LOCUS       sequence-83326-e       14250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of Cypridina luciferase and shRNAmir of 
            interest.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14250)
  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 14250)
  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-CLucZ
FEATURES             Location/Qualifiers
     source          1..14250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    497..730
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             915..959
                     /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    1257..1374
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(1439..1813)
                     /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      1810..1819
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1823..1841)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1833..1880)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    join(1950..1968,1969..1991,1992..2010,2011..2033,2034..2052,
                     2053..2075,2076..2094,2095..2117,2118..2136,2137..2159,
                     2160..2178,2179..2201,2202..2220)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2253..2291
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2288..2312
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2412..4073
                     /codon_start=1
                     /product="secreted Cypridina luciferase"
                     /label=CLuc
                     /note="contains 30 codon substitutions for efficient 
                     translation in mammalian cells"
                     /translation="MKTLILAVALVYCATVHCQDCPYEPDPPNTVPTSCEAKEGECIDS
                     SCGTCTRDILSDGLCENKPGKTCCRMCQYVIECRVEAAGWFRTFYGKRFQFQEPGTYVL
                     GQGTKGGDWKVSITLENLDGTKGAVLTKTRLEVAGDIIDIAQATENPITVNGGADPIIA
                     NPYTIGEVTIAVVEMPGFNITVIEFFKLIVIDILGGRSVRIAPDTANKGMISGLCGDLK
                     MMEDTDFTSDPEQLAIQPKINQEFDGCPLYGNPDDVAYCKGLLEPYKDSCRNPINFYYY
                     TISCAFARCMGGDERASHVLLDYRETCAAPETRGTCVLSGHTFYDTFDKARYQFQGPCK
                     EILMAADCFWNTWDVKVSHRNVDSYTEVEKVRIRKQSTVVELIVDGKQILVGGEAVSVP
                     YSSQNTSIYWQDGDILTTAILPEALVVKFNFKQLLVVHIRDPFDGKTCGICGNYNQDFS
                     DDSFDAEGACDLTPNPPGCTEEQKPEAERLCNSLFAGQSDLDQKCNVCHKPDRVERCMY
                     EYCLRGQQGFCDHAWEFKKECYIKHGDTLEVPDECK"
     ncRNA           4123..4244
                     /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           4272..4286
                     /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           4314..4440
                     /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        4491..5702
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5539..5558
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    5709..5742
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             join(5749..6366,6367..6372,6373..6408,6409..6411,6412..6447,
                     6448..6450,6451..6486,6487..6495)
                     /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"
     protein_bind    complement(6496..6529)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6545..7117
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6712..6729)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6939..6958
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7120..7722
                     /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     7619..7639
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7744..8332
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7797..7817)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8215..8226)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8539..8772
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8873..8890)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8879..9103
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9149..9577
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9236..9255)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9446..9467
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9586..9606)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9591..9920
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9771..9906
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9833..9852
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    10743..10864
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10913..10929)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10913..10929)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10926..10948)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10937..10953
                     /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(10961..10967,10968..10985,10986..10991))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11006..11027
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11144..11161)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11315..11903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11395..11414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12074..12865,12866..12934))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRMDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     12697..12716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12935..13039)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    13087..13221
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             13389..14022
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 ccgagtaata ccgggggaga ttagatcgcg atgggaaaaa ttcggttaag gccaggggga
       61 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
      121 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
      181 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
      241 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
      301 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac
      361 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
      421 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
      481 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
      541 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
      601 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
      661 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
      721 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
      781 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
      841 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
      901 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
      961 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     1021 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     1081 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     1141 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     1201 acggatcggc actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt
     1261 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     1321 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
     1381 tattacaggg acagcagaga tccagtttgg ttagtaccgg gcccgctcta gtccggaatc
     1441 agtcctgctc ctcggccacg aagtgcacgc agttgccggc cgggtcgcgc agggcgaact
     1501 cccgccccca cggctgctcg ccgatctcgg tcatggccgg cccggaggcg tcccggaagt
     1561 tcgtggacac gacctccgac cactcggcgt acagctcgtc caggccgcgc acccacaccc
