LOCUS       sequence-259410-       15424 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express Firefly Luciferase-eGFP and Renilla Luciferase under 
            Ubiquitin-63E promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15424)
  AUTHORS   Buchman AB, Brogan DJ, Sun R, Yang T, Hsu PD, Akbari OS
  TITLE     Programmable RNA Targeting Using CasRx in Flies.
  JOURNAL   CRISPR J. 2020 Jun;3(3):164-176. doi: 10.1089/crispr.2020.0018.
  PUBMED    32584145
REFERENCE   2  (bases 1 to 15424)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1089/crispr.2020"; journalName: "CRISPR J"; date: "2020-06"; 
            volume: "3"; issue: "3"; pages: "164-176"
COMMENT     Sequence Label: OA-1052B (Ubiq-Fluc_Ubiq-Rluc)
FEATURES             Location/Qualifiers
     source          1..15424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(675..693)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(1145..1216,1618..1891,1966..2620,2682..2997,3201..3332,
                     3403..4017)
                     /codon_start=1
                     /gene="white"
                     /product="Drosophila white gene eye color pigment"
                     /label=mini-white
                     /note="This is a modified version of the white gene lacking
                     part of the first intron."
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     protein_bind    4792..4825
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             6750..8399
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(6816..6834)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             8400..8453
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(8454..8456,8457..8459,8460..9173)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTSKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8499..8520)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8760..8779)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9107..9128
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    9977..10058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10014..10033)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10068..10087
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(10101..11036)
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     protein_bind    12948..13017
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(13236..13254)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(13275..13291)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13275..13291)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13288..13310)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13299..13315
                     /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(13323..13329,13330..13347,13348..13353))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13368..13389
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13506..13523)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13677..14265)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13757..13776)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14436..15227,15228..15296))
                     /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     15059..15078
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15297..15401)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtacgtaccg
      661 ggcccctagt atgtatgtaa gttaataaaa cccatttttg cggaaagtag ataaaaaaaa
      721 catttttttt ttttactgca ctggatatca ttgaacttat ctgatcagtt ttaaatttac
      781 ttcgatccaa gggtatttga tgtaccaggt tctttcgatt acctctcact caaaatgaca
