LOCUS       sequence-380729-       10178 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation, Inducible lentiviral expression, TRE-gateway; 
            PGK-rtTA-2A-blasti.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10178)
  TITLE     Superti-Furga RESOLUTE modified pLIX plasmids
REFERENCE   2  (bases 1 to 10178)
  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"
COMMENT     Sequence Label: pCW57.1_V5_TurboID_FLAG_Ct
FEATURES             Location/Qualifiers
     source          1..10178
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    41..158
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        212..526
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    220..238
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    256..274
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    291..309
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    327..345
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    363..381
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    398..416
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    434..452
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     499..523
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    547..671
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(696..701,702..719,720..726)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             780..1439
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(846..865)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1781..2086
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1965..1984
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2127..2251)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2262..2285
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2316..3272
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     promoter        3321..3831
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3750..3768
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3841..4236
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             4237..4290
                     /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(4291..4908,4909..4914,4915..4950,4951..4953,4954..4989,
                     4990..4992,4993..5028,5029..5037)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(5077..5097)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5495..5506)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5711..5944
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6016..6137
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6053..6072)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6107..6126
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6177..6312
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6312..6383
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6384..6452,6453..7244)
                     /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"
     rep_origin      7415..8003
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7904..7923
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8157..8174
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8291..8312
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8327..8332,8333..8350,8351..8357)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8365..8381
                     /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     8370..8392
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8389..8405
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8389..8405
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8424..8444
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8426..8444
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8472..8698
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8699..8879
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8926..9051
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9544..9777
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9962..10006
                     /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"
ORIGIN
        1 aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
       61 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
      121 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
      181 gagatccact ttggccgcga atcgatatgt cgagtttact ccctatcagt gatagagaac
      241 gtatgtcgag tttactccct atcagtgata gagaacgatg tcgagtttac tccctatcag
      301 tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt
      361 actccctatc agtgatagag aacgtatgtc gagtttatcc ctatcagtga tagagaacgt
      421 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg
      481 ggaggcctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gaattggcta
      541 gcatcaacaa gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat
      601 atattaaatt agattttgca taaaaaacag actacataat actgtaaaac acaacatatc
      661 cagtcactat ggcggccgca ttaggcaccc caggctttac actttatgct tccggctcgt
      721 ataatgtgtg gattttgagt taggatccgt cgagattttc aggagctaag gaagctaaaa
      781 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
      841 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata
      901 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
      961 acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg aaagacggtg
     1021 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa
     1081 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt
     1141 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga
     1201 atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg
     1261 ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg
     1321 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg
     1381 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaaa
     1441 gatctggatc cggcttacta aaagccagat aacagtatgc gtatttgcgc gctgattttt
     1501 gcggtataag aatatatact gatatgtata cccgaagtat gtcaaaaaga ggtatgctat
     1561 gaagcagcgt attacagtga cagttgacag cgacagctat cagttgctca aggcatatat
     1621 gatgtcaata tctccggtct ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt
     1681 gccgaacgct ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg gtttattgaa
     1741 atgaacggct cttttgctga cgagaacagg ggctggtgaa atgcagttta aggtttacac
     1801 ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac
     1861 gcccgggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc
     1921 ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga
     1981 tatggccagt gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga
     2041 aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt caggctccct
     2101 tatacacagc cagtctgcag gtcgaccata gtgactggat atgttgtgtt ttacagtatt
     2161 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg
     2221 tttctcgttc agctttcttg tacaaagtgg ttgattctag agactacaaa gacgatgacg
     2281 ataaaccagc gggcggcggc cccggctctg gagggaaaga caatactgtg cctctgaagc
     2341 tgatcgctct cctggctaat ggcgagttcc atagtggcga acagctggga gaaaccctgg
     2401 gcatgtccag ggccgctatc aacaagcaca ttcagactct gcgcgactgg ggcgtggacg
     2461 tgttcaccgt gcccggaaag ggctactctc tgcccgagcc tatcccgctg ctgaacgcta
     2521 aacagattct gggacagctg gacggcggga gcgtggcagt cctgcctgtg gtcgactcca
     2581 ccaatcagta cctgctggat cgaatcggcg agctgaagag tggggatgct tgcattgcag
     2641 aatatcagca ggcagggaga ggaagcagag ggaggaaatg gttctctcct tttggagcta
     2701 acctgtacct gagtatgttt tggcgcctga agcggggacc agcagcaatc ggcctgggcc
     2761 cggtcatcgg aattgtcatg gcagaagcgc tgcgaaagct gggagcagac aaggtgcgag
     2821 tcaaatggcc caatgacctg tatctgcagg atagaaagct ggcaggcatc ctggtggagc
     2881 tggccggaat aacaggcgat gctgcacaga tcgtcattgg cgccgggatt aacgtggcta
     2941 tgaggcgcgt ggaggaaagc gtggtcaatc agggctggat cacactgcag gaagcaggga
     3001 ttaacctgga caggaatact ctggccgcta cgctgatccg agagctgcgg gcagccctgg
     3061 aactgttcga gcaggaaggc ctggctccat atctgccacg gtgggagaag ctggataact
