LOCUS       sequence-454818-       13090 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of Gaussia luciferase and shRNAmir of interest.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13090)
  AUTHORS   Charles JP, Fuchs J, Hefter M, Vischedyk JB, Kleint M, Vogiatzi F, 
            Schafer JA, Nist A, Timofeev O, Wanzel M, Stiewe T
  TITLE     Monitoring the dynamics of clonal tumour evolution in vivo using 
            secreted luciferases.
  JOURNAL   Nat Commun. 2014 Jun 3;5:3981. doi: 10.1038/ncomms4981.
  PUBMED    24889111
REFERENCE   2  (bases 1 to 13090)
  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: "2014-06-3"; volume: "5:3981. doi"; pages: " 
            10.1038/ncomms4981"
COMMENT     Sequence Label: pIND-GLucZ
FEATURES             Location/Qualifiers
     source          1..13090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    462..695
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             880..924
                     /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    1222..1339
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(1404..1778)
                     /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      1775..1784
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1788..1806)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1798..1845)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    join(1915..1933,1934..1956,1957..1975,1976..1998,1999..2017,
                     2018..2040,2041..2059,2060..2082,2083..2101,2102..2124,
                     2125..2143,2144..2166,2167..2185)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2218..2256
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2253..2277
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2372..2381
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2378..2935
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     ncRNA           2985..3106
                     /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           3134..3148
                     /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           3176..3302
                     /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"
     promoter        3353..4564
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4401..4420
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    4571..4604
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             join(4611..5228,5229..5234,5235..5270,5271..5273,5274..5309,
                     5310..5312,5313..5348,5349..5357)
                     /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    5406..5978
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5573..5590)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5800..5819
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5981..6583
                     /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     6480..6500
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6605..7193
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6658..6678)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7076..7087)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7400..7633
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7734..7751)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7740..7964
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8010..8438
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8097..8116)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8307..8328
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8447..8467)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8452..8781
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8632..8767
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8694..8713
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    9604..9725
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9774..9790)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9774..9790)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9787..9809)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9798..9814
                     /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(9822..9828,9829..9846,9847..9852))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9867..9888
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10005..10022)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10176..10764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10256..10275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10935..11726,11727..11795))
                     /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     11558..11577
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11796..11900)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    11948..12082
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             12250..12883
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    12930..13055
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
ORIGIN
        1 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
       61 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
      121 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
      181 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
      241 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
      301 agcggccggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
      361 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
      421 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
      481 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
      541 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
      601 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
      661 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
      721 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
      781 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
      841 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
      901 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
      961 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1021 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1081 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1141 agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca
     1201 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     1261 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     1321 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggttagt
     1381 accgggcccg ctctagtccg gaatcagtcc tgctcctcgg ccacgaagtg cacgcagttg
     1441 ccggccgggt cgcgcagggc gaactcccgc ccccacggct gctcgccgat ctcggtcatg