      841 ttccactcaa agtcagcgct gtttgcctcc ttctctgtcc acagaaatat cgccgtctct
      901 ttcgccgctg cgtccgctat ctctttcgcc accgtttgta gcgttacgta gcgtcaatgt
      961 ccgccttcag ttgcattttg tcagcggttt cgtgacgaag ctccaagcgg tttacgccat
     1021 caattaaaca caaagtgctg tgccaaaact cctctcgctt cttatttttg tttgtttttt
     1081 gagtgattgg ggtggtgatt ggttttgggt gggtaagcag gggaaagtgt gaaaaatccc
     1141 ggcaatgggc caagaggatc aggagctatt aattcgcgga ggcagcaaac acccatctgc
     1201 cgagcatctg aacaatgtga gtagtacatg tgcatacatc ttaagttcac ttgatctata
     1261 ggaactgcga ttgcaacatc aaattgtctg cggcgtgaga actgcgaccc acaaaaatcc
     1321 caaaccgcaa ttgcacaaac aaatagtgac acgaaacaga ttattctggt agctgttctc
     1381 gctatataag acaatttttg agatcatatc atgatcaaga catctaaagg cattcatttt
     1441 cgactatatt cttttttaca aaaaatataa caaccagata ttttaagctg atcctagatg
     1501 cacaaaaaat aaataaaagt ataaacctac ttcgtaggat acttcggggt actttttgtt
     1561 cggggttaga tgagcataac gcttgtagtt gatatttgag atcccctatc attgcagggt
     1621 gacagcggag cggcttcgca gagctgcatt aaccagggct tcgggcaggc caaaaactac
     1681 ggcacgctcc ggccacccag tccgccggag gactccggtt cagggagcgg ccaactagcc
     1741 gagaacctca cctatgcctg gcacaatatg gacatctttg gggcggtcaa tcagccgggc
     1801 tccggatggc ggcagctggt caaccggaca cgcggactat tctgcaacga gcgacacata
     1861 ccggcgccca ggaaacattt gctcaagaac ggtgagtttc tattcgcagt cggctgatct
     1921 gtgtgaaatc ttaataaagg gtccaattac caatttgaaa ctcagtttgc ggcgtggcct
     1981 atccgggcga acttttggcc gtgatgggca gttccggtgc cggaaagacg accctgctga
     2041 atgcccttgc ctttcgatcg ccgcagggca tccaagtatc gccatccggg atgcgactgc
     2101 tcaatggcca acctgtggac gccaaggaga tgcaggccag gtgcgcctat gtccagcagg
     2161 atgacctctt tatcggctcc ctaacggcca gggaacacct gattttccaa gccatggtgc
     2221 ggatgccacg acatctgacc tatcggcagc gagtggcccg cgtggatcag gtgatccagg
     2281 agctttcgct cagcaaatgt cagcacacga tcatcggtgt gcccggcagg gtgaaaggtc
     2341 tgtccggcgg agaaaggaag cgtctggcat tcgcctccga ggctctaacc gatccgccgc
     2401 ttctgatctg cgatgagccc acctccggac tggactcctt taccgcccac agcgtcgtcc
     2461 aggtgctgaa gaagctgtcg cagaagggca agaccgtcat cctgaccatt catcagccgt
     2521 cttccgagct gtttgagctc tttgacaaga tccttctgat ggccgagggc agggtagctt
     2581 tcttgggcac tcccagcgaa gccgtcgact tcttttccta gtgagttcga tgtgtttatt
     2641 aagggtatct agtattacat aacatctcaa ctcctatcca gcgtgggtgc ccagtgtcct
     2701 accaactaca atccggcgga cttttacgta caggtgttgg ccgttgtgcc cggacgggag
     2761 atcgagtccc gtgatcggat cgccaagata tgcgacaatt ttgccattag caaagtagcc
     2821 cgggatatgg agcagttgtt ggccaccaaa aatctggaga agccactgga gcagccggag
     2881 aatgggtaca cctacaaggc cacctggttc atgcagttcc gggcggtcct gtggcgatcc
     2941 tggctgtcgg tgctcaagga accactcctc gtaaaagtgc gacttattca gacaacggtg
     3001 agtggttcca gtggaaacaa atgatataac gcttacaatt cttggaaaca aattcgctag
     3061 attttagata gaattgcctg attccacacc cttcttagtt tttttcaatg agatgtatag
     3121 tttatagttt tgcagaagat aaataaattt catttaactc gcgaatatta atgagatgcg
     3181 agtaacattt taatttgcag atggttgcca tcttgattgg cctcatcttt ttgggccaac
     3241 aactcacgca agtgggtgtg atgaatatca acggagccat cttcctcttc ctgaccaaca
     3301 tgacctttca aaacgtcttt gccacgataa atgtaagtca tgtttagaat acatttgcat
     3361 ttcaataatt tactaacttt ctaatgaatc gattcgattt aggtgttcac ctcagagctg
     3421 ccagttttta tgagggaggc ccgaagtcga ctttatcgct gtgacacata ctttctgggc
     3481 aaaacgattg ccgaattgcc gctttttctc acagtgccac tggtcttcac ggcgattgcc
     3541 tatccgatga tcggactgcg ggccggagtg ctgcacttct tcaactgcct ggcgctggtc
     3601 actctggtgg ccaatgtgtc aacgtccttc ggatatctaa tatcctgcgc cagctcctcg
