LOCUS       sequence-268081-       11922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2nd generation lentiviral vector for over expressing human PD-L1 WT 
            with PD-L1 shRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11922)
  AUTHORS   Li CW, Lim SO, Xia W, Lee HH, Chan LC, Kuo CW, Khoo KH, Chang SS, 
            Cha JH, Kim T, Hsu JL, Wu Y, Hsu JM, Yamaguchi H, Ding Q, Wang Y, 
            Yao J, Lee CC, Wu HJ, Sahin AA, Allison JP, Yu D, Hortobagyi GN, 
            Hung MC
  TITLE     Glycosylation and stabilization of programmed death ligand-1 
            suppresses T-cell activity.
  JOURNAL   Nat Commun. 2016 Aug 30;7:12632. doi: 10.1038/ncomms12632.
  PUBMED    27572267
REFERENCE   2  (bases 1 to 11922)
  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: "Nat 
            Commun."; date: "2016-08-30"; volume: "7:12632. doi"; pages: " 
            10.1038/ncomms12632"
COMMENT     Sequence Label: pGIPz-PD-L1 WT
FEATURES             Location/Qualifiers
     source          1..11922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2063..2180
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(2245..2619)
                     /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      2616..2625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2629..2647)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2639..2686)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     enhancer        2791..3094
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3095..3298
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3249..3269
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3295..3319
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4266..4289
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    4328..4900
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4495..4512)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4722..4741
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4903..5505
                     /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     5402..5422
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     ncRNA           5512..5633
                     /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           5661..5675
                     /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           5703..5829
                     /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    5897..6485
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5950..5970)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6368..6379)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6692..6925
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7026..7043)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7032..7256
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7302..7730
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7389..7408)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7599..7620
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7739..7759)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7744..8073
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7924..8059
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7986..8005
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             8122..9147
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             8122..9147
                     /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    9277..9398
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9447..9463)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9447..9463)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9460..9482)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9471..9487
                     /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(9495..9501,9502..9519,9520..9525))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9540..9561
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9678..9695)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9849..10437)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9929..9948)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10608..11399,11400..11468))
                     /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     11231..11250
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11469..11573)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    11621..11755
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
     2041 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     2101 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     2161 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag
     2221 taccgggccc gctctagtcc ggaatcagtc ctgctcctcg gccacgaagt gcacgcagtt
     2281 gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat
     2341 ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag
     2401 ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg
     2461 gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac
     2521 gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg
     2581 cgcggtgagc accggaacgg cactggtcaa cttggccatg gtggccctcc tatagtgagt
     2641 cgtattatac tatgccgata tactatgccg atgattaatt gtcaacacgt gctgcaggtc
     2701 cgaggttcta gacgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
     2761 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     2821 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     2881 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     2941 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     3001 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     3061 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     3121 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     3181 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     3241 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
     3301 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag
     3361 acaccgactc tactagagga tctgccacca tcgatatgag gatatttgct gtctttatat
     3421 tcatgaccta ctggcatttg ctgaacgcat ttactgtcac ggttcccaag gacctatatg
     3481 tggtagagta tggtagcaat atgacaattg aatgcaaatt cccagtagaa aaacaattag
     3541 acctggctgc actaattgtc tattgggaaa tggaggataa gaacattatt caatttgtgc
     3601 atggagagga agacctgaag gttcagcata gtagctacag acagagggcc cggctgttga
     3661 aggaccagct ctccctggga aatgctgcac ttcagatcac agatgtgaaa ttgcaggatg
     3721 caggggtgta ccgctgcatg atcagctatg gtggtgccga ctacaagcga attactgtga
     3781 aagtcaatgc cccatacaac aaaatcaacc aaagaatttt ggttgtggat ccagtcacct
     3841 ctgaacatga actgacatgt caggctgagg gctaccccaa ggccgaagtc atctggacaa
     3901 gcagtgacca tcaagtcctg agtggtaaga ccaccaccac caattccaag agagaggaga
     3961 agcttttcaa tgtgaccagc acactgagaa tcaacacaac aactaatgag attttctact
     4021 gcacttttag gagattagat cctgaggaaa accatacagc tgaattggtc atcccagaac
     4081 tacctctggc acatcctcca aatgaaagga ctcacttggt aattctggga gccatcttat
     4141 tatgccttgg tgtagcactg acattcatct tccgtttaag aaaagggaga atgatggatg
     4201 tgaaaaaatg tggcatccaa gatacaaact caaagaagca aagtgataca catttggagg
     4261 agacggacta caaagacgat gacgacaagt gagctagcta atgtacaagt agcggccgca
     4321 aattccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     4381 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     4441 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     4501 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     4561 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     4621 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     4681 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     4741 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     4801 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     4861 accacgggga cgtggttttc ctttgaaaaa cacgataata ccatggccac cgagtacaag