     1621 aggccagggt gttgtccggc accacctggt cctggaccgc gctgatgaac agggtcacgt
     1681 cgtcccggac cacaccggcg aagtcgtcct ccacgaagtc ccgggagaac ccgagccggt
     1741 cggtccagaa ctcgaccgct ccggcgacgt cgcgcgcggt gagcaccgga acggcactgg
     1801 tcaacttggc catggtggcc ctcctatagt gagtcgtatt atactatgcc gatatactat
     1861 gccgatgatt aattgtcaac acgtgctgca ggtccgaggt tctaggctag cttaaaggaa
     1921 ccaattcagt cgactggatc ctttaccact ccctatcagt gatagagaaa agtgaaagtc
     1981 gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat
     2041 cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg
     2101 aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact
     2161 ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga
     2221 aaagtgaaag tcgagctcgg tacccgggtc gaggtaggcg tgtacggtgg gaggcctata
     2281 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
     2341 acctccatag aagacaccgg gaccgatcca gcctaccggt caagcttggt accgagctcg
     2401 gatccgccac catgaagacc ttaattcttg ccgttgcatt agtctactgc gccactgttc
     2461 attgccagga ctgtccttac gaacctgatc caccaaacac agttccaact tcctgtgaag
     2521 ctaaagaagg agaatgtatt gatagcagct gtggcacctg cacgagagac atactatcag
     2581 atggactgtg tgaaaataaa ccaggaaaaa catgttgccg aatgtgtcag tatgtaattg
     2641 aatgcagagt agaggccgca ggatggttta gaacattcta tggaaagaga ttccagttcc
     2701 aggaacctgg tacatacgtg ttgggtcaag gaaccaaggg cggcgactgg aaggtgtcca
     2761 tcaccctgga gaacctggat ggaaccaagg gggctgtgct gaccaagaca agactggaag
     2821 tggctggaga catcattgac atcgctcaag ctactgagaa tcccatcact gtaaacggtg
     2881 gagctgaccc tatcatcgcc aacccgtaca ccatcggcga ggtcaccatc gctgttgttg
     2941 agatgccagg cttcaacatc accgtcattg agttcttcaa actgatcgtg atcgacatcc
     3001 tcggaggaag atctgtaaga atcgccccag acacagcaaa caaaggaatg atctctggcc
     3061 tctgtggaga tcttaaaatg atggaagata cagacttcac ttcagatcca gaacaactcg
     3121 ctattcagcc taagatcaac caggagtttg acggttgtcc actctatgga aatcctgatg
     3181 acgttgcata ctgcaaaggt cttctggagc cgtacaagga cagctgccgc aaccccatca
     3241 acttctacta ctacaccatc tcctgcgcct tcgcccgctg tatgggtgga gacgagcgag
     3301 cctcacacgt gctgcttgac tacagggaga cgtgcgctgc tcccgaaact agaggaacct
     3361 gcgttttgtc tggacatact ttctacgata catttgacaa agcaagatac caattccagg
     3421 gtccctgcaa ggagattctt atggccgccg actgtttctg gaacacttgg gatgtgaagg
     3481 tttcacacag gaatgttgac tcttacactg aagtagagaa agtacgaatc aggaaacaat
     3541 cgactgtagt agaactcatt gttgatggaa aacagattct ggttggagga gaagccgtgt
     3601 ccgtcccgta cagctctcag aacacttcca tctactggca agatggtgac atactgacta
     3661 cagccatcct acctgaagct ctggtggtca agttcaactt caagcaactg ctcgtcgtac
     3721 atattagaga tccattcgat ggtaagactt gcggtatttg cggtaactac aaccaggatt
     3781 tcagtgatga ttcttttgat gctgaaggag cctgtgatct gacccccaac ccaccgggat
     3841 gcaccgaaga acagaaacct gaagctgaac gactctgcaa tagtctcttc gccggtcaaa
     3901 gtgatcttga tcagaaatgt aacgtgtgcc acaagcctga ccgtgtcgaa cgatgcatgt
     3961 acgagtattg cctgagggga caacagggtt tctgtgacca cgcatgggag ttcaagaaag
     4021 aatgctacat aaagcatgga gacaccctag aagtaccaga tgaatgcaaa taggcggccg
     4081 caagccttgt taagtgctcg cttcggcagc acatatacta tgttgaatga ggcttcagta
     4141 ctttacagaa tcgttgcctg cacatcttgg aaacacttgc tgggattact tcttcaggtt
     4201 aacccaacag aaggctcgag aaggtatatt gctgttgaca gtgagcgatc tcgcttgggc
     4261 gagagtaagt agtgaagcca cagatgtact tactctcgcc caagcgagag tgcctactgc
     4321 ctcggaattc aaggggctac tttaggagca attatcttgt ttactaaaac tgaatacctt
     4381 gctatctctt tgatacattt ttacaaagct gaattaaaat ggtataaatt aaatcacttt
     4441 tttcaattgg aagactaatg cgtttaaaca cgcggcgacg cgtaagatct ggcctccgcg
     4501 ccgggttttg gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg ccacgtcaga
     4561 cgaagggcgc agcgagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg
     4621 ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt
     4681 gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc
     4741 ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa
     4801 ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt
     4861 gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct
     4921 ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt
     4981 agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg
     5041 gcggctgttc ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa
     5101 acaaggtggg gggcatggtg ggcggcaaga acccaaggtc ttgaggcctt cgctaatgcg
     5161 ggaaagctct tattcgggtg agatgggctg gggcaccatc tggggaccct gacgtgaagt
     5221 ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg
     5281 ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac
     5341 ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt
     5401 ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc
     5461 cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta
     5521 cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg
     5581 tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt
     5641 ttcagtgtta gactagtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag
     5701 acggatccat aacttcgtat aatgtatgct atacgaagtt atgccaccat gtctagactg
     5761 gacaagagca aagtcataaa cggagctctg gaattactca atggtgtcgg tatcgaaggc
     5821 ctgacgacaa ggaaactcgc tcaaaagctg ggagttgagc agcctaccct gtactggcac
     5881 gtgaagaaca agcgggccct gctcgatgcc ctgccaatcg agatgctgga caggcatcat
     5941 acccacttct gccccctgga aggcgagtca tggcaagact ttctgcggaa caacgccaag
     6001 tcataccgct gtgctctcct ctcacatcgc gacggggcta aagtgcatct cggcacccgc
     6061 ccaacagaga aacagtacga aaccctggaa aatcagctcg cgttcctgtg tcagcaaggc
     6121 ttctccctgg agaacgcact gtacgctctg tccgccgtgg gccactttac actgggctgc
     6181 gtattggagg aacaggagca tcaagtagca aaagaggaaa gagagacacc taccaccgat
     6241 tctatgcccc cacttctgag acaagcaatt gagctgttcg accggcaggg agccgaacct
     6301 gccttccttt tcggcctgga actaatcata tgtggcctgg agaaacagct aaagtgcgaa
     6361 agcggcgggc cgaccgacgc ccttgacgat tttgacttag acatgctccc agccgatgcc
     6421 cttgacgact ttgaccttga tatgctgcct gctgacgctc ttgacgattt tgaccttgac
     6481 atgctccccg ggtaaataac ttcgtatagc atacattata cgaagttatg gcgcgccaat
     6541 tccgcccctc tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc
     6601 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg
     6661 cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca
     6721 aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa
     6781 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt
     6841 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt
     6901 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca
     6961 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc
     7021 ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc
     7081 acggggacgt ggttttcctt tgaaaaacac gataatacca tggccaccga gtacaagccc
     7141 acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg
     7201 ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc
     7261 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     7321 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     7381 gtgttcgccg agatcggctc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     7441 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     7501 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     7561 ggagtggagg cggccgagcg cgctggggtg cccgccttcc tggagacctc cgcgccccgc