     3661 acctcgatgg cgctgtctgt gggtccgccg gttatcatac cattcctgct ctttggcggc
     3721 ttcttcttga actcgggctc ggtgccagta tacctcaaat ggttgtcgta cctctcatgg
     3781 ttccgttacg ccaacgaggg tctgctgatt aaccaatggg cggacgtgga gccgggcgaa
     3841 attagctgca catcgtcgaa caccacgtgc cccagttcgg gcaaggtcat cctggagacg
     3901 cttaacttct ccgccgccga tctgccgctg gactacgtgg gtctggccat tctcatcgtg
     3961 agcttccggg tgctcgcata tctggctcta agacttcggg cccgacgcaa ggagtagccg
     4021 acatatatcc gaaataactg cttgtttttt ttttttacca ttattaccat cgtgtttact
     4081 gtttattgcc ccctcaaaaa gctaatgtaa ttatatttgt gccaataaaa acaagatatg
     4141 acctatagaa tacaagtatt tccccttcga acatccccac aagtagactt tggatttgtc
     4201 ttctaaccaa aagacttaca cacctgcata ccttacatca aaaactcgtt tatcgctaca
     4261 taaaacaccg ggatatattt tttatataca tacttttcaa atcgcgcgcc ctcttcataa
     4321 ttcacctcca ccacaccacg tttcgtagtt gctctttcgc tgtctcccac ccgctctccg
     4381 caacacattc accttttgtt cgacgacctt ggagcgactg tcgttagttc cgcgcgattc
     4441 ggttcgctca aatggttccg agtggttcat ttcgtctcaa tagaaattag taataaatat
     4501 ttgtatgtac aatttatttg ctccaatata tttgtatata tttccctcac agctatattt
     4561 attctaattt aatattatga ctttttaagg taattttttg tgacctgttc ggagtgatta
     4621 gcgttacaat ttgaactgaa agtgacatcc agtgtttgtt ccttgtgtag atgcatctca
     4681 aaaaaatggt gggcataata gtgttgttta tatatatcaa aaataacaac tataataata
     4741 agaatacatt taatttagaa aatgcttgga tttcactgga actaggctag cataacttcg
     4801 tataatgtat gctatacgaa gttatgctag cggatccggg aattgggaat tgggcaatat
     4861 ttaaatggcg gctgcagcgc gcagatcgcc gatgggcgtg gcgccggagg gcaccttcac
     4921 cttgccgtac ggcttgctgt tcttcgcgtt caaaatctcc agctccattt tgctttcggt
     4981 gcgcttgcaa tcagtactgt ccaaaatcga aaatcgccga accgtagtgt gaccgtgcgg
     5041 ggctctgcga aaataaactt ttttaggtat atggccacac acggggaaag cacagtggat
     5101 tatatgtttt aatattataa tatgcaggtt ttcattactt atccagatgt aagcccactt
     5161 aaagcgattt aacaattatt tgccgaaaga gtataaacaa atttcactta aaaatggatt
     5221 aagaaaagct tgtgtaagat tatgcgcagc gttgccagat agctccattt aaaacacttc
     5281 aaaaacaata agttttgaaa atatatacat aaatagcagt cgttgccgca acgctcaaca
     5341 catcacactt ttaaaacacc ctttacctac acagaattac tttttaaatt tccagtcaag
     5401 ctgcgagttt caaaattata gccggtagag aagacagtgc tatttcaaaa gcaaactaac
     5461 aagggtctta aattccaaaa caccaatcct aacaagcctt ggacttttgt aagtttagat
     5521 caaaggtggc attgcattca atgtcatggt aagaagtagg tcgtctaggt agaaatcctc
     5581 attcagccgg tcaagtcagt acgagaaagg tctcaatttg aaattgtctt aaaaatattt
     5641 tattgttttg tactgtggtg agtttaaacg aaaaacacaa aaaaaaagtg atacacagaa
     5701 atcataaaaa attttaatac aaggtattcg tacgtatcaa aaacatttcg gcacaatttt
     5761 ttttctctgt actaaagtgt tacgaacact acggtatttt ttagtgattt tcaacggaca
     5821 ccgaaggtat ataaacagcg ttcgcgaacg gtcgccttca aaaccaattg acatttgcag
     5881 cagcaagtac aagtagaaag taaagcgcaa tcagcgaaaa atttatactt aattgttggt
     5941 gattaaagta caattaaaag aacattctcg aaagtcacaa gaaacgtaag tttttaactc
     6001 gctgttacca attagtaata agagcaacaa gacgttgagt aatttcaaga aaaactgcat
     6061 ttcaaggtct ttgttcggcc attttttttt tattcaacgc tctacgtaat tacaaaataa
     6121 gaaattggca gccacgcatc ttgttttccc aatgaattgg catcaaaacg caaacaaatc
     6181 tataaataaa acttgcgtgt tgattttcgc caagatttat tggcaaattg tgaaattcgc
     6241 agtgacgcat ttgaaaattc gagaaatcac gaacgcacat attgcatttg tgtgcatgtt
     6301 attagttagt tgtttagtta attgaagtat tttaccaacg aaatccactt atttttagct
     6361 gaaatagagt aggttgctta aacaaagcca cgtctgaaaa tttcttattg cttgtagttg
     6421 tgacgtcacc atatacacac aaaataatgt gtatgcatgc atttcagctg tgtatatata