     1501 gccggcccgg aggcgtcccg gaagttcgtg gacacgacct ccgaccactc ggcgtacagc
     1561 tcgtccaggc cgcgcaccca cacccaggcc agggtgttgt ccggcaccac ctggtcctgg
     1621 accgcgctga tgaacagggt cacgtcgtcc cggaccacac cggcgaagtc gtcctccacg
     1681 aagtcccggg agaacccgag ccggtcggtc cagaactcga ccgctccggc gacgtcgcgc
     1741 gcggtgagca ccggaacggc actggtcaac ttggccatgg tggccctcct atagtgagtc
     1801 gtattatact atgccgatat actatgccga tgattaattg tcaacacgtg ctgcaggtcc
     1861 gaggttctag gctagcttaa aggaaccaat tcagtcgact ggatccttta ccactcccta
     1921 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt
     1981 gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac
     2041 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
     2101 aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt
     2161 taccactccc tatcagtgat agagaaaagt gaaagtcgag ctcggtaccc gggtcgaggt
     2221 aggcgtgtac ggtgggaggc ctatataagc agagctcgtt tagtgaaccg tcagatcgcc
     2281 tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg atccagccta
     2341 ccggtcaagc ttggtaccga gctcggatcc agccaccatg ggagtcaaag ttctgtttgc
     2401 cctgatctgc atcgctgtgg ccgaggccaa gcccaccgag aacaacgaag acttcaacat
     2461 cgtggccgtg gccagcaact tcgcgaccac ggatctcgat gctgaccgcg ggaagttgcc
     2521 cggcaagaag ctgccgctgg aggtgctcaa agagatggaa gccaatgccc ggaaagctgg
     2581 ctgcaccagg ggctgtctga tctgcctgtc ccacatcaag tgcacgccca agatgaagaa
     2641 gttcatccca ggacgctgcc acacctacga aggcgacaaa gagtccgcac agggcggcat
     2701 aggcgaggcg atcgtcgaca ttcctgagat tcctgggttc aaggacttgg agcccatgga
     2761 gcagttcatc gcacaggtcg atctgtgtgt ggactgcaca actggctgcc tcaaagggct
     2821 tgccaacgtg cagtgttctg acctgctcaa gaagtggctg ccgcaacgct gtgcgacctt
     2881 tgccagcaag atccagggcc aggtggacaa gatcaagggg gccggtggtg actaagcggc
     2941 cgcaagcctt gttaagtgct cgcttcggca gcacatatac tatgttgaat gaggcttcag
     3001 tactttacag aatcgttgcc tgcacatctt ggaaacactt gctgggatta cttcttcagg
     3061 ttaacccaac agaaggctcg agaaggtata ttgctgttga cagtgagcga tctcgcttgg
     3121 gcgagagtaa gtagtgaagc cacagatgta cttactctcg cccaagcgag agtgcctact
     3181 gcctcggaat tcaaggggct actttaggag caattatctt gtttactaaa actgaatacc
     3241 ttgctatctc tttgatacat ttttacaaag ctgaattaaa atggtataaa ttaaatcact
     3301 tttttcaatt ggaagactaa tgcgtttaaa cacgcggcga cgcgtaagat ctggcctccg
     3361 cgccgggttt tggcgcctcc cgcgggcgcc cccctcctca cggcgagcgc tgccacgtca
     3421 gacgaagggc gcagcgagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc
     3481 tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac
     3541 ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag
     3601 tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat
     3661 aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc
     3721 ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg
     3781 ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct
     3841 gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa
     3901 tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg
     3961 aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgaggcc ttcgctaatg
     4021 cgggaaagct cttattcggg tgagatgggc tggggcacca tctggggacc ctgacgtgaa
     4081 gtttgtcact gactggagaa ctcggtttgt cgtctgttgc gggggcggca gttatggcgg
     4141 tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc
     4201 acccgttctg ttggcttata atgcagggtg gggccacctg ccggtaggtg tgtggtaggc
     4261 ttttctccgt cgcaggacgc agggttcggg cctagggtag gctctcctga atcgacaggc
     4321 gccggacctc tggtgagggg agggataagt gaggcgtcag tttctttggt cggttttatg
     4381 tacctatctt cttaagtagc tgaagctccg gttttgaact atgcgctcgg ggttggcgag
     4441 tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa
     4501 ttttcagtgt tagactagta aattgtccgc taaattctgg ccgtttttgg cttttttgtt
     4561 agacggatcc ataacttcgt ataatgtatg ctatacgaag ttatgccacc atgtctagac
     4621 tggacaagag caaagtcata aacggagctc tggaattact caatggtgtc ggtatcgaag
     4681 gcctgacgac aaggaaactc gctcaaaagc tgggagttga gcagcctacc ctgtactggc
     4741 acgtgaagaa caagcgggcc ctgctcgatg ccctgccaat cgagatgctg gacaggcatc
     4801 atacccactt ctgccccctg gaaggcgagt catggcaaga ctttctgcgg aacaacgcca
     4861 agtcataccg ctgtgctctc ctctcacatc gcgacggggc taaagtgcat ctcggcaccc
     4921 gcccaacaga gaaacagtac gaaaccctgg aaaatcagct cgcgttcctg tgtcagcaag
     4981 gcttctccct ggagaacgca ctgtacgctc tgtccgccgt gggccacttt acactgggct
     5041 gcgtattgga ggaacaggag catcaagtag caaaagagga aagagagaca cctaccaccg
     5101 attctatgcc cccacttctg agacaagcaa ttgagctgtt cgaccggcag ggagccgaac
     5161 ctgccttcct tttcggcctg gaactaatca tatgtggcct ggagaaacag ctaaagtgcg
     5221 aaagcggcgg gccgaccgac gcccttgacg attttgactt agacatgctc ccagccgatg
     5281 cccttgacga ctttgacctt gatatgctgc ctgctgacgc tcttgacgat tttgaccttg
     5341 acatgctccc cgggtaaata acttcgtata gcatacatta tacgaagtat ggcgcgccaa
     5401 ttccgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     5461 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     5521 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     5581 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     5641 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     5701 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     5761 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     5821 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     5881 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     5941 cacggggacg tggttttcct ttgaaaaaca cgataatacc atggccaccg agtacaagcc
     6001 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc
     6061 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt
     6121 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     6181 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     6241 ggtgttcgcc gagatcggct cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     6301 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     6361 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     6421 cggagtggag gcggccgagc gcgctggggt gcccgccttc ctggagacct ccgcgccccg
     6481 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     6541 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgattaatta agcgcgtctg
     6601 gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     6661 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     6721 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     6781 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     6841 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     6901 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     6961 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     7021 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     7081 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     7141 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat
     7201 taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct
     7261 accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc
     7321 acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     7381 ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca agatctgctt
     7441 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     7501 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     7561 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     7621 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     7681 aaatgaatat cagagagtga gaggccttga cattgtttaa acccgctgat cagcctcgac
     7741 tgtgccttct agttgccagc catctgttgt ttgccccttc cccgtgcctt ccttgaccct
     7801 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     7861 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     7921 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     7981 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     8041 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     8101 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     8161 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     8221 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     8281 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     8341 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     8401 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     8461 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     8521 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     8581 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     8641 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     8701 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     8761 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     8821 cagcacgtga tgaaaaagcc tgaactcacc gcgacgtgac cgcaaggaat cggtcaatac
     8881 actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact
     8941 gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg
     9001 gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc
     9061 ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat
     9121 tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag
     9181 cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg
     9241 tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat
     9301 gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc
     9361 gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta
     9421 ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga atagcacgtg
     9481 ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     9541 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     9601 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     9661 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact aatcaatgta
     9721 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     9781 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     9841 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     9901 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     9961 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
    10021 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
    10081 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
    10141 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
    10201 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
    10261 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
    10321 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
    10381 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcactaaccc
    10441 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
    10501 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
    10561 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
    10621 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
    10681 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
    10741 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
    10801 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
    10861 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
    10921 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
    10981 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
    11041 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
    11101 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
    11161 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
    11221 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
    11281 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
    11341 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
    11401 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
    11461 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
    11521 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
    11581 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
    11641 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
    11701 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
    11761 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
    11821 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
    11881 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
    11941 ggagatcaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
    12001 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
    12061 tgtatcttat catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact
    12121 tcccacccca taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg
    12181 tgattcctct gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa
    12241 cttagtagtt ggaagggcta attcactccc aaagaagaca agatatcctt gatctgtgga
    12301 tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca
    12361 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag
    12421 aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg
    12481 atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg
    12541 tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg
    12601 gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag
    12661 ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac
    12721 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
    12781 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
    12841 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
    12901 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
    12961 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
    13021 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat
    13081 cgcgatggga
//