     4921 cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc
     4981 gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg
     5041 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     5101 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     5161 gcggtgttcg ccgagatcgg ctcgcgcatg gccgagttga gcggttcccg gctggccgcg
     5221 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     5281 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     5341 cccggagtgg aggcggccga gcgcgctggg gtgcccgcct tcctggagac ctccgcgccc
     5401 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     5461 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtttg tttgaatgag
     5521 gcttcagtac tttacagaat cgttgcctgc acatcttgga aacacttgct gggattactt
     5581 cttcaggtta acccaacaga aggctcgaga aggtatattg ctgttgacag tgagcgcatg
     5641 aagaaagatg gagtcaaata gtgaagccac agatgtattt gactccatct ttcttcattt
     5701 gcctactgcc tcggaattca aggggctact ttaggagcaa ttatcttgtt tactaaaact
     5761 gaataccttg ctatctcttt gatacatttt tacaaagctg aattaaaatg gtataaatta
     5821 aatcactttt ttggccggcc aacttgcagt acaccggacc acgaccctag gatcgacgcg
     5881 gacacgcgtc tggaacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     5941 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     6001 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     6061 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     6121 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     6181 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     6241 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     6301 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     6361 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     6421 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     6481 cccgcctgga attaattctg cagtcgagac ctagaaaaac atggagcaat cacaagtagc
     6541 aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg
     6601 ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat
     6661 cttagccact ttttaaaaga aaagagggga ctggaagggc taattcactc ccaacgaaga
     6721 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     6781 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     6841 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     6901 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     6961 taacttgcaa agaaatgaat atcagagagt gagaggcctt gacattgttt aaacccgctg
     7021 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     7081 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     7141 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     7201 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     7261 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     7321 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     7381 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     7441 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     7501 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     7561 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     7621 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     7681 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     7741 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     7801 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     7861 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     7921 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     7981 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     8041 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     8101 ttcggatctg atcagcacgt gatgaaaaag cctgaactca ccgcgacgtc tgtcgagaag
     8161 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     8221 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     8281 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
     8341 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc
     8401 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
     8461 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
     8521 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
     8581 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
     8641 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
     8701 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
     8761 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc
     8821 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
     8881 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa
     8941 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
     9001 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
     9061 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc
     9121 agcactcgtc cgagggcaaa ggaatagcac gtgctacgag atttcgattc caccgccgcc
     9181 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     9241 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     9301 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     9361 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     9421 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     9481 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     9541 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     9601 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     9661 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     9721 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     9781 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     9841 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     9901 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     9961 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
    10021 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
    10081 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
    10141 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
    10201 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
    10261 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
    10321 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
    10381 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
    10441 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
    10501 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
    10561 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
    10621 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
    10681 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
    10741 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
    10801 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
    10861 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
    10921 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
    10981 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
    11041 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
    11101 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
    11161 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
    11221 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
    11281 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
    11341 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
    11401 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
    11461 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
    11521 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
    11581 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc aacttgttta ttgcagctta
    11641 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
    11701 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg
    11761 gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc
    11821 caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta
    11881 aagaaattgt atttgttaaa tatgtactac aaacttagta gt
//