     6481 catgcacaca ctcgcaacac gaaaacgatg acgaagcaac ggaacaaagg tttctcaact
     6541 accctttgtt ccctgtttct tcgctttcct ttgttccaat attcgtagag ggttaatagg
     6601 ggtttctcaa caaagttggc gtcgataaat aagtttccca tttttattcc ccagccagga
     6661 agttagtttc aatagttttg taatttcaac gaaactcatt tgatttcgta ctaattttcc
     6721 acatctctat tttgacccgc agacctagga tggaagacgc caaaaacata aagaaaggcc
     6781 cggcgccatt ctatccgctg gaagatggaa ccgctggaga gcaactgcat aaggctatga
     6841 agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc gaggtggaca
     6901 tcacttacgc tgagtacttc gaaatgtccg ttcggttggc agaagctatg aaacgatatg
     6961 ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc
     7021 cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac atttataatg
     7081 aacgtgaatt gctcaacagt atgggcattt cgcagcctac cgtggtgttc gtttccaaaa
     7141 aggggttgca aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa aaaattatta
     7201 tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc gtcacatctc
     7261 atctacctcc cggttttaat gaatacgatt ttgtgccaga gtccttcgat agggacaaga
     7321 caattgcact gatcatgaac tcctctggat ctactggtct gcctaaaggt gtcgctctgc
     7381 ctcatagaac tgcctgcgtg agattctcgc atgccagaga tcctattttt ggcaatcaaa
     7441 tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt ggaatgttta
     7501 ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga tttgaagaag
     7561 agctgtttct gaggagcctt caggattaca agattcaaag tgcgctgctg gtgccaaccc
     7621 tattctcctt cttcgccaaa agcactctga ttgacaaata cgatttatct aatttacacg
     7681 aaattgcttc tggtggcgct cccctctcta aggaagtcgg ggaagcggtt gccaagaggt
     7741 tccatctgcc aggtatcagg caaggatatg ggctcactga gactacatca gctattctga
     7801 ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca ttttttgaag
     7861 cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcaaaga ggcgaactgt
     7921 gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa tccggaagcg accaacgcct
     7981 tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac gaagacgaac
     8041 acttcttcat cgttgaccgc ctgaagtctc tgattaagta caaaggctat caggtggctc
     8101 ccgctgaatt ggaatccatc ttgctccaac accccaacat cttcgacgca ggtgtcgcag
     8161 gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa
     8221 agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa
     8281 agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg
     8341 acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag atcgccgtgg
     8401 agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc cctatggtga
     8461 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     8521 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     8581 tgaccctgaa gttcatctgc accaccagca agctgcccgt gccctggccc accctcgtga
     8641 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     8701 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     8761 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     8821 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     8881 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     8941 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     9001 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     9061 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     9121 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taagcgatcg
     9181 cctagaatga atcgttttta aaataacaaa tcaattgttt tataatattc gtacgattct
     9241 ttgattatgt aataaaatgt gatcattagg aagattacga aaaatataaa aaatatgagt
     9301 tctgtgtgta taacaaatgc tgtaaacgcc acaattgtgt ttgttgcaaa taaacccatg
     9361 attatttgat taaaattgtt gttttctttg ttcatagaca atagtgtgtt ttgcctaaac
