LOCUS       sequence-234833-       12666 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express PD-L1-EGFP fusion protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12666)
  AUTHORS   Yang Y, Li CW, Chan LC, Wei Y, Hsu JM, Xia W, Cha JH, Hou J, Hsu JL,
            Sun L, Hung MC
  TITLE     Exosomal PD-L1 harbors active defense function to suppress T cell 
            killing of breast cancer cells and promote tumor growth.
  JOURNAL   Cell Res. 2018 Aug;28(8):862-864. doi: 10.1038/s41422-018-0060-4. 
            Epub 2018 Jun 29.
  PUBMED    29959401
REFERENCE   2  (bases 1 to 12666)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41422-018-0060-4"; journalName: "Cell Res"; date: 
            "2018-08"; volume: "28"; issue: "8"; pages: "862-864"
COMMENT     Sequence Label: pGIPZ-PD-L1-EGFP
FEATURES             Location/Qualifiers
     source          1..12666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             155..788
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    835..960
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1457..1690
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1875..1919
                     /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    2217..2334
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(2399..2773)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     regulatory      2770..2779
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2783..2801)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2793..2840)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     enhancer        2945..3248
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3249..3452
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3403..3423
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3449..3473
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4465..4474
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4471..4473,4474..4476,4477..5190)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4471..4473,4474..4476,4477..5187)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4516..4537)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4777..4796)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5124..5145
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5226..5798
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5393..5410)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5620..5639
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5801..6403
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     6300..6320
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     ncRNA           6410..6531
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           6559..6573
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           6601..6727
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     misc_feature    6795..7383
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6848..6868)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7266..7277)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7590..7823
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7924..7941)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7930..8154
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8200..8628
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8287..8306)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8497..8518
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8637..8657)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8642..8971
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8822..8957
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8884..8903
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             9020..10045
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             9020..10045
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    10175..10296
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10345..10361)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10345..10361)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10358..10380)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10369..10385
                     /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(join(10393..10399,10400..10417,10418..10423))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10438..10459
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10576..10593)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10747..11335)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10827..10846)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11506..12297,12298..12366))
                     /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     12129..12148
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12367..12471)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    12519..12653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 aagctaacca aaatcatccc aaacttccca ccccataccc tattaccact gccaattacc
       61 tgtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt
      121 gtatttgtta aatatgtact acaaacttag tagttggaag ggctaattca ctcccaaaga
      181 agacaagata tccttgatct gtggatctac cacacacaag gctacttccc tgattagcag
      241 aactacacac cagggccagg ggtcagatat ccactgacct ttggatggtg ctacaagcta
      301 gtaccagttg agccagataa ggtagaagag gccaataaag gagagaacac cagcttgtta
      361 caccctgtga gcctgcatgg gatggatgac ccggagagag aagtgttaga gtggaggttt
      421 gacagccgcc tagcatttca tcacgtggcc cgagagctgc atccggagta cttcaagaac
      481 tgctgatatc gagcttgcta caagggactt tccgctgggg actttccagg gaggcgtggc
      541 ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc tgctttttgc
      601 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
      661 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
      721 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
      781 ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct
      841 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg
      901 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc
      961 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga
     1021 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
     1081 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
     1141 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
     1201 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
     1261 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac
     1321 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
     1381 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
     1441 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
     1501 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
     1561 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
     1621 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
     1681 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
     1741 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
     1801 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     1861 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     1921 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     1981 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     2041 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     2101 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     2161 acggatcggc actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt
     2221 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     2281 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
     2341 tattacaggg acagcagaga tccagtttgg ttagtaccgg gcccgctcta gtccggaatc
     2401 agtcctgctc ctcggccacg aagtgcacgc agttgccggc cgggtcgcgc agggcgaact
     2461 cccgccccca cggctgctcg ccgatctcgg tcatggccgg cccggaggcg tcccggaagt
     2521 tcgtggacac gacctccgac cactcggcgt acagctcgtc caggccgcgc acccacaccc
     2581 aggccagggt gttgtccggc accacctggt cctggaccgc gctgatgaac agggtcacgt
     2641 cgtcccggac cacaccggcg aagtcgtcct ccacgaagtc ccgggagaac ccgagccggt
     2701 cggtccagaa ctcgaccgct ccggcgacgt cgcgcgcggt gagcaccgga acggcactgg
     2761 tcaacttggc catggtggcc ctcctatagt gagtcgtatt atactatgcc gatatactat
     2821 gccgatgatt aattgtcaac acgtgctgca ggtccgaggt tctagacgta ttaccgccat
     2881 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     2941 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     3001 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     3061 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     3121 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     3181 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     3241 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     3301 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     3361 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     3421 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga
     3481 gacgccatcc acgctgtttt gacctccata gaagacaccg actctactag aggatctgcc
     3541 accatcgata tgaggatatt tgctgtcttt atattcatga cctactggca tttgctgaac
     3601 gcatttactg tcacggttcc caaggaccta tatgtggtag agtatggtag caatatgaca
     3661 attgaatgca aattcccagt agaaaaacaa ttagacctgg ctgcactaat tgtctattgg
     3721 gaaatggagg ataagaacat tattcaattt gtgcatggag aggaagacct gaaggttcag
     3781 catagtagct acagacagag ggcccggctg ttgaaggacc agctctccct gggaaatgct
     3841 gcacttcaga tcacagatgt gaaattgcag gatgcagggg tgtaccgctg catgatcagc
     3901 tatggtggtg ccgactacaa gcgaattact gtgaaagtca atgccccata caacaaaatc
     3961 aaccaaagaa ttttggttgt ggatccagtc acctctgaac atgaactgac atgtcaggct
     4021 gagggctacc ccaaggccga agtcatctgg acaagcagtg accatcaagt cctgagtggt
