pCLucIPZ

LOCUS       pCLucIPZ               12370 bp ds-DNA     circular SYN 10-JAN-2018
DEFINITION  Constitutive expression of Cypridina luciferase, puromycin 
            N-acetyl-transferase and shRNAmir of interest.
ACCESSION   .
VERSION     .
KEYWORDS    pCLucIPZ
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12370)
  TITLE     Dual secreted luciferase-based shRNA competition assay
  JOURNAL   Unpublished
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|biofeng.com|
COMMENT     VNTNAME|pCLucIPZ|
FEATURES             Location/Qualifiers
     source          1..12370
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2063..2180
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(2245..2619)
                     /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      2616..2625
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        complement(2629..2647)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2639..2686)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     enhancer        2791..3094
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3095..3298
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3249..3269
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3370..5031
                     /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"
     misc_feature    5048..5620
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5215..5232)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5442..5461
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5623..6225
                     /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     6122..6142
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     ncRNA           6232..6353
                     /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           6381..6395
                     /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           6423..6549
                     /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"
     primer_bind     complement(6643..6662)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6644..6662)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    6682..7270
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6735..6755)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7153..7164)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7477..7710
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7811..7828)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7817..8041
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8087..8515
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8174..8193)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8384..8405
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8524..8544)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8529..8858
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8709..8844
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8771..8790
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    9725..9846
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9895..9911)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9895..9911)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9908..9930)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9919..9935
                     /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(9943..9973)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9988..10009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10126..10143)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10297..10885)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10377..10396)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11056..11916)
                     /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     11679..11698
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11917..12021)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    12069..12203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
     2041 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     2101 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     2161 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag
     2221 taccgggccc gctctagtcc ggaatcagtc ctgctcctcg gccacgaagt gcacgcagtt
     2281 gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat
     2341 ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag
     2401 ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg
     2461 gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac
     2521 gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg
     2581 cgcggtgagc accggaacgg cactggtcaa cttggccatg gtggccctcc tatagtgagt
     2641 cgtattatac tatgccgata tactatgccg atgattaatt gtcaacacgt gctgcaggtc
     2701 cgaggttcta gacgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
     2761 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     2821 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     2881 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     2941 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     3001 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     3061 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     3121 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     3181 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     3241 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
     3301 tttagtgaac cgtcagatcc gctagcgcta ccgggcctgc aggttggtac cgagctcgga
     3361 tccgccacca tgaagacctt aattcttgcc gttgcattag tctactgcgc cactgttcat
     3421 tgccaggact gtccttacga acctgatcca ccaaacacag ttccaacttc ctgtgaagct
     3481 aaagaaggag aatgtattga tagcagctgt ggcacctgca cgagagacat actatcagat
     3541 ggactgtgtg aaaataaacc aggaaaaaca tgttgccgaa tgtgtcagta tgtaattgaa
     3601 tgcagagtag aggccgcagg atggtttaga acattctatg gaaagagatt ccagttccag
     3661 gaacctggta catacgtgtt gggtcaagga accaagggcg gcgactggaa ggtgtccatc
     3721 accctggaga acctggatgg aaccaagggg gctgtgctga ccaagacaag actggaagtg
     3781 gctggagaca tcattgacat cgctcaagct actgagaatc ccatcactgt aaacggtgga
     3841 gctgacccta tcatcgccaa cccgtacacc atcggcgagg tcaccatcgc tgttgttgag
     3901 atgccaggct tcaacatcac cgtcattgag ttcttcaaac tgatcgtgat cgacatcctc
     3961 ggaggaagat ctgtaagaat cgccccagac acagcaaaca aaggaatgat ctctggcctc
     4021 tgtggagatc ttaaaatgat ggaagataca gacttcactt cagatccaga acaactcgct
     4081 attcagccta agatcaacca ggagtttgac ggttgtccac tctatggaaa tcctgatgac
     4141 gttgcatact gcaaaggtct tctggagccg tacaaggaca gctgccgcaa ccccatcaac
     4201 ttctactact acaccatctc ctgcgccttc gcccgctgta tgggtggaga cgagcgagcc
     4261 tcacacgtgc tgcttgacta cagggagacg tgcgctgctc ccgaaactag aggaacctgc
     4321 gttttgtctg gacatacttt ctacgataca tttgacaaag caagatacca attccagggt
     4381 ccctgcaagg agattcttat ggccgccgac tgtttctgga acacttggga tgtgaaggtt
     4441 tcacacagga atgttgactc ttacactgaa gtagagaaag tacgaatcag gaaacaatcg
     4501 actgtagtag aactcattgt tgatggaaaa cagattctgg ttggaggaga agccgtgtcc
     4561 gtcccgtaca gctctcagaa cacttccatc tactggcaag atggtgacat actgactaca
     4621 gccatcctac ctgaagctct ggtggtcaag ttcaacttca agcaactgct cgtcgtacat
     4681 attagagatc cattcgatgg taagacttgc ggtatttgcg gtaactacaa ccaggatttc
     4741 agtgatgatt cttttgatgc tgaaggagcc tgtgatctga cccccaaccc accgggatgc
     4801 accgaagaac agaaacctga agctgaacga ctctgcaata gtctcttcgc cggtcaaagt
     4861 gatcttgatc agaaatgtaa cgtgtgccac aagcctgacc gtgtcgaacg atgcatgtac
     4921 gagtattgcc tgaggggaca acagggtttc tgtgaccacg catgggagtt caagaaagaa
     4981 tgctacataa agcatggaga caccctagaa gtaccagatg aatgcaaata ggcggccgca
