LOCUS       sequence-464745-       13526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ef1a downstream of a ubiquitous chromatin opening element drives 
            expression of Luciferase2 and IRES GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13526)
  AUTHORS   Khan KA, Qureshi AA, Xu P, Kuo HY, Schumacher TN, Michael IP, Kerbel
            RS
  TITLE     Immunological Tolerance to Luciferase and Fluorescent Proteins using
            Tol Mice Enables Development of Improved Tumor Models for 
            Investigating Immunity and Metastasis.
  JOURNAL   Cancer Res. 2025 Apr 14. doi: 10.1158/0008-5472.CAN-24-2418.
  PUBMED    40228139
REFERENCE   2  (bases 1 to 13526)
  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: "Cancer Res.
            2025 Apr 14. doi: 10.1158/0008-5472.CAN-24-2418."
COMMENT     Sequence Label: pLUEF1 Luc2-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..13526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    39..590
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(183..200)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     410..429
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(593..595,596..598,599..1309)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(593..595,596..598,599..1309)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(638..659)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(899..918)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1246..1267
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1388..1404)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1389..1405)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     misc_feature    1418..2006
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1471..1491)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1889..1900)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(2150..2183)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             2203..2383
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(2408..2426)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2409..2426)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2716..2735)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(2949..2968)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3068..3090
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3128..3146)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3214..3318
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3319..3387,3388..4179)
                     /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     complement(3537..3556)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4350..4938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4839..4858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5092..5109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5179..5199)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5184..5513
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5364..5499
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          6647..6712
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             6842..6862
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    7287..7421
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7324..7343)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7378..7397
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             7589..8222
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    8269..8394
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8889..9122
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9307..9351
                     /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    9636..9753
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        10485..11669
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          10716..11660
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     11617..11637
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    11717..11834
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             11867..13519
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
ORIGIN
        1 aaactacggg ctgcaggaat tccgcccccc cccccctaac gttactggcc gaagccgctt
       61 ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg
      121 caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc
      181 ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga
      241 agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc
      301 tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc
      361 acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc
      421 aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga
      481 tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa acgtctaggc
      541 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata ccatggtgag
      601 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
      661 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
      721 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
      781 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
      841 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
      901 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
      961 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1021 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     1081 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     1141 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1201 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1261 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt ccggactcag
     1321 atctcgacta gctagtagct agctagctag tcgagctcaa gcttcgaatt cgatatcaag
     1381 cttatcgcga taccgtcgac ctcgagggaa ttccgataat caacctctgg attacaaaat
     1441 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     1501 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     1561 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     1621 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     1681 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     1741 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     1801 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     1861 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     1921 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     1981 gatctccctt tgggccgcct ccccgcatcg ggaattcgag ctcggtacct ttaagaccaa
     2041 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     2101 ggctaattca ctcccaacga agacaagatg ggatcaattc accatgggaa taacttcgta