     7621 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     7681 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gattaattaa gcgcgtctgg
     7741 aacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     7801 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     7861 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     7921 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     7981 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     8041 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     8101 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     8161 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     8221 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     8281 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt
     8341 aattctgcag tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta
     8401 ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca
     8461 cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     8521 taaaagaaaa gaggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt
     8581 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     8641 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     8701 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     8761 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     8821 aatgaatatc agagagtgag aggccttgac attgtttaaa cccgctgatc agcctcgact
     8881 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     8941 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     9001 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     9061 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     9121 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     9181 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     9241 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     9301 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     9361 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     9421 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     9481 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     9541 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     9601 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     9661 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     9721 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     9781 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     9841 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     9901 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     9961 agcacgtgat gaaaaagcct gaactcaccg cgacgtgacc gcaaggaatc ggtcaataca
    10021 ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg
    10081 tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg
    10141 ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc
    10201 tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt
    10261 cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc
    10321 agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt
    10381 atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg
    10441 atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg
    10501 ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac
    10561 tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa tagcacgtgc
    10621 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
    10681 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
    10741 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
    10801 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
    10861 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
    10921 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
    10981 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
    11041 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
    11101 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
    11161 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
    11221 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
    11281 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
    11341 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
    11401 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
    11461 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
    11521 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
    11581 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
    11641 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
    11701 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
    11761 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
    11821 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
    11881 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
    11941 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
    12001 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
    12061 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
    12121 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
    12181 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
    12241 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
    12301 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
    12361 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
    12421 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
    12481 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
    12541 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
    12601 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
    12661 ggcgaccgag ttgctcttgc ccggcgtcca tacgggataa taccgcgcca catagcagaa
    12721 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
    12781 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
    12841 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
    12901 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
    12961 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
    13021 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
    13081 gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
    13141 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
    13201 gtatcttatc atgtctggat caactggata actcaagcta accaaaatca tcccaaactt
    13261 cccaccccat accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt
    13321 gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac
    13381 ttagtagttg gaagggctaa ttcactccca aagaagacaa gatatccttg atctgtggat
    13441 ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc caggggtcag
    13501 atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga
    13561 agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga
    13621 tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt
    13681 ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg
    13741 actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc
    13801 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc
    13861 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
    13921 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
    13981 gatccctcag acccttttag tcagtgtgga aaatctctag cagtgccgcc cgaacaggga
    14041 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
    14101 gcgcacggca agaggcgaag ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
    14161 aaggctagaa ggaagaagat ggtgccagag cgtcagtatt aagcggggga aaattagatc
    14221 gcgaagggaa aaaattcggt taaggccagg
//
