LOCUS       sequence-236077-       11922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human PD-L1 4NQ 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 4NQ
FEATURES             Location/Qualifiers
     source          1..11922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        84..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        388..591
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     542..562
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     588..612
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1559..1582
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1621..2193
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1788..1805)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2015..2034
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2196..2798
                     /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     2695..2715
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     ncRNA           2805..2926
                     /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           2954..2968
                     /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           2996..3122
                     /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    3190..3778
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3243..3263)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3661..3672)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3985..4218
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(4319..4336)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4325..4549
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4595..5023
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4682..4701)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4892..4913
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5032..5052)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5037..5366
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5217..5352
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5279..5298
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5415..6440
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             5415..6440
                     /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    6570..6691
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6740..6756)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6740..6756)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6753..6775)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6764..6780
                     /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(6788..6794,6795..6812,6813..6818))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6833..6854
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6971..6988)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7142..7730)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7222..7241)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7901..8692,8693..8761))
                     /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     8524..8543
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8762..8866)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    8914..9048
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             9216..9849
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    9896..10021
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10518..10751
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10936..10980
                     /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    11278..11395
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(11460..11834)
                     /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      11831..11840
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(11844..11862)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(11854..11901)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
ORIGIN
        1 ctagacgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      421 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      481 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      541 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
      601 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccga
      661 ctctactaga ggatctgcca ccatcgatat gaggatattt gctgtcttta tattcatgac
      721 ctactggcat ttgctgaacg catttactgt cacggttccc aaggacctat atgtggtaga
      781 gtatggtagc caaatgacaa ttgaatgcaa attcccagta gaaaaacaat tagacctggc
      841 tgcactaatt gtctattggg aaatggagga taagaacatt attcaatttg tgcatggaga
      901 ggaagacctg aaggttcagc atagtagcta cagacagagg gcccggctgt tgaaggacca
      961 gctctccctg ggaaatgctg cacttcagat cacagatgtg aaattgcagg atgcaggggt
     1021 gtaccgctgc atgatcagct atggtggtgc cgactacaag cgaattactg tgaaagtcaa
     1081 tgccccatac aacaaaatca accaaagaat tttggttgtg gatccagtca cctctgaaca
     1141 tgaactgaca tgtcaggctg agggctaccc caaggccgaa gtcatctgga caagcagtga
     1201 ccatcaagtc ctgagtggta agaccaccac caccaattcc aagagagagg agaagctttt
     1261 ccaagtgacc agcacactga gaatccagac aacaactaat gagattttct actgcacttt
     1321 taggagatta gatcctgagg aacagcatac agctgaattg gtcatcccag aactacctct
     1381 ggcacatcct ccaaatgaaa ggactcactt ggtaattctg ggagccatct tattatgcct
     1441 tggtgtagca ctgacattca tcttccgttt aagaaaaggg agaatgatgg atgtgaaaaa
     1501 atgtggcatc caagatacaa actcaaagaa gcaaagtgat acacatttgg aggagacgga
     1561 ctacaaagac gatgacgaca agtgagctag ctaatgtaca agtagcggcc gcaaattccg
     1621 cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
     1681 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
     1741 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
     1801 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
     1861 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
     1921 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
     1981 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
     2041 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
     2101 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
     2161 ggacgtggtt ttcctttgaa aaacacgata ataccatggc caccgagtac aagcccacgg
     2221 tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg
     2281 ccgactaccc cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg
     2341 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     2401 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     2461 tcgccgagat cggctcgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     2521 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     2581 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     2641 tggaggcggc cgagcgcgct ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     2701 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     2761 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt ttgtttgaat gaggcttcag
     2821 tactttacag aatcgttgcc tgcacatctt ggaaacactt gctgggatta cttcttcagg
     2881 ttaacccaac agaaggctcg agaaggtata ttgctgttga cagtgagcgc atgaagaaag
     2941 atggagtcaa atagtgaagc cacagatgta tttgactcca tctttcttca tttgcctact
     3001 gcctcggaat tcaaggggct actttaggag caattatctt gtttactaaa actgaatacc
     3061 ttgctatctc tttgatacat ttttacaaag ctgaattaaa atggtataaa ttaaatcact
     3121 tttttggccg gccaacttgc agtacaccgg accacgaccc taggatcgac gcggacacgc
     3181 gtctggaaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     3241 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     3301 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     3361 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     3421 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     3481 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3541 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     3601 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3661 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3721 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     3781 ggaattaatt ctgcagtcga gacctagaaa aacatggagc aatcacaagt agcaatacag
     3841 cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc
     3901 cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     3961 actttttaaa agaaaagagg ggactggaag ggctaattca ctcccaacga agacaagatc
     4021 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     4081 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     4141 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     4201 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     4261 caaagaaatg aatatcagag agtgagaggc cttgacattg tttaaacccg ctgatcagcc