     2161 tagcatacat tatacgaagt tatgctgctt tttgcttgta ctgggtctct ctggttagac
     2221 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     2281 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     2341 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagcatcta gaattaattc
     2401 cgtgtattct atagtgtcac ctaaatcgta tgtgtatgat acataaggtt atgtattaat
     2461 tgtagccgcg ttctaacgac aatatgtaca agcctaattg tgtagcatct ggcttactga
     2521 agcagaccct atcatctctc tcgtaaactg ccgtcagagt cggtttggtt ggacgaacct
     2581 tctgagtttc tggtaacgcc gtcccgcacc cggaaatggt cagcgaacca atcagcaggg
     2641 tcatcgctag ccagatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag
     2701 gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact
     2761 tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac
     2821 tgttgggcgc catctccttg catgcaccat tccttgcggc ggcggtgctc aacggcctca
     2881 acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt cgaatggtgc
     2941 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     3001 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     3061 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     3121 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     3181 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3241 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3301 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3361 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3421 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3481 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3541 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3601 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3661 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3721 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3781 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3841 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3901 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3961 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4021 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4081 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4141 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4201 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4261 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4321 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4381 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4441 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4501 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4561 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4621 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4681 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4741 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4801 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4861 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4921 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4981 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5041 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     5101 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     5161 attcattaat gcagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     5221 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     5281 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5341 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     5401 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     5461 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg
     5521 gacacaagac aggcttgcga gatatgtttg agaataccac tttatcccgc gtcagggaga
     5581 ggcagtgcgt aaaaagacgc ggactcatgt gaaatactgg tttttagtgc gccagatctc
     5641 tataatctcg cgcaacctat tttcccctcg aacacttttt aagccgtaga taaacaggct
     5701 gggacacttc acatgagcga aaaatacatc gtcacctggg acatgttgca gatccatgca
     5761 cgtaaactcg caagccgact gatgccttct gaacaatgga aaggcattat tgccgtaagc
     5821 cgtggcggtc tgtaccgggt gcgttactgg cgcgtgaact gggtattcgt catgtcgata
     5881 ccgtttgtat ttccagctac gatcacgaca accagcgcga gcttaaagtg ctgaaacgcg
     5941 cagaaggcga tggcgaaggc ttcatcgtta ttgatgacct ggtggatacc ggtggtactg
     6001 cggttgcgat tcgtgaaatg tatccaaaag cgcactttgt caccatcttc gcaaaaccgg
     6061 ctggtcgtcc gctggttgat gactatgttg ttgatatccc gcaagatacc tggattgaac
     6121 agccgtggga tatgggcgtc gtattcgtcc cgccaatctc cggtcgctaa tcttttcaac
     6181 gcctggcact gccgggcgtt gttcttttta acttcaggcg ggttacaata gtttccagta
     6241 agtattctgg aggctgcatc catgacacag gcaaacctga gcgaaaccct gttcaaaccc
     6301 cgctttaaac atcctgaaac ctcgacgcta gtccgccgct ttaatcacgg cgcacaaccg
     6361 cctgtgcagt cggcccttga tggtaaaacc atccctcact ggtatcgcat gattaaccgt
     6421 ctgatgtgga tctggcgcgg cattgaccca cgcgaaatcc tcgacgtcca ggcacgtatt
     6481 gtgatgagcg atgccgaacg taccgacgat gatttatacg atacggtgat tggctaccgt
     6541 ggcggcaact ggatttatga gtgggccccg gatctttgtg aaggaacctt acttctgtgg
     6601 tgtgacataa ttggacaaac tacctacaga gatttaaagc tctaaggtaa atataaaatt
     6661 tttaagtgta taatgtgtta aactactgat tctaattgtt tgtgtatttt agattccaac
     6721 ctatggaact gatgaatggg agcagtggtg gaatgccttt aatgaggaaa acctgttttg
     6781 ctcagaagaa atgccatcta gtgatgatga ggctactgct gactctcaac attctactcc
     6841 tccaaaaaag aagagaaagg tagaagaccc caaggacttt ccttcagaat tgctaagttt
     6901 tttgagtcat gctgtgttta gtaatagaac tcttgcttgc tttgctattt acaccacaaa
     6961 ggaaaaagct gcactgctat acaagaaaat tatggaaaaa tattctgtaa cctttataag
     7021 taggcataac agttataatc ataacatact gttttttctt actccacaca ggcatagagt
     7081 gtctgctatt aataactatg ctcaaaaatt gtgtaccttt agctttttaa tttgtaaagg
     7141 ggttaataag gaatatttga tgtatagtgc cttgactaga gatcataatc agccatacca
     7201 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     7261 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     7321 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     7381 gtttgtccaa actcatcaat gtatcttatc atgtctggat caactggata actcaagcta
     7441 accaaaatca tcccaaactt cccaccccat accctattac cactgccaat tacctgtggt
     7501 ttcatttact ctaaacctgt gattcctctg aattattttc attttaaaga aattgtattt
     7561 gttaaatatg tactacaaac ttagtagttg gaagggctaa ttcactccca aagaagacaa
     7621 gatatccttg atctgtggat ctaccacaca caaggctact tccctgatta gcagaactac
     7681 acaccagggc caggggtcag atatccactg acctttggat ggtgctacaa gctagtacca
     7741 gttgagccag ataaggtaga agaggccaat aaaggagaga acaccagctt gttacaccct
     7801 gtgagcctgc atgggatgga tgacccggag agagaagtgt tagagtggag gtttgacagc
     7861 cgcctagcat ttcatcacgt ggcccgagag ctgcatccgg agtacttcaa gaactgctga
     7921 tatcgagctt gctacaaggg actttccgct ggggactttc cagggaggcg tggcctgggc
