pLenti HRE-Luc pGK Hygro

LOCUS       pLenti HRE-Luc pGK Hygro       10634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  reporter plasmid HIF-responsive promoter (3xHRE) wt- fused to 
            firefly luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10634)
  AUTHORS   Briggs KJ, Koivunen P, Cao S, Backus KM, Olenchock BA, Patel H, 
            Zhang Q, Signoretti S, Gerfen GJ, Richardson AL, Witkiewicz AK, 
            Cravatt BF, Clardy J, Kaelin WG Jr
  TITLE     Paracrine Induction of HIF by Glutamate in Breast Cancer: EglN1 
            Senses Cysteine.
  JOURNAL   Cell. 2016 Jun 30;166(1):126-39. doi: 10.1016/j.cell.2016.05.042.
  PUBMED    27368101
REFERENCE   2  (bases 1 to 10634)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2016.05"; journalName: "Cell"; date: "2016-06-30- 
            30"; volume: "166"; issue: "1"; pages: "126-39"
COMMENT     Sequence Label: pLenti HRE-Luc pGK Hygro
FEATURES             Location/Qualifiers
     source          1..10634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        87..390
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        392..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     545..565
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             608..788
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    835..960
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1457..1690
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1875..1919
                     /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    2217..2334
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2678..2740
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     regulatory      2798..2807
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2804..4456
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     
     primer_bind     complement(2870..2888)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     promoter        4551..5050
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4586..4609)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4983..5002
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             5068..6093
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             5068..6093
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     protein_bind    complement(6124..6157)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6213..6801
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6266..6286)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6684..6695)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6804..6820)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6805..6821)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7010..7243
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7315..7436
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7352..7371)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7406..7425
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7476..7611
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7631..7650)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7632..7650)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(7673..8031)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7713..7734)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7925..7944
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        8058..8162
                     /gene="bla"
                     /label=AmpR promoter
     promoter        8164..8521
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        8164..8355
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(8187..8207)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      8372..8507
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             8556..9350
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             8556..9350
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8610..8629)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9220..9239
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(9538..9557)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    9582..9629
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      9958..10546
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10447..10466
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta
       61 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      121 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      181 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      241 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      301 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      361 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      421 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      481 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      541 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc
      601 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
      661 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
      721 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
      781 ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct
      841 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg
      901 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc
      961 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga
     1021 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
     1081 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
     1141 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
     1201 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
     1261 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac
     1321 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
     1381 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
     1441 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
     1501 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
     1561 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
     1621 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
     1681 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
     1741 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
     1801 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     1861 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     1921 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     1981 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     2041 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     2101 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     2161 acggatcggc actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt
     2221 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     2281 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
     2341 tattacaggg acagcagaga tccagtttgg ttagtaccgg gcccgctcta gccctagatt
     2401 tgggctcgaa gccctacgtg ctgtctcaca cagcctgtct gacctctcga cctaccggcc
     2461 gttcgaagcc ctacgtgctg tctcacacag ccttctgatc tcgacctacc ggccgttcga
     2521 agccctacgt gctgtctcac acagcctgtc tgacctctcg acctaccggc cgttcgagtc
     2581 gacagctgta ctgcagagat ccggccccgc ccagcgtctt gtcattggcg aattcgaaca
     2641 cgcagatgca gtcggggcgg cgcggtccga ggtccacttc gcatattaag gtgacgcgtg
     2701 tggcctcgaa caccgagcga ccctgcagcg acccgcttaa cagcgtcaac agcgtgccgc
     2761 agatctccaa gcttggcatt ccggtactgt tggtaaagcc accatggaag acgccaaaaa
     2821 cataaagaaa ggcccggcgc cattctatcc gctggaagat ggaaccgctg gagagcaact
     2881 gcataaggct atgaagagat acgccctggt tcctggaaca attgctttta cagatgcaca
     2941 tatcgaggtg gacatcactt acgctgagta cttcgaaatg tccgttcggt tggcagaagc
     3001 tatgaaacga tatgggctga atacaaatca cagaatcgtc gtatgcagtg aaaactctct
     3061 tcaattcttt atgccggtgt tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa
     3121 cgacatttat aatgaacgtg aattgctcaa cagtatgggc atttcgcagc ctaccgtggt
     3181 gttcgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaagc tcccaatcat
     3241 ccaaaaaatt attatcatgg attctaaaac ggattaccag ggatttcagt cgatgtacac
     3301 gttcgtcaca tctcatctac ctcccggttt taatgaatac gattttgtgc cagagtcctt