     9421 gtgtactgca taaactccat gcgagtgtat agcgagctag tggctaacgc ttgccccacc
     9481 aaagtagatt cgtcaaaatc ctcaatttca tcaccctcct ccaagtttaa catttggccg
     9541 tcggaattaa cttctaaaga tgccacataa tctaataaat gaaatagaga ttcaaacgtg
     9601 gcgtcatcgt ccgtttcgac catttccgaa aagaactcgg gcataaactc tatgatttct
     9661 ctggacgtgg tgttgtcgaa actctcaaag tacgcagtca ggaacgtgcg cgacatgtcg
     9721 tcgggaaact cgcgcggaaa catgttgttg taaccgaacg ggtcccatag cgccaaaacc
     9781 aaatctgcca gcgtcaatag aatgagcacg atgccgacaa tggagctggc ttggatagcg
     9841 attcgagtta acggccctta agggtcgact gatcataatc agccatacca catttgtaga
     9901 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     9961 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
    10021 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
    10081 actcatcaat gtggcgcgcc ttattgttca tttttgagaa ctcgctcaac gaacgatttg
    10141 atatattttc ccatttcatc aggtgcatct tcttgcgaaa aatgaagacc ttttactttg
    10201 acaaattcag tattaggaaa cttcttggca ccttcaacaa tagcattgga aaagaatcct
    10261 gggtccgatt caataaacat ttttggtaaa tcatcacttg cacgtagata agcattataa
    10321 ttcctaacaa tttgtacaac gtcaggttta ccacctttta ctaacgggat ttcacgaggc
    10381 catgataatg ttggacgacg aacttcacct ttctctttga atggttcaag atatgctgca
    10441 aattcttctg gttctaactt tctcatgatt tttgatggca acatggtttc cacgaagaag
    10501 ttattctcca aaaccatttt ttctccttct tcagatttga tcaacgcaat atcttcttca
    10561 atatcaggcc attcatccca tgattcaatc acatctacta cactttcagc gtgaactatt
    10621 gctttgatct tatcttgatg ctcatagcta taatgaaatg ccaaacaagc accccaatca
    10681 tggccgacaa aaatgatctt ctttggtaaa ttaagaagtt caaaccatgc agtaagatat
    10741 ttgtaatgat caagtaacct ataagaacca ttaccagatt tgcctgattt gcccatacca
    10801 ataaggtctg gtataataca ccgcgctact ggctcaatat gtggcacaac atgtcgccat
    10861 aaataagaag aggccgcgtt accatgtaaa aaaataacag cattttctgc atgtttttct
    10921 gaatcataat aattaataaa tgaatcaaga acattcattt gtttacatct ggcccaccac
    10981 tgcggaccag ttatcatccg tttcctttgt tctggatcat aaactttcga agtcatgcgg
    11041 ccgctctgcg ggtcaaaata gagatgtgga aaattagtac gaaatcaaat gagtttcgtt
    11101 gaaattacaa aactattgaa actaacttcc tggctgggga ataaaaatgg gaaacttatt
    11161 tatcgacgcc aactttgttg agaaacccct attaaccctc tacgaatatt ggaacaaagg
    11221 aaagcgaaga aacagggaac aaagggtagt tgagaaacct ttgttccgtt gcttcgtcat
    11281 cgttttcgtg ttgcgagtgt gtgcatgtat atatacacag ctgaaatgca tgcatacaca
    11341 ttattttgtg tgtatatggt gacgtcacaa ctacaagcaa taagaaattt tcagacgtgg
    11401 ctttgtttaa gcaacctact ctatttcagc taaaaataag tggatttcgt tggtaaaata
    11461 cttcaattaa ctaaacaact aactaataac atgcacacaa atgctcgagt gcgttcgtga
    11521 tttctcgaat tttcaaatgc gtcactgcga atttcacaat ttgccaataa atcttggcga
    11581 aaatcaacac gcaagtttta tttatagatt tgtttgcgtt ttgatgccaa ttcattggga
    11641 aaacaagatg cgtggctgcc aatttcttat tttgtaatta cgtagagcgt tgaataaaaa
    11701 aaaaatggcc gaacaaagac cttgaaatgc agtttttctt gaaattactc aacgtcttgt
    11761 tgctcttatt actaattggt aacagcgagt taaaaactta cgtttcttgt gactttcgag
    11821 aatgttcttt taattgtact ttaatcacca acaattaagt ataaattttt cgctgattgc
    11881 gctttacttt ctacttgtac ttgctgctgc aaatgtcaat tggttttgaa ggcgaccgtt
    11941 cgcgaacgct gtttatatac cttcggtgtc cgttgaaaat cactaaaaaa taccgtagtg
    12001 ttcgtaacac tttagtacag agaaaaaaaa ttgtgccgaa atgtttttga tacgtacgaa
    12061 taccttgtat taaaattttt tatgatttct gtgtatcact ttttttttgt gtttttcgtt
    12121 taaactcacc acagtacaaa acaataaaat atttttaaga caatttcaaa ttgagacctt
    12181 tctcgtactg acttgaccgg ctgaatgagg atttctacct agacgaccta cttcttacca
    12241 tgacattgaa tgcaatgcca cctttgatct aaacttacaa aagtccaagg cttgttagga