     7981 gggactgggg agtggcgagc cctcagatcc tgcatataag cagctgcttt ttgcctgtac
     8041 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     8101 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     8161 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     8221 cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc
     8281 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg
     8341 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt
     8401 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa
     8461 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
     8521 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
     8581 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
     8641 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
     8701 caaaacaaaa gtaagaccac cgcacagcaa gcgcggccgc tgatcttcag acctggagga
     8761 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     8821 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     8881 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     8941 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     9001 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     9061 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     9121 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     9181 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     9241 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     9301 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     9361 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     9421 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     9481 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     9541 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct
     9601 cgacggtatc gccaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     9661 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     9721 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     9781 ccagtttgga tcgataagct tgatatcgaa ttcgggaggt ggtccctgca gttacgccaa
     9841 tgataacccc cgccagaaaa atcttagtag ccttcccttt ttgttttccg tgccccaact
     9901 cggcggattg actcggcccc ttccggaaac acccgaatca acttctagtc aaattattgt
     9961 tcacgccgca atgacccacc cctggcccgc gtctgtggaa ctgacccctg gtgtacagga
    10021 gagttcgctg ctgaaagtgg tcccaaaggg gtactagttt ttaagctccc aactccccct
    10081 cccccagcgt ctggaggatt ccacaccctc gcaccgcagg ggcgaggaag tgggcggagt
    10141 ccggttttgg cgccagccgc tgaggctgcc aagcagaaaa gccaccgctg aggagactcc
    10201 ggtcactgtc ctcgccccgc ctcccccttc cctccccttg gggaccaccg ggcgccacgc
    10261 cgcgaacggt aagtgccgcg gtcgtcggcg cctccgccct ccccctaggg ccccaattcc
    10321 cagcgggcgc ggcgcgcggc ccctcccccc gccgggcgcg cgcccgctgc cccgcccttc
    10381 gtggccgccc ggcgtgggcg gtgccacccc tccccccggc ggccccgcgc gcagctcccg
    10441 gctccctccc ccttcggatg tggcttgagc tgtaggcgcg gaggggctcc ggtgcccgtc
    10501 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
    10561 gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
    10621 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
    10681 ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg
    10741 ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc acctggctgc
    10801 agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt
    10861 gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc
    10921 gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag
    10981 ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa
    11041 atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg
    11101 cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa
    11161 tcggacgggg gtagtctcaa gctggccggc cccggcctgc tctggtgcct ggcctcgcgc
    11221 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
    11281 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
    11341 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
    11401 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
    11461 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
    11521 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
    11581 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
    11641 gtttttttct tccatttcag gtgtcgtgag gaatttcgac atttaaattt aattaatctc
    11701 gacggtatcg gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa
    11761 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa
    11821 aaattcaaaa ttttatcgat cacgagacta gcctcgaggt ttaaacatgg aagatgccaa
    11881 aaacattaag aagggcccag cgccattcta cccactcgaa gacgggaccg ccggcgagca
    11941 gctgcacaaa gccatgaagc gctacgccct ggtgcccggc accatcgcct ttaccgacgc
    12001 acatatcgag gtggacatta cctacgccga gtacttcgag atgagcgttc ggctggcaga
    12061 agctatgaag cgctatgggc tgaatacaaa ccatcggatc gtggtgtgca gcgagaatag
    12121 cttgcagttc ttcatgcccg tgttgggtgc cctgttcatc ggtgtggctg tggccccagc
    12181 taacgacatc tacaacgagc gcgagctgct gaacagcatg ggcatcagcc agcccaccgt
    12241 cgtattcgtg agcaagaaag ggctgcaaaa gatcctcaac gtgcaaaaga agctaccgat
    12301 catacaaaag atcatcatca tggatagcaa gaccgactac cagggcttcc aaagcatgta
    12361 caccttcgtg acttcccatt tgccacccgg cttcaacgag tacgacttcg tgcccgagag
    12421 cttcgaccgg gacaaaacca tcgccctgat catgaacagt agtggcagta ccggattgcc
    12481 caagggcgta gccctaccgc accgcaccgc ttgtgtccga ttcagtcatg cccgcgaccc
    12541 catcttcggc aaccagatca tccccgacac cgctatcctc agcgtggtgc catttcacca
    12601 cggcttcggc atgttcacca cgctgggcta cttgatctgc ggctttcggg tcgtgctcat
    12661 gtaccgcttc gaggaggagc tattcttgcg cagcttgcaa gactataaga ttcaatctgc
    12721 cctgctggtg cccacactat ttagcttctt cgctaagagc actctcatcg acaagtacga
    12781 cctaagcaac ttgcacgaga tcgccagcgg cggggcgccg ctcagcaagg aggtaggtga
    12841 ggccgtggcc aaacgcttcc acctaccagg catccgccag ggctacggcc tgacagaaac
    12901 aaccagcgcc attctgatca cccccgaagg ggacgacaag cctggcgcag taggcaaggt
    12961 ggtgcccttc ttcgaggcta aggtggtgga cttggacacc ggtaagacac tgggtgtgaa
    13021 ccagcgcggc gagctgtgcg tccgtggccc catgatcatg agcggctacg ttaacaaccc
    13081 cgaggctaca aacgctctca tcgacaagga cggctggctg cacagcggcg acatcgccta
    13141 ctgggacgag gacgagcact tcttcatcgt ggaccggctg aagagcctga tcaaatacaa
    13201 gggctaccag gtagccccag ccgaactgga gagcatcctg ctgcaacacc ccaacatctt
    13261 cgacgccggg gtcgccggcc tgcccgacga cgatgccggc gagctgcccg ccgcagtcgt
    13321 cgtgctggaa cacggtaaaa ccatgaccga gaaggagatc gtggactatg tggccagcca
    13381 ggttacaacc gccaagaagc tgcgcggtgg tgttgtgttc gtggacgagg tgcctaaagg
    13441 actgaccggc aagttggacg cccgcaagat ccgcgagatt ctcattaagg ccaagaaggg
    13501 cggcaagatc gccgtgtaat aagttt
//