     4081 aagaccacca ccaccaattc caagagagag gagaagcttt tcaatgtgac cagcacactg
     4141 agaatcaaca caacaactaa tgagattttc tactgcactt ttaggagatt agatcctgag
     4201 gaaaaccata cagctgaatt ggtcatccca gaactacctc tggcacatcc tccaaatgaa
     4261 aggactcact tggtaattct gggagccatc ttattatgcc ttggtgtagc actgacattc
     4321 atcttccgtt taagaaaagg gagaatgatg gatgtgaaaa aatgtggcat ccaagataca
     4381 aactcaaaga agcaaagtga tacacatttg gaggagacgg gaattctgca gtcgacggta
     4441 ccgcgggccc gggatccacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
     4501 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     4561 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     4621 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     4681 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     4741 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     4801 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     4861 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     4921 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     4981 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     5041 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     5101 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     5161 actctcggca tggacgagct gtacaagtaa gctagctaat gtacaagtag cggccgcaaa
     5221 ttccgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     5281 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     5341 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     5401 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     5461 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     5521 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     5581 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     5641 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     5701 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     5761 cacggggacg tggttttcct ttgaaaaaca cgataatacc atggccaccg agtacaagcc
     5821 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc
     5881 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt
     5941 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     6001 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     6061 ggtgttcgcc gagatcggct cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     6121 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     6181 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     6241 cggagtggag gcggccgagc gcgctggggt gcccgccttc ctggagacct ccgcgccccg
     6301 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     6361 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtttgtt tgaatgaggc
     6421 ttcagtactt tacagaatcg ttgcctgcac atcttggaaa cacttgctgg gattacttct
     6481 tcaggttaac ccaacagaag gctcgagaag gtatattgct gttgacagtg agcgcatgaa
     6541 gaaagatgga gtcaaatagt gaagccacag atgtatttga ctccatcttt cttcatttgc
     6601 ctactgcctc ggaattcaag gggctacttt aggagcaatt atcttgttta ctaaaactga
     6661 ataccttgct atctctttga tacattttta caaagctgaa ttaaaatggt ataaattaaa
     6721 tcactttttt ggccggccaa cttgcagtac accggaccac gaccctagga tcgacgcgga
     6781 cacgcgtctg gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     6841 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     6901 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     6961 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     7021 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     7081 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     7141 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     7201 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     7261 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     7321 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     7381 cgcctggaat taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa
     7441 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg
     7501 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
     7561 tagccacttt ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca
     7621 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     7681 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     7741 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     7801 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     7861 acttgcaaag aaatgaatat cagagagtga gaggccttga cattgtttaa acccgctgat
     7921 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     7981 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     8041 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     8101 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     8161 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     8221 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     8281 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     8341 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     8401 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     8461 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     8521 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     8581 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     8641 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     8701 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     8761 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     8821 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     8881 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     8941 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     9001 cggatctgat cagcacgtga tgaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt
     9061 tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc
     9121 tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc
     9181 cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat
     9241 tccggaagtg cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg
     9301 tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc
     9361 ggtcgcggag gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg
     9421 cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat
     9481 tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt
     9541 cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct
     9601 cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt
     9661 cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt
     9721 ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc
     9781 ggagcttgca ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact
     9841 ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga
     9901 cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc
     9961 ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag
    10021 cactcgtccg agggcaaagg aatagcacgt gctacgagat ttcgattcca ccgccgcctt
    10081 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
    10141 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
    10201 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
    10261 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
    10321 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
    10381 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
    10441 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
    10501 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
    10561 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
    10621 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
    10681 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
    10741 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
    10801 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
    10861 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
    10921 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
    10981 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
    11041 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
    11101 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
    11161 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
    11221 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
    11281 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
    11341 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
    11401 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
    11461 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
    11521 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
    11581 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
    11641 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
    11701 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
    11761 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
    11821 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
    11881 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
    11941 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
    12001 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
    12061 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
    12121 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
    12181 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
    12241 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
    12301 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
    12361 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
    12421 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
    12481 ccgaaaagtg ccacctgacg tcgacggatc gggagatcaa cttgtttatt gcagcttata
    12541 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
    12601 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga
    12661 taactc
//
