccact gctagagatt ttccacactg actaaaaggg
     2041 tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta cttgaagcac
     2101 tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag agagctccca
     2161 ggctcagatc tggtctaacc agagagaccc agtacaggca aaaagcagct gcttatatgc
     2221 aggatctgag ggctcgccac tccccagtcc cgcccaggcc acgcctccct ggaaagtccc
     2281 cagcggaaag tcccttgtag caagctcgat atcagcagtt cttgaagtac tccggatgca
     2341 gctctcgggc cacgtgatga aatgctaggc ggctgtcaaa cctccactct aacacttctc
     2401 tctccgggtc atccatccca tgcaggctca cagggtgtaa caagctggtg ttctctcctt
     2461 tattggcctc ttctacctta tctggctcaa ctggtactag cttgtagcac catccaaagg
     2521 tcagtggata tctgacccct ggccctggtg tgtagttctg ctaatcaggg aagtagcctt
     2581 gtgtgtggta gatccacaga tcaaggatat cttgtcttct ttgggagtga attagccctt
     2641 ccaactacta agtttgtagt acatatttaa caaatacaat ttctttaaaa tgaaaataat
     2701 tcagaggaat cacaggttta gagtaaatga aaccacaggt aattggcagt ggtaataggg
     2761 tatggggtgg gaagtttggg atgattttgg ttagcttgag ttatccagtt gatccagaca
     2821 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     2881 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     2941 aagttgatct cccgatccgt cgacgtcagg tggcactttt cggggaaatg tgcgcggaac
     3001 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3061 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3121 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3181 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3241 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3301 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3361 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3421 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3481 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3541 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     3601 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     3661 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     3721 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     3781 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     3841 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     3901 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     3961 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4021 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4081 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4141 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4201 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4261 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4321 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4381 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4441 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4501 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4561 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4621 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4681 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4741 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4801 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     4861 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     4921 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     4981 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     5041 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     5101 cacaggaaac agctatgacc atgattacgc caagctctag ctagaggtcg acggtaaacg
     5161 ctagcaatgt caaggcctct cactctctga tattcatttc tttgcaagtt ataaatactg
     5221 aataataaga tgacatgaac tactactgct agagattttc cacactgact aaaagggtct
     5281 gagggatctc tagttaccag agtcacacaa cagacgggca cacactactt gaagcactca
     5341 aggcaagctt tattgaggct taagcagtgg gttccctagt tagccagaga gctcccaggc
     5401 tcagatctgg tctaaccaga gagacccagt acaggcaaaa agcagctgct tatatgcagg
     5461 atctgagggc tcgccactcc ccagtcccgc ccaggccacg cctccctgga aagtccccag
     5521 cggaaagtcc cttgtagcaa gctcgatatc agcagttctt gaagtactcc ggatgcagct
     5581 ctcgggccac gtgatgaaat gctaggcggc tgtcaaacct ccactctaac acttctctct
     5641 ccgggtcatc catcccatgc aggctcacag ggtgtaacaa gctggtgttc tctcctttat
     5701 tggcctcttc taccttatct ggctcaactg gtactagctt gtagcaccat ccaaaggtca
     5761 gtggatatct gacccctggc cctggtgtgt agttctgcta atcagggaag tagccttgtg
     5821 tgtggtagat ccacagatca aggatatctt gtcttcgttg ggagtgaatt agcccttcca
     5881 gtcccctctt ttcttttaaa aagtggctaa gatctacagc tgccttgtaa gtcattggtc
     5941 ttaaaggtac ctgaggtgtg actggaaaac ccacctcctc ctcctcttgt gcttctagcc
     6001 aggcacaatc agcattggta gctgctgtat tgctacttgt gattgctcca tgtttttcta
     6061 ggtctcgact gcagaattaa ttccaggcgg ggaggcggcc caaagggaga tccgactcgt
     6121 ctgagggcga aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg
     6181 aaggtccgct ggattgaggg ccgaagggac gtagcagaag gacgtcccgc gcagaatcca
     6241 ggtggcaaca caggcgagca gccatggaaa ggacgtcagc ttccccgaca acaccacgga
     6301 attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag
     6361 ttccgccgtg gcaataggga gggggaaagc gaaagtcccg gaaaggagct gacaggtggt
     6421 ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca gtgcacacca cgccacgttg
     6481 cctgacaacg ggccacaact cctcataaag agacagcaac caggatttat acaaggagga