    12301 ttggtgtttt ggaatttaag acccttgtta gtttgctttt gaaatagcac tgtcttctct
    12361 accggctata attttgaaac tcgcagcttg actggaaatt taaaaagtaa ttctgtgtag
    12421 gtaaagggtg ttttaaaagt gtgatgtgtt gagcgttgcg gcaacgactg ctatttatgt
    12481 atatattttc aaaacttatt gtttttgaag tgttttaaat ggagctatct ggcaacgctg
    12541 cgcataatct tacacaagct tttcttaatc catttttaag tgaaatttgt ttatactctt
    12601 tcggcaaata attgttaaat cgctttaagt gggcttacat ctggataagt aatgaaaacc
    12661 tgcatattat aatattaaaa catataatcc actgtgcttt ccccgtgtgt ggccatatac
    12721 ctaaaaaagt ttattttcgc agagccccgc acggtcacac tacggttcgg cgattttcga
    12781 ttttggacag tactgattgc aagcgcaccg aaagcaaaat ggagctggag attttgaacg
    12841 cgaagaacag caagccgtac ggcaaggtga aggtgccctc cggcgccacg cccatcggcg
    12901 atctgcgcgt ctagagatcc actagtgtcg acgatgtagg tcacggtctc gaagccgcgg
    12961 tgcgggtgcc agggcgtgcc cttgggctcc ccgggcgcgt actccacctc acccatctgg
    13021 tccatcatga tgaacgggtc gaggtggcgg tagttgatcc cggcgaacgc gcggcgcacc
    13081 gggaagccct cgccctcgaa accgctgggc gcggtggtca cggtgagcac gggacgtgcg
    13141 acggcgtcgg cgggtgcgga tacgcggggc agcgtcagcg ggttctcgac ggtcacggcg
    13201 ggcatgtcga cactagttct agccagcttt tgttcccttt agtgagggtt aatttcgagc
    13261 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
    13321 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
    13381 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
    13441 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
    13501 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
    13561 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
    13621 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
    13681 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
    13741 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
    13801 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
    13861 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
    13921 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
    13981 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
    14041 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
    14101 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
    14161 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
    14221 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
    14281 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
    14341 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
    14401 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
    14461 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
    14521 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
    14581 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
    14641 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
    14701 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
    14761 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
    14821 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
    14881 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
    14941 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
    15001 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
    15061 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
    15121 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
    15181 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
    15241 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
    15301 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
    15361 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
    15421 ccac
//
