LOCUS       sequence-463086-       15448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression vector for an inducible sg09 resistant 
            PCSK5_M452I mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15448)
  AUTHORS   Marohl T, Atkins KA, Wang L, Janes KA
  TITLE     PCSK5M452I is a recessive hypomorph exclusive to MCF10DCIS.com 
            cells.
  JOURNAL   Mol Cancer Res. 2025 Oct 28. doi: 10.1158/1541-7786.MCR-25-0211.
  PUBMED    41148168
REFERENCE   2  (bases 1 to 15448)
  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: "Mol Cancer 
            Res. 2025 Oct 28. doi: 10.1158/1541-7786.MCR-25-0211."
COMMENT     Sequence Label: pSLIK PCSK5(sgRR,M452I)-V5 hygro
FEATURES             Location/Qualifiers
     source          1..15448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    2614..2638
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        2670..2984
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    2678..2696
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2714..2732
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2749..2767
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2785..2803
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2821..2839
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2856..2874
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2892..2910
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2957..2981
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5730..5771
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    complement(5810..5834)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        5884..7095
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6932..6951
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(7220..7837,7838..7843,7844..7879,7880..7882,7883..7918,
                     7919..7921,7922..7957,7958..7966)
                     /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    8019..8592
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8186..8203)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8413..8432
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8651..9670
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             8651..9670
                     /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    10136..10724
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10189..10209)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10607..10618)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10727..10743)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10728..10744)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             11253..11433
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11459..11476)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11465..11689
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11735..12163
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11822..11841)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12032..12053
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12172..12192)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12177..12506
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12357..12492
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12419..12438
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12554..12601
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12620..12994
                     /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    13124..13245
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13161..13180)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13215..13234
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13294..13310)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13294..13310)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13307..13329)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13318..13334
                     /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(13342..13348,13349..13366,13367..13372))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13387..13408
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13525..13542)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13696..14284)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13776..13795)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14455..15246,15247..15315))
                     /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     15078..15097
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15316..15420)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaattc tagacagtta tcaacaagtt tgtacaaaaa agcaggcttt
     2641 aaaggaacca attcagtcga ctggatcccg agtttactcc ctatcagtga tagagaacgt
     2701 atgtcgagtt tactccctat cagtgataga gaacgatgtc gagtttactc cctatcagtg
     2761 atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttac
     2821 tccctatcag tgatagagaa cgtatgtcga gtttatccct atcagtgata gagaacgtat
     2881 gtcgagttta ctccctatca gtgatagaga acgtatgtcg aggtaggcgt gtacggtggg
     2941 aggcctatat aagcagagct cgtttagtga accgtcagat cgccactagt atgggctggg
     3001 ggagccgctg ctgctgcccg ggacgtttgg acctgctgtg cgtgctggcg ctgctcgggg
     3061 gctgcctgct ccccgtgtgt cggacgcgcg tctacaccaa ccactgggca gtcaaaatcg
     3121 ccgggggctt cccggaggcc aaccgtatcg ccagcaagta cggattcatc aacataggac
     3181 agataggggc cctgaaggac tactaccact tctaccatag caggacgatt aaaaggtcag
     3241 ttatctcgag cagagggacc cacagtttca tttcaatgga accaaaggtg gaatggatcc
     3301 aacagcaagt ggtaaaaaag cggacaaaga gggattatga cttcagtcgt gcccagtcta
     3361 cctatttcaa tgatcccaag tggcccagca tgtggtatat gcactgcagt gacaatacac
     3421 atccctgcca gtctgacatg aatatcgaag gagcctggaa gagaggctac acggggaaga
     3481 acatcgtcgt cactatcctg gatgacggaa ttgagagaac ccatccagat ctgatgcaaa
     3541 actacgatgc tctggcaagt tgcgacgtga atgggaatga cttggaccca atgcctcgtt
     3601 atgatgcaag caacgagaac aagcatggga ctcgctgtgc tggagaagtg gcagccgctg
     3661 caaacaattc gcactgcaca gtcggaattg ctttcaacgc caagatcgga ggagtgcgaa
     3721 tgctggacgg agatgtcacg gacatggttg aagcaaaatc agttagcttc aacccccagc
     3781 acgtgcacat ttacagcgcc agctggggcc cggatgatga tggcaagact gtggacggac
     3841 cagcccccct cacccggcaa gcctttgaaa acggcgttag aatggggcgg agaggcctcg
     3901 gctctgtgtt tgtttgggca tctggaaatg gtggaaggag caaagaccac tgctcctgtg
     3961 atggctacac caacagcatc tacaccatct ccatcagcag cactgcagaa agcggaaaga
     4021 aaccttggta cctggaagag tgttcatcca cgctggccac aacctacagc agcggggagt
     4081 cctacgataa gaaaatcatc actacagatc tgaggcagcg ttgcacggac aaccacactg
     4141 ggacgtcagc ctcagccccc atggctgcag gcatcattgc gctggccctg gaagccaatc