     3121 tcatcaatag acccgtgaag ctgatcattg gggacaaaga gattttcggg attagccggg
     3181 ggattgataa acagggagcc ctgctgctgg aacaggacgg agttatcaaa ccctggatgg
     3241 gcggagaaat cagtctgcgg tctgccgaaa agggacccat ccccaacccc ctgctgggcc
     3301 tggacagcac ctaaggatcc ggggttgggg ttgcgccttt tccaaggcag ccctgggttt
     3361 gcgcagggac gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg
     3421 tctcgcacat tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg
     3481 ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg
     3541 gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc agacggacag
     3601 cgccagggag caatggcagc gcgccgaccg cgatgggctg tggccaatag cggctgctca
     3661 gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg
     3721 gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc
     3781 acgtcggcag tcggctccct cgttgaccga atcaccgacc tctctcccca gcaattcacc
     3841 atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc
     3901 aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc
     3961 cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg
     4021 gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga
     4081 aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg
     4141 catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt
     4201 cgtgaattgc tgccctctgg ttatgtgtgg gagggcgaag gtagaggttc tctcctcact
     4261 tgtggtgatg ttgaagaaaa ccctggtcca atgtctagac tggacaagag caaagtcata
     4321 aacggagctc tggaattact caatggtgtc ggtatcgaag gcctgacgac aaggaaactc
     4381 gctcaaaagc tgggagttga gcagcctacc ctgtactggc acgtgaagaa caagcgggcc
     4441 ctgctcgatg ccctgccaat cgagatgctg gacaggcatc atacccactt ctgccccctg
     4501 gaaggcgagt catggcaaga ctttctgcgg aacaacgcca agtcataccg ctgtgctctc
     4561 ctctcacatc gcgacggggc taaagtgcat ctcggcaccc gcccaacaga gaaacagtac
     4621 gaaaccctgg aaaatcagct cgcgttcctg tgtcagcaag gcttctccct ggagaacgca
     4681 ctgtacgctc tgtccgccgt gggccacttt acactgggct gcgtattgga ggaacaggag
     4741 catcaagtag caaaagagga aagagagaca cctaccaccg attctatgcc cccacttctg
     4801 agacaagcaa ttgagctgtt cgaccggcag ggagccgaac ctgccttcct tttcggcctg
     4861 gaactaatca tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg gccgaccgac
     4921 gcccttgacg attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt
     4981 gatatgctgc ctgctgacgc tcttgacgat tttgaccttg acatgctccc cgggtaacta
     5041 agtaaggatc gatccaagat atcgtattct taactatgtt gctcctttta cgctatgtgg
     5101 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     5161 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     5221 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     5281 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     5341 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     5401 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     5461 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     5521 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     5581 gagtcggatc tccctttggg ccgcctcccc gcctgtttcg cctcggcgtc cggactagag
     5641 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     5701 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     5761 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     5821 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     5881 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     5941 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     6001 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6061 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6121 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc
     6181 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6241 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     6301 tttttggagg cctttcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     6361 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     6421 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     6481 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     6541 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     6601 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     6661 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6721 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6781 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     6841 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     6901 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     6961 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     7021 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     7081 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     7141 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     7201 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     7261 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     7321 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     7381 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     7441 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     7501 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     7561 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     7621 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7681 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7741 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     7801 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     7861 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     7921 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7981 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     8041 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     8101 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     8161 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     8221 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     8281 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     8341 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     8401 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
     8461 tgcaagctta atgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag
     8521 ttagcaacat gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag
     8581 gtggtacgat cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac
     8641 cactgaattg ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg
     8701 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     8761 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     8821 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     8881 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
     8941 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
     9001 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
     9061 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
     9121 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
     9181 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
     9241 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
     9301 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
     9361 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga
     9421 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     9481 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     9541 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     9601 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     9661 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     9721 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     9781 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     9841 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     9901 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     9961 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
    10021 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
    10081 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
    10141 attatcgttt cagacccacc tcccaacccc gaggggac
//