     5041 aattccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     5101 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     5161 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     5221 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     5281 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     5341 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     5401 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     5461 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     5521 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     5581 accacgggga cgtggttttc ctttgaaaaa cacgataata ccatggccac cgagtacaag
     5641 cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc
     5701 gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg
     5761 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     5821 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     5881 gcggtgttcg ccgagatcgg ctcgcgcatg gccgagttga gcggttcccg gctggccgcg
     5941 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     6001 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     6061 cccggagtgg aggcggccga gcgcgctggg gtgcccgcct tcctggagac ctccgcgccc
     6121 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     6181 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtttg tttgaatgag
     6241 gcttcagtac tttacagaat cgttgcctgc acatcttgga aacacttgct gggattactt
     6301 cttcaggtta acccaacaga aggctcgaga aggtatattg ctgttgacag tgagcgatct
     6361 cgcttgggcg agagtaagta gtgaagccac agatgtactt actctcgccc aagcgagagt
     6421 gcctactgcc tcggaattca aggggctact ttaggagcaa ttatcttgtt tactaaaact
     6481 gaataccttg ctatctcttt gatacatttt tacaaagctg aattaaaatg gtataaatta
     6541 aatcactttt ttcaattgga agactaatgc ggccggccat tactccgtct cgtgtcttgt
     6601 tgcatatgtc tgctggtttg tttgatgttg tttgcgggcg ggccctatag tgagtcgtat
     6661 tacctaggac gcgtctggaa caatcaacct ctggattaca aaatttgtga aagattgact
     6721 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     6781 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     6841 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     6901 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     6961 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     7021 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     7081 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     7141 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     7201 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     7261 gcctccccgc ctggaattaa ttctgcagtc gagacctaga aaaacatgga gcaatcacaa
     7321 gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg
     7381 aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg
     7441 tagatcttag ccacttttta aaagaaaaga ggggactgga agggctaatt cactcccaac
     7501 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     7561 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     7621 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     7681 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     7741 atttataact tgcaaagaaa tgaatatcag agagtgagag gccttgacat tgtttaaacc
     7801 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     7861 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     7921 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     7981 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     8041 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     8101 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     8161 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     8221 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     8281 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     8341 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     8401 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     8461 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     8521 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     8581 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     8641 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     8701 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     8761 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     8821 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     8881 ccattttcgg atctgatcag cacgtgatga aaaagcctga actcaccgcg acgtcgctgc
     8941 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     9001 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     9061 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     9121 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     9181 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     9241 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     9301 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc
     9361 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     9421 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     9481 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt
     9541 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagcacgt
     9601 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     9661 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     9721 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     9781 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     9841 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     9901 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     9961 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
    10021 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
    10081 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
    10141 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
    10201 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
    10261 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
    10321 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
    10381 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
    10441 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
    10501 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
    10561 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
    10621 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
    10681 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
    10741 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
    10801 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
    10861 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
    10921 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
    10981 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
    11041 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
    11101 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
    11161 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
    11221 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
    11281 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
    11341 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
    11401 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
    11461 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
    11521 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
    11581 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
    11641 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
    11701 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
    11761 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
    11821 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
    11881 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
    11941 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    12001 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
    12061 gggagatcaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
    12121 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
    12181 atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac
    12241 ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct
    12301 gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa
    12361 acttagtagt
//