LOCUS       sequence-318425-       14709 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-inducible expression of Atg4B(C74A) fused with 
            mTurquoise2 to inhibit autophagy.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14709)
  AUTHORS   Yamamoto K, Venida A, Yano J, Biancur DE, Kakiuchi M, Gupta S, Sohn 
            ASW, Mukhopadhyay S, Lin EY, Parker SJ, Banh RS, Paulo JA, Wen KW, 
            Debnath J, Kim GE, Mancias JD, Fearon DT, Perera RM, Kimmelman AC
  TITLE     Autophagy promotes immune evasion of pancreatic cancer by degrading 
            MHC-I.
  JOURNAL   Nature. 2020 May;581(7806):100-105. doi: 10.1038/s41586-020-2229-5. 
            Epub 2020 Apr 22.
  PUBMED    32376951
REFERENCE   2  (bases 1 to 14709)
  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/s41586-020-2229-5"; journalName: "Nature"; date: "2020-05";
            volume: "581"; issue: "7806"; pages: "100-105"
COMMENT     Sequence Label: pSLIK_Hyg_mTurquoise2_Atg4BC74A
FEATURES             Location/Qualifiers
     source          1..14709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(132..151)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        323..702
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        704..902
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             920..1100
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1147..1272
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1765..1998
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2183..2227
                     /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    2525..2642
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    2699..2723
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    join(2773..2791,2792..2814,2815..2833,2834..2856,2857..2875,
                     2876..2898,2899..2917,2918..2940,2941..2959,2960..2982,
                     2983..3001,3002..3024,3025..3043)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        3076..3114
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3111..3135
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(3247..3249,3250..3252,3253..3963)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3292..3313)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3553..3572)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3900..3921
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5156..5180)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        5230..6441
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6278..6297
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(6566..7183,7184..7189,7190..7225,7226..7228,7229..7264,
                     7265..7267,7268..7303,7304..7312)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    7365..7938
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7532..7549)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7759..7778
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7997..9016
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             7997..9016
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     misc_feature    9482..10070
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9535..9555)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9953..9964)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10073..10089)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10074..10090)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10599..10779
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10805..10822)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10811..11035
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11081..11509
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11168..11187)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11378..11399
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11518..11538)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11523..11852
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11703..11838
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11765..11784
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11900..11947
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11966..12340
                     /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"
     polyA_signal    12470..12591
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12507..12526)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12561..12580
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12640..12656)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12640..12656)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12653..12675)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12664..12680
                     /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(12688..12694,12695..12712,12713..12718))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12733..12754
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12871..12888)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13042..13630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13122..13141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13801..14592,14593..14661))
                     /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     14424..14443
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(14662..14709,1..57))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
       61 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
      121 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
      181 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
      241 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      301 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      361 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      421 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      481 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      541 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      601 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      661 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      721 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      781 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      841 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      901 agcgcgtttt gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
      961 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     1021 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     1081 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
     1141 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
     1201 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
     1261 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
     1321 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     1381 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     1441 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     1501 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
     1561 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat
     1621 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     1681 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     1741 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     1801 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     1861 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     1921 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     1981 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     2041 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     2101 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     2161 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     2221 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     2281 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     2341 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     2401 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
     2461 attagtgaac ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc
     2521 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     2581 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     2641 ttcgggttta ttacagggac agcagagatc cagtttggtt aattctagac agttatcaac
     2701 aagtttgtac aaaaaagcag gcttcattac agggacagca gagatccagt ttggactagg
     2761 atcctttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta
     2821 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt
     2881 gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac
     2941 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
     3001 aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagct
     3061 cggtacccgg gtcgaggtag gcgtgtacgg tgggaggcct atataagcag agctcgttta
     3121 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
     3181 cgggaccgat ccagcctccg cggccccgaa ctagtccagt gtggtggaat tctgcagata
     3241 tcaaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     3301 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     3361 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     3421 cccaccctcg tgaccaccct gtcctggggc gtgcagtgct tcgcccgcta ccccgaccac
     3481 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     3541 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     3601 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     3661 gggcacaagc tggagtacaa ctactttagc gacaacgtct atatcaccgc cgacaagcag
     3721 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     3781 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3841 aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
     3901 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     3961 aagagatcca tggacgcagc cactttgaca tatgatactc tccggtttgc tgaatttgaa
     4021 gatttccctg agacctcaga gcctgtttgg attctgggca gaaaatacag cattttcaca
     4081 gagaaggacg aaatcttgtc tgatgttgca tccagacttt ggtttacata caggagaaac
     4141 tttccagcta ttgggggaac tggccctact tcagacacag gctggggtgc aatgcttcgg
     4201 tgtggacaga tgatctttgc ccaggccctg gtatgccggc acttaggtcg agattggagg
     4261 tggactcagc ggaagaggca gcctgacagc tactttaatg tcctcaatgc tttcctcgac
     4321 aggaaggaca gctactattc catccatcag atagcgcaaa tgggagttgg cgaaggcaag
     4381 tctataggcc agtggtacgg gccgaacact gttgcccagg tcctcaagaa acttgctgta
     4441 ttcgacacat ggagttcctt ggctgttcac atagcaatgg acaacactgt ggtgatggag
     4501 gaaatcagaa ggttatgcag ggccaacctt ccctgtgttg gggcggctgc acttcctact
     4561 gattcagaga ggcactgtaa tgggttccct gctggagctg aagtcaccaa caggccatcg
     4621 gcttggagac cactagtgct tctcatccct ctccgcctgg gactgacaga catcaatgag
     4681 gcctatgtgg agacgctgaa gcactgtttc atgatgcccc agtccctggg tgttattgga
     4741 gggaagccca acagtgccca ctactttatt ggctatgttg gggaggagct catctatctg
     4801 gacccccaca ctacacagcc tgcagtggag ctgactgaca gctgcttcat ccccgacgag
     4861 agcttccact gccagcaccc tccctctagg atgggcattg gagagcttga cccgtccatt
     4921 gctgtggggt ttttctgtaa gacggaggaa gactttaacg actggtgtca gcaagtcaaa
     4981 aagctatccc agcttggagg tgccctgccc atgtttgagc tggtggaaca gcagccttcc
     5041 catctggcct gccaggatgt cctgaacctg tccctagact cttctgatgt agagagactg
     5101 gagaggtttt ttgactctga ggatgaagac tttgaaatct tatccctctg ataaaaccca
     5161 gctttcttgt acaaagtggt tgataacatc tcgagccata aagatggtta attaacccac
     5221 ccaagatctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcctcacgg
     5281 cgagcgctgc cacgtcagac gaagggcgca gcgagcgtcc tgatccttcc gcccggacgc
     5341 tcaggacagc ggcccgctgc tcataagact cggccttaga accccagtat cagcagaagg
     5401 acattttagg acgggacttg ggtgactcta gggcactggt tttctttcca gagagcggaa
     5461 caggcgagga aaagtagtcc cttctcggcg attctgcgga gggatctccg tggggcggtg
     5521 aacgccgatg attatataag gacgcgccgg gtgtggcaca gctagttccg tcgcagccgg
     5581 gatttgggtc gcggttcttg tttgtggatc gctgtgatcg tcacttggtg agtagcgggc
     5641 tgctgggctg gccggggctt tcgtggccgc cgggccgctc ggtgggacgg aagcgtgtgg
     5701 agagaccgcc aagggctgta gtctgggtcc gcgagcaagg ttgccctgaa ctgggggttg
     5761 gggggagcgc agcaaaatgg cggctgttcc cgagtcttga atggaagacg cttgtgaggc
     5821 gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg gcggcaagaa cccaaggtct
     5881 tgaggccttc gctaatgcgg gaaagctctt attcgggtga gatgggctgg ggcaccatct
     5941 ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg
     6001 ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag cgcgcgccct
     6061 cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg ccacctgccg
     6121 gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct agggtaggct
     6181 ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag gcgtcagttt
     6241 ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt ttgaactatg
     6301 cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga aatgtaatca
     6361 tttgggtcaa tatgtaattt tcagtgttag actagtaaat tgtccgctaa attctggccg
     6421 tttttggctt ttttgttaga cgaagcttgg taccgagctc ggatctccac cccgtaccgg
     6481 tcctgcagga tttaaatggc gcgccgctag cgctaccgga ctcagatctc gagctcaagc
     6541 ttcgaattct gcagtcgaat tcaccatgtc tagactggac aagagcaaag tcataaacgg
     6601 agctctggaa ttactcaatg gtgtcggtat cgaaggcctg acgacaagga aactcgctca
     6661 aaagctggga gttgagcagc ctaccctgta ctggcacgtg aagaacaagc gggccctgct
     6721 cgatgccctg ccaatcgaga tgctggacag gcatcatacc cacttctgcc ccctggaagg
     6781 cgagtcatgg caagactttc tgcggaacaa cgccaagtca taccgctgtg ctctcctctc
     6841 acatcgcgac ggggctaaag tgcatctcgg cacccgccca acagagaaac agtacgaaac
     6901 cctggaaaat cagctcgcgt tcctgtgtca gcaaggcttc tccctggaga acgcactgta
     6961 cgctctgtcc gccgtgggcc actttacact gggctgcgta ttggaggaac aggagcatca
     7021 agtagcaaaa gaggaaagag agacacctac caccgattct atgcccccac ttctgagaca
     7081 agcaattgag ctgttcgacc ggcagggagc cgaacctgcc ttccttttcg gcctggaact
     7141 aatcatatgt ggcctggaga aacagctaaa gtgcgaaagc ggcgggccga ccgacgccct
     7201 tgacgatttt gacttagaca tgctcccagc cgatgccctt gacgactttg accttgatat
     7261 gctgcctgct gacgctcttg acgattttga ccttgacatg ctccccgggt aactaagtaa
     7321 ggatccgcgg ccgcactaga ggaattccgc ccctctcccc cccccccctc tccctccccc
     7381 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     7441 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     7501 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
     7561 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     7621 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     7681 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     7741 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     7801 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt
     7861 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     7921 tgaaaaacac gatgataagc ttaccggtac gcgtgatagc ttgccacaac ccgtaccaaa
     7981 gatggataga tccggaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga
     8041 aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt
     8101 cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt
     8161 ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt
     8221 gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg
     8281 tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga
     8341 ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg
     8401 accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc
     8461 ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc
     8521 tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc
     8581 ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg
     8641 gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc
     8701 gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc
     8761 aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag
     8821 cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt
     8881 ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg
     8941 gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc
     9001 gagggcaaag gaatagagta gatgccgacc gaacaagagc tgatttcgag aacgcctcag
     9061 ccagcaactc gcgcgagcct agcaaggcaa atgcgagaga acggccttac gcttggtggc
     9121 acagttctcg tccacagttc gctaagctcg ctcggctggg tcgcgggagg gccggtcgca
     9181 gtgattcagg cccttctgga ttgtgttggt ccccagggca cgattgtcat gcccacgcac
     9241 tcgggtgatc tgactgatcc cgcagattgg agatcgccgc ccgtgcctgc cgattgggtg
     9301 cagatctcta gatctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc
     9361 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     9421 cataaaatga atgcaattgt tgttgtttag tccctcccaa ttcgatatca agcttatcga
     9481 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     9541 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     9601 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     9661 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     9721 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     9781 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     9841 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     9901 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     9961 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
    10021 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
    10081 tcgacctcga tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta
    10141 ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca
    10201 cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
    10261 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg
    10321 atctgtggat ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc
    10381 cagggatcag atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag
    10441 agaaggtaga agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc
    10501 atgggatgga tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat
    10561 ttcatcacat ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga
    10621 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
    10681 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
    10741 ccctcagacc cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga
    10801 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
    10861 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
    10921 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
    10981 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
    11041 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
    11101 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
    11161 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
    11221 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
    11281 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
    11341 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
    11401 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
    11461 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
    11521 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
    11581 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
    11641 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
    11701 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
    11761 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
    11821 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
    11881 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
    11941 atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac
    12001 cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga
    12061 cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc
    12121 ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga
    12181 cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc
    12241 ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc
    12301 cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga
    12361 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
    12421 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
    12481 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
    12541 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
    12601 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
    12661 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
    12721 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
    12781 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
    12841 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
    12901 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
    12961 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
    13021 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
    13081 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
    13141 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
    13201 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
    13261 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
    13321 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
    13381 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
    13441 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
    13501 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
    13561 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
    13621 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
    13681 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
    13741 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
    13801 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
    13861 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
    13921 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
    13981 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
    14041 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
    14101 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
    14161 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
    14221 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
    14281 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
    14341 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
    14401 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
    14461 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
    14521 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
    14581 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
    14641 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
    14701 ttattgtct
//