     3361 cgatagggac aagacaattg cactgatcat gaactcctct ggatctactg gtctgcctaa
     3421 aggtgtcgct ctgcctcata gaactgcctg cgtgagattc tcgcatgcca gagatcctat
     3481 ttttggcaat caaatcattc cggatactgc gattttaagt gttgttccat tccatcacgg
     3541 ttttggaatg tttactacac tcggatattt gatatgtgga tttcgagtcg tcttaatgta
     3601 tagatttgaa gaagagctgt ttctgaggag ccttcaggat tacaagattc aaagtgcgct
     3661 gctggtgcca accctattct ccttcttcgc caaaagcact ctgattgaca aatacgattt
     3721 atctaattta cacgaaattg cttctggtgg cgctcccctc tctaaggaag tcggggaagc
     3781 ggttgccaag aggttccatc tgccaggtat caggcaagga tatgggctca ctgagactac
     3841 atcagctatt ctgattacac ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt
     3901 tccatttttt gaagcgaagg ttgtggatct ggataccggg aaaacgctgg gcgttaatca
     3961 aagaggcgaa ctgtgtgtga gaggtcctat gattatgtcc ggttatgtaa acaatccgga
     4021 agcgaccaac gccttgattg acaaggatgg atggctacat tctggagaca tagcttactg
     4081 ggacgaagac gaacacttct tcatcgttga ccgcctgaag tctctgatta agtacaaagg
     4141 ctatcaggtg gctcccgctg aattggaatc catcttgctc caacacccca acatcttcga
     4201 cgcaggtgtc gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt
     4261 tttggagcac ggaaagacga tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt
     4321 aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct
     4381 taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca agaagggcgg
     4441 aaagatcgcc gtgtaattct agactgccgg atcgaattgt cgacgaattc ggatccgcgg
     4501 ccgcgtttaa acttaattaa ggccggccct cgaggttaac gaattctacc gggtagggga
     4561 ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg
     4621 ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc
     4681 tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc
     4741 ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag
     4801 tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg
     4861 ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg
     4921 gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc
     4981 ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc
     5041 gggcctttcg acctgcatcc cgccaccatg aaaaagcctg aactcaccgc gacgtctgtc
     5101 gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc
     5161 gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat
     5221 agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg
     5281 ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc
     5341 tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt
     5401 ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc
     5461 gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata
     5521 tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt
     5581 gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc
     5641 cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata
     5701 acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac
     5761 atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg
     5821 aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt
     5881 gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt
     5941 cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc
     6001 agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga
     6061 cgccccagca ctcgtccgag ggcaaaggaa tagtgtacaa gtaggaattc gtcgagggac
     6121 ctaataactt cgtatagcat acattatacg aagttataca tgtttaaggg ttccggttcc
     6181 actaggtaca attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     6241 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     6301 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     6361 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     6421 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     6481 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     6541 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     6601 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     6661 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     6721 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     6781 ccctttgggc cgcctccccg catcgatacc gtcgacctcg atcgagacct agaaaaacat
     6841 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
     6901 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
     6961 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     7021 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     7081 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     7141 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     7201 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     7261 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     7321 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     7381 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     7441 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     7501 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     7561 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
     7621 acccaattcg ccctatagtg agtcgtatta gcatgtgagc aaaaggccag cataaatcaa
     7681 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     7741 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     7801 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     7861 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     7921 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     7981 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     8041 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     8101 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     8161 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     8221 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     8281 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     8341 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     8401 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     8461 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     8521 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     8581 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     8641 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     8701 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     8761 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     8821 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     8881 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     8941 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     9001 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     9061 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     9121 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     9181 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     9241 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     9301 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     9361 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     9421 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     9481 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     9541 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     9601 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     9661 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     9721 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     9781 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     9841 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     9901 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     9961 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
    10021 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
    10081 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
    10141 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
    10201 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
    10261 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
    10321 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
    10381 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
    10441 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
    10501 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
    10561 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
    10621 atcccctgat tctg
//