LOCUS       sequence-344886-       11444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses humanized firefly luciferase and mCherry and Hygromycin 
            resistance linked by T2A, each controlled by a strong promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11444)
  TITLE     pLV[Expression]-mCherry:T2A:Hygro-EF1A>Luc2
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11444)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pLV[Expression]-mCherry:T2A:Hygro-EF1A>Luc2
FEATURES             Location/Qualifiers
     source          1..11444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /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    1803..1920
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    1938..1958
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        1959..3137
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2190..3128
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3085..3105
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    3137..3161
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      3162..3171
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3168..4820
                     /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"
     protein_bind    complement(4821..4845)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4863..5451
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4916..4936)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5334..5345)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     enhancer        5481..5860
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5861..6064
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6015..6035
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6091..6100
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6097..6804
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(6242..6260)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6510..6529
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             6814..6867
                     /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             6868..7893
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             6868..7893
                     /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"
     LTR             7965..8198
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8270..8391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8307..8326)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8361..8380
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8431..8566
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8586..8605)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8587..8605)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8615..8632)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8615..8631)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8624..8646)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8773..9228
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8860..8879)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9070..9091
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9254..9358
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9359..9427,9428..10219)
                     /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(9577..9596)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10390..10978
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10879..10898
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11132..11149
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11266..11287
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11302..11307,11308..11325,11326..11332)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11340..11356
                     /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)."
     primer_bind     11345..11367
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11364..11380
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11364..11380
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11399..11419
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11401..11419
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgcta
     1801 gcttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
     1861 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt
     1921 actagtgatt atcggatcaa ctttgtatag aaaagttggg ctccggtgcc cgtcagtggg
     1981 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg
     2041 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     2101 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     2161 ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg
     2221 gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg
     2281 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt
     2341 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg
     2401 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt
     2461 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg
     2521 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg
     2581 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac
     2641 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg tctcgcgccg ccgtgtatcg
     2701 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc
     2761 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg
     2821 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg
     2881 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta
     2941 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg
     3001 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt
     3061 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc
     3121 catttcaggt gtcgtgacaa gtttgtacaa aaaagcaggc tgccaccatg gaagatgcca
     3181 aaaacattaa gaagggccca gcgccattct acccactcga agacgggacc gccggcgagc
     3241 agctgcacaa agccatgaag cgctacgccc tggtgcccgg caccatcgcc tttaccgacg
     3301 cacatatcga ggtggacatt acctacgccg agtacttcga gatgagcgtt cggctggcag
     3361 aagctatgaa gcgctatggg ctgaatacaa accatcggat cgtggtgtgc agcgagaata
     3421 gcttgcagtt cttcatgccc gtgttgggtg ccctgttcat cggtgtggct gtggccccag
     3481 ctaacgacat ctacaacgag cgcgagctgc tgaacagcat gggcatcagc cagcccaccg
     3541 tcgtattcgt gagcaagaaa gggctgcaaa agatcctcaa cgtgcaaaag aagctaccga
     3601 tcatacaaaa gatcatcatc atggatagca agaccgacta ccagggcttc caaagcatgt
     3661 acaccttcgt gacttcccat ttgccacccg gcttcaacga gtacgacttc gtgcccgaga
     3721 gcttcgaccg ggacaaaacc atcgccctga tcatgaacag tagtggcagt accggattgc
     3781 ccaagggcgt agccctaccg caccgcaccg cttgtgtccg attcagtcat gcccgcgacc
     3841 ccatcttcgg caaccagatc atccccgaca ccgctatcct cagcgtggtg ccatttcacc
     3901 acggcttcgg catgttcacc acgctgggct acttgatctg cggctttcgg gtcgtgctca
     3961 tgtaccgctt cgaggaggag ctattcttgc gcagcttgca agactataag attcaatctg
     4021 ccctgctggt gcccacacta tttagcttct tcgctaagag cactctcatc gacaagtacg
     4081 acctaagcaa cttgcacgag atcgccagcg gcggggcgcc gctcagcaag gaggtaggtg
     4141 aggccgtggc caaacgcttc cacctaccag gcatccgcca gggctacggc ctgacagaaa
     4201 caaccagcgc cattctgatc acccccgaag gggacgacaa gcctggcgca gtaggcaagg
     4261 tggtgccctt cttcgaggct aaggtggtgg acttggacac cggtaagaca ctgggtgtga
     4321 accagcgcgg cgagctgtgc gtccgtggcc ccatgatcat gagcggctac gttaacaacc
     4381 ccgaggctac aaacgctctc atcgacaagg acggctggct gcacagcggc gacatcgcct
     4441 actgggacga ggacgagcac ttcttcatcg tggaccggct gaagagcctg atcaaataca
     4501 agggctacca ggtagcccca gccgaactgg agagcatcct gctgcaacac cccaacatct
     4561 tcgacgccgg ggtcgccggc ctgcccgacg acgatgccgg cgagctgccc gccgcagtcg
     4621 tcgtgctgga acacggtaaa accatgaccg agaaggagat cgtggactat gtggccagcc
     4681 aggttacaac cgccaagaag ctgcgcggtg gtgttgtgtt cgtggacgag gtgcctaaag
     4741 gactgaccgg caagttggac gcccgcaaga tccgcgagat tctcattaag gccaagaagg
     4801 gcggcaagat cgccgtgtaa acccagcttt cttgtacaaa gtggtgataa tcgaattccg
     4861 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     4921 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     4981 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     5041 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     5101 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     5161 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     5221 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     5281 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     5341 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     5401 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgggaat
     5461 tcccgcggtt cgaacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
     5521 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     5581 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     5641 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     5701 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     5761 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     5821 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     5881 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     5941 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     6001 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     6061 agctctctgg ctaactagag aacccactgc gccaccatgg tgagcaaggg cgaggaggat
     6121 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     6181 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     6241 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
     6301 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
     6361 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     6421 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
     6481 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     6541 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     6601 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
     6661 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
     6721 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
     6781 ggcggcatgg acgagctgta caagggctcc ggagagggca ggggaagtct tctaacatgc
     6841 ggggacgtgg aggaaaatcc cggccccatg aaaaagcctg aactcaccgc gacgtctgtc
     6901 gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc
     6961 gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat
     7021 agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg
     7081 ctcccgattc cggaagtgct tgacattggg gaatttagcg agagcctgac ctattgcatc
     7141 tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt
     7201 ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc
     7261 gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata
     7321 tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt
     7381 gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc
     7441 cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata
     7501 acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac
     7561 atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg
     7621 aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt
     7681 gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt
     7741 cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc
     7801 agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga
     7861 cgccccagca ctcgtccgag ggcaaaggaa tagggtacct ttaagaccaa tgacttacaa
     7921 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     7981 ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
     8041 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     8101 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     8161 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat
     8221 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat
     8281 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     8341 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     8401 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     8461 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     8521 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg gacgtaccca
     8581 attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg
     8641 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     8701 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     8761 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     8821 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     8881 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     8941 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     9001 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     9061 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     9121 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     9181 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     9241 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     9301 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     9361 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     9421 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     9481 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     9541 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     9601 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     9661 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     9721 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     9781 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     9841 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     9901 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     9961 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
    10021 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
    10081 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
    10141 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
    10201 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
    10261 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
    10321 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
    10381 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
    10441 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
    10501 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
    10561 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
    10621 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
    10681 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
    10741 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
    10801 tcccgaagag agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
    10861 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
    10921 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
    10981 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
    11041 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
    11101 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
    11161 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
    11221 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
    11281 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
    11341 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc
    11401 aattaaccct cactaaaggg aacaaaagct ggagctgcaa gctt
//