     6541 gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc
     6601 acatagcgta aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta
     6661 atccagaggt tgattgttcc agacgcgttc aggcaccggg cttgcgggtc atgcaccagg
     6721 tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt gaagccgagc cgctcgtaga
     6781 aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg caccccggcg cgctcggccg
     6841 cctccactcc ggggagcacg acggcgctgc ccagaccctt gccctggtgg tcgggcgaga
     6901 cgccgacggt ggccaggaac cacgcgggct ccttgggccg gtgcggcgcc aggaggcctt
     6961 ccatctgttg ctgcgcggcc agccgggaac cgctcaactc ggccatgcgc gggccgatct
     7021 cggcgaacac cgcccccgct tcgacgctct ccggcgtggt ccagaccgcc accgcggcgc
     7081 cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac gcgcgtgagg aagagttctt
     7141 gcagctcggt gacccgctcg atgtggcggt ccgggtcgac ggtgtggcgc gtggcggggt
     7201 agtcggcgaa cgcggcggcg agggtgcgta cggcccgggg gacgtcgtcg cgggtggcga
     7261 ggcgcaccgt gggcttgtac tcggtcatgg tcgcgttttg caaaagccta ggcctccaaa
     7321 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     7381 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     7441 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     7501 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     7561 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     7621 acacacattc gcgcgtttac ccggggagca tgtcaaggtc aaaatcgtca agagcgtcag
     7681 caggcagcat atcaaggtca aagtcgtcaa gggcatcggc tgggagcatg tctaagtcaa
     7741 aatcgtcaag ggcgtcggtc ggcccgccgc tttcgcactt tagctgtttc tccaggccac
     7801 atatgattag ttccaggccg aaaaggaagg caggttcggc tccctgccgg tcgaacagct
     7861 caattgcttg tttcagaagt gggggcatag aatcggtggt aggtgtctct ctttcctctt
     7921 ttgctacttg atgctcctgt tcctccaata cgcagcccag tgtaaagtgg cccacggcgg
     7981 acagagcgta cagtgcgttc tccagggaga agccttgctg acacaggaac gcgagctgat
     8041 tttccagggt ttcgtactgt ttctctgttg ggcgggtgcc gagatgcact ttagccccgt
     8101 cgcgatgtga gaggagagca cagcggtatg acttggcgtt gttccgcaga aagtcttgcc
     8161 atgactcgcc ttccaggggg caggagtggg tatgatgcct gtccagcatc tcgattggca
     8221 gggcatcgag cagggcccgc ttgttcttca cgtgccagta cagggtaggc tgctcaactc
     8281 ccagcttttg agcgagtttc cttgtcgtca ggccttcgat accgactcca ttgagtaatt
     8341 ccagagcaga gtttatgact ttgctcttgt ccagtctaga catggtaatt cgatgatccc
     8401 cctggggaga gaggtcggtg attcggtcaa cgagggagcc gactgccgac gtgcgctccg
     8461 gaggcttgca gaatgcggaa caccgcgcgg gcaggaacag ggcccacact accgccccac
     8521 accccgcctc ccgcaccgcc ccttcccggc cgctgctctc ggcgcgccct gctgagcagc
     8581 cgctattggc cacagcccat cgcggtcggc gcgctgccat tgctccctgg cgctgtccgt
     8641 ctgcgagggt actagtgaga cgtgcggctt ccgtttgtca cgtccggcac gccgcgaacc
     8701 gcaaggaacc ttcccgactt aggggcggag caggaagcgt cgccgggggg cccacaaggg
     8761 tagcggcgaa gatccgggtg acgctgcgaa cggacgtgaa gaatgtgcga gacccagggt
     8821 cggcgccgct gcgtttcccg gaaccacgcc cagagcagcc gcgtccctgc gcaaacccag
     8881 ggctgccttg gaaaaggcgc aaccccaacc ccgtggaatt atcacctcga gtttactccc
     8941 tatcagtgat agagaacgta tgaagagttt actccctatc agtgatagag aacgtatgca
     9001 gactttactc cctatcagtg atagagaacg tataaggagt ttactcccta tcagtgatag
     9061 agaacgtatg accagtttac tccctatcag tgatagagaa cgtatctaca gtttactccc
     9121 tatcagtgat agagaacgta tatccagttt actccctatc agtgatagag aacgtataag
     9181 ctttgcttat gtaaaccagg gcgcctataa aagagtgctg attttttgag taaacttcaa
     9241 ttccacaaca cttttgtctt ataccaactt tccgtaccac ttcctaccct cgtaaagaat
     9301 tcaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     9361 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     9421 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     9481 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     9541 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     9601 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     9661 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     9721 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     9781 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     9841 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     9901 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     9961 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
    10021 agtccggact cagaactcga gctcaagctt cgaattctgc ggccgccacc atggactaca
    10081 aggatgacga cgataaggga aagtcttacc caactgtgag tgctgattac caggacgccg
    10141 ttgagaaggc gaagaagaag ctcagaggct tcatcgctga gaagagatgc gctcctctaa
    10201 tgctccgttt ggcattccac tctgctggaa cctttgacaa gggcacgaag accggtggac
    10261 ccttcggaac catcaagcac cctgccgaac tggctcacag cgctaacaac ggtcttgaca
    10321 tcgctgttag gcttttggag ccactcaagg cggagttccc tattttgagc tacgccgatt
    10381 tctaccagtt ggctggcgtt gttgccgttg aggtcacggg tggacctaag gttccattcc
    10441 accctggaag agaggacaag cctgagccac caccagaggg tcgcttgccc gatcccacta
    10501 agggttctga ccatttgaga gatgtgtttg gcaaagctat ggggcttact gaccaagata
    10561 tcgttgctct atctgggggt cacactattg gagctgcaca caaggagcgt tctggatttg
    10621 agggtccctg gacctctaat cctcttattt tcgacaactc atacttcacg gagttgttga
    10681 gtggtgagaa ggaaggtctc cttcagctac cttctgacaa ggctcttttg tctgaccctg
    10741 t
//