     4201 cgtttctgac ctggagagac gtacagcatg ttattgtcag gacttcccgt gcgggacatt
     4261 tgaacgctaa tgactggaaa accaatgctg ctggttttaa ggtgagccat ctttatggat
     4321 ttggactgat ggacgcagaa gccattgtga tggaggcaga gaagtggacc accgttcccc
     4381 ggcagcacgt gtgtgtggag agcacagacc gacaaatcaa gacaatccgc cctaacagtg
     4441 cagtgcgctc catctacaaa gcttcaggct gctcggataa ccccaaccgc catgtcaact
     4501 acctggagca cgtcgttgtg cgcatcacca tcacccaccc caggagagga gacctggcca
     4561 tctacctgac ctcgccctct ggaactaggt ctcagctttt ggccaacagg ctatttgatc
     4621 actccatgga aggattcaaa aactgggagt tcatgaccat tcattgctgg ggagaaagag
     4681 ctgctggtga ctgggtcctt gaagtttatg atactccctc tcagctaagg aactttaaga
     4741 ctccaggtaa attgaaagaa tggtctttgg tcctctacgg cacctccgtg cagccatatt
     4801 caccaaccaa tgaatttccg aaagtggaac ggttccgcta tagccgagtt gaagacccca
     4861 cagacgacta tggcacagag gattatgcag gtccctgcga ccctgagtgc agtgaggttg
     4921 gctgtgacgg gccaggacca gaccactgca atgactgttt gcactactac tacaagctga
     4981 aaaacaatac caggatctgt gtctccagct gcccccctgg ccactaccac gccgacaaga
     5041 agcgctgcag gaagtgtgcc cccaactgtg agtcctgctt tgggagccat ggtgaccaat
     5101 gcatgtcctg caaatatgga tactttctga atgaagaaac caacagctgt gttactcact
     5161 gccctgatgg gtcatatcag gataccaaga aaaatctttg ccggaaatgc agtgaaaact
     5221 gcaagacatg tactgaattc cataactgta cagaatgtag ggatgggtta agcctgcagg
     5281 gatcccggtg ctctgtctcc tgtgaagatg gacggtattt caacggccag gactgccagc
     5341 cctgccaccg cttctgcgcc acttgtgctg gggcaggagc tgatgggtgc attaactgca
     5401 cagagggcta cttcatggag gatgggagat gcgtgcagag ctgtagtatc agctattact
     5461 ttgaccactc ttcagagaat ggatacaaat cctgcaaaaa atgtgatatc agttgtttga
     5521 cgtgcaatgg cccaggattc aagaactgta caagctgccc tagtgggtat ctcttagact
     5581 taggaatgtg tcaaatggga gccatttgca aggatgcaac ggaagagtcc tgggcggaag
     5641 gaggcttctg tatgcttgtg aaaaagaaca atctgtgcca acggaaggtt cttcaacaac
     5701 tttgctgcaa aacatgtaca tttcaaggcg gtaagcctat ccctaaccct ctcctcggtc
     5761 tcgattctac gtaacaattg gaagactaat gcgttttcga gatatctaga cccagctttc
     5821 ttgtacaaag tggttgataa catctcgagc cataaagatg gttaattaac ccacccaaga
     5881 tctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcctc acggcgagcg
     5941 ctgccacgtc agacgaaggg cgcagcgagc gtcctgatcc ttccgcccgg acgctcagga
     6001 cagcggcccg ctgctcataa gactcggcct tagaacccca gtatcagcag aaggacattt
     6061 taggacggga cttgggtgac tctagggcac tggttttctt tccagagagc ggaacaggcg
     6121 aggaaaagta gtcccttctc ggcgattctg cggagggatc tccgtggggc ggtgaacgcc
     6181 gatgattata taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg
     6241 ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg
     6301 gctggccggg gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt gtggagagac
     6361 cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg gttgggggga
     6421 gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg
     6481 tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag gtcttgaggc
     6541 cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctggggcacc atctggggac
     6601 cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg cgggggcggc
     6661 agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt
     6721 gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacct gccggtaggt
     6781 gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta ggctctcctg
     6841 aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca gtttctttgg
     6901 tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg
     6961 gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta atcatttggg
     7021 tcaatatgta attttcagtg ttagactagt aaattgtccg ctaaattctg gccgtttttg
     7081 gcttttttgt tagacgaagc ttggtaccga gctcggatct ccaccccgta ccggtcctgc
     7141 aggatttaaa tggcgcgccg ctagcgctac cggactcaga tctcgagctc aagcttcgaa
     7201 ttctgcagtc gaattcacca tgtctagact ggacaagagc aaagtcataa acggagctct
     7261 ggaattactc aatggtgtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct
     7321 gggagttgag cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc
     7381 cctgccaatc gagatgctgg acaggcatca tacccacttc tgccccctgg aaggcgagtc
     7441 atggcaagac tttctgcgga acaacgccaa gtcataccgc tgtgctctcc tctcacatcg
     7501 cgacggggct aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga
     7561 aaatcagctc gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct
     7621 gtccgccgtg ggccacttta cactgggctg cgtattggag gaacaggagc atcaagtagc
     7681 aaaagaggaa agagagacac ctaccaccga ttctatgccc ccacttctga gacaagcaat
     7741 tgagctgttc gaccggcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat
     7801 atgtggcctg gagaaacagc taaagtgcga aagcggcggg ccgaccgacg cccttgacga
     7861 ttttgactta gacatgctcc cagccgatgc ccttgacgac tttgaccttg atatgctgcc
     7921 tgctgacgct cttgacgatt ttgaccttga catgctcccc gggtaactaa gtaaggatcc
     7981 gcggccgcac tagaggaatt ccgcccctct cccccccccc cctctccctc ccccccccct
     8041 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     8101 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     8161 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     8221 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     8281 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     8341 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     8401 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     8461 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     8521 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     8581 acacgatgat aagcttaccg gtacgcgtga tagcttgcca caacccgtac caaagatgga
     8641 tagatccgga aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
     8701 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
     8761 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
     8821 agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga
     8881 cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     8941 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat
     9001 ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
     9061 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
     9121 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
     9181 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
     9241 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
     9301 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
     9361 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
     9421 gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     9481 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     9541 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga
     9601 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
     9661 aaaggaatag agtagatgcc gaccgaacaa gagctgattt cgagaacgcc tcagccagca
     9721 actcgcgcga gcctagcaag gcaaatgcga gagaacggcc ttacgcttgg tggcacagtt
     9781 ctcgtccaca gttcgctaag ctcgctcggc tgggtcgcgg gagggccggt cgcagtgatt
     9841 caggcccttc tggattgtgt tggtccccag ggcacgattg tcatgcccac gcactcgggt
     9901 gatctgactg atcccgcaga ttggagatcg ccgcccgtgc ctgccgattg ggtgcagatc
     9961 tctagatctg tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt
    10021 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
    10081 atgaatgcaa ttgttgttgt ttagtccctc ccaattcgat atcaagctta tcgataatca
    10141 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
    10201 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
    10261 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
    10321 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
    10381 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
    10441 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
    10501 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
    10561 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
    10621 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
    10681 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc
    10741 tcgatcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg
    10801 ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc
    10861 aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
    10921 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt
    10981 ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca gggccaggga
    11041 tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag caagagaagg
    11101 tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc ctgcatggga
    11161 tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta gcatttcatc
    11221 acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac cagatctgag
    11281 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
    11341 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
    11401 gaccctttta gtcagtgtgg aaaatctcta gcagggcccg tttaaacccg ctgatcagcc
    11461 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
    11521 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
    11581 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
    11641 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
    11701 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
    11761 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
    11821 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
    11881 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
    11941 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
    12001 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
    12061 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
    12121 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
    12181 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
    12241 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
    12301 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
    12361 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
    12421 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
    12481 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
    12541 ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
    12601 caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg
    12661 cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt
    12721 ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca
    12781 ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct
    12841 gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat
    12901 gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa
    12961 ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt tcgattccac
    13021 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
    13081 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
    13141 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
    13201 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
    13261 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
    13321 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
    13381 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
    13441 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
    13501 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
    13561 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
    13621 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
    13681 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
    13741 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
    13801 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
    13861 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
    13921 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
    13981 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
    14041 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
    14101 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
    14161 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
    14221 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
    14281 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
    14341 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
    14401 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
    14461 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
    14521 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
    14581 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
    14641 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
    14701 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
    14761 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
    14821 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
    14881 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
    14941 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
    15001 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
    15061 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
    15121 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
    15181 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
    15241 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
    15301 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
    15361 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
    15421 cacatttccc cgaaaagtgc cacctgac
//