     4321 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     4381 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     4441 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     4501 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     4561 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     4621 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4681 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4741 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4801 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4861 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4921 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4981 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     5041 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5101 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5161 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5221 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5281 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     5341 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     5401 ctgatcagca cgtgatgaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga
     5461 tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg
     5521 ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg
     5581 gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg
     5641 aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac
     5701 agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg
     5761 cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat
     5821 tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg
     5881 atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc
     5941 aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc
     6001 acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg
     6061 actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga
     6121 ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc
     6181 ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc
     6241 agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa
     6301 tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg
     6361 tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc
     6421 gtccgagggc aaaggaatag cacgtgctac gagatttcga ttccaccgcc gccttctatg
     6481 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     6541 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     6601 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6661 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     6721 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6781 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6841 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6901 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6961 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     7021 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     7081 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7141 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7201 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7261 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7321 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7381 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7441 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7501 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7561 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     7621 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7681 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7741 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7801 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7861 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7921 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7981 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     8041 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     8101 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     8161 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     8221 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8281 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8341 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8401 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8461 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8521 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8581 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8641 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8701 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8761 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8821 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8881 aagtgccacc tgacgtcgac ggatcgggag atcaacttgt ttattgcagc ttataatggt
     8941 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     9001 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact
     9061 caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac
     9121 ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat
     9181 tgtatttgtt aaatatgtac tacaaactta gtagttggaa gggctaattc actcccaaag
     9241 aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca
     9301 gaactacaca ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct
     9361 agtaccagtt gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt
     9421 acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt
     9481 tgacagccgc ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa
     9541 ctgctgatat cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg
     9601 cctgggcggg actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg
     9661 cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     9721 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     9781 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     9841 tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc
     9901 tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt
     9961 gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt
    10021 cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg
    10081 aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc
    10141 agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca
    10201 accatccctt cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct
    10261 ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag acaagataga
    10321 ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccggccgct gatcttcaga
    10381 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
    10441 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
    10501 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
    10561 atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
    10621 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
    10681 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
    10741 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
    10801 tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg
    10861 gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg
    10921 caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg
    10981 tggaattggt ttaacataac aaattggctg tggtatataa aattattcat aatgatagta
    11041 ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg
    11101 cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg
    11161 cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg
    11221 aacggatcgg cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt
    11281 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
    11341 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt
    11401 ttattacagg gacagcagag atccagtttg gttagtaccg ggcccgctct agtccggaat
    11461 cagtcctgct cctcggccac gaagtgcacg cagttgccgg ccgggtcgcg cagggcgaac
    11521 tcccgccccc acggctgctc gccgatctcg gtcatggccg gcccggaggc gtcccggaag
    11581 ttcgtggaca cgacctccga ccactcggcg tacagctcgt ccaggccgcg cacccacacc
    11641 caggccaggg tgttgtccgg caccacctgg tcctggaccg cgctgatgaa cagggtcacg
    11701 tcgtcccgga ccacaccggc gaagtcgtcc tccacgaagt cccgggagaa cccgagccgg
    11761 tcggtccaga actcgaccgc tccggcgacg tcgcgcgcgg tgagcaccgg aacggcactg
    11821 gtcaacttgg ccatggtggc cctcctatag tgagtcgtat tatactatgc cgatatacta
    11881 tgccgatgat taattgtcaa cacgtgctgc aggtccgagg tt
//
