LOCUS       sequence-474331-       10106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Blasticidin selected lentiviral vector with Tet-On regulatory 
            proteins and the cumate repressor, for use with dual-inducible 
            CymR-CuO system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10106)
  AUTHORS   Kent MR, Jay AN, Kendall GC
  TITLE     New dual inducible cellular model to investigate temporal control of
            oncogenic cooperating genes.
  JOURNAL   Sci Rep. 2024 Sep 5;14(1):20773. doi: 10.1038/s41598-024-71227-3.
  PUBMED    39237585
REFERENCE   2  (bases 1 to 10106)
  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: "Sci Rep."; 
            date: "2024-09-5"; volume: "14(1):20773. doi"; pages: " 
            10.1038/s41598-024-71227-3"
COMMENT     Sequence Label: pLV-Bsd-CMV>tTS/rtTA:T2A:{cymR}
FEATURES             Location/Qualifiers
     source          1..10106
                     /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"
     enhancer        2019..2322
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2323..2526
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2477..2497
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2547..2571
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(2578..3210,3211..3231,3232..3423)
                     /codon_start=1
                     /product="fusion of a hybrid tetracycline repressor TetR 
                     with a nuclear localization signal and a KRAB silencing 
                     domain"
                     /label=tTS
                     /note="silences Tet-responsive promoters in the absence but
                     not the presence of doxycycline"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     RNKQTLMNMLSEAILAKHHTRSAPLPTESWQQFLQENALSFRKALLVHRDGARLHIGTS
                     PTPPQFEQAEAQLRCLCDAGFSVEEALFILQSISHFTLGAVLEEQATNQIENNHVIDAA
                     PPLLQEAFNIQARTSAEMAFHFGLKSLIFGFSAQLDEKKHTPIEDGNKPKKKRKLAVSV
                     TFEDVAVLFTRDEWKKLDLSQRSLYREVMLENYSNLASMAGFLFTKPKVISLLQQGEDP
                     W"
     primer_bind     complement(2651..2670)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             3433..3486
                     /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(3487..4104,4105..4110,4111..4146,4147..4149,4150..4185,
                     4186..4188,4189..4224,4225..4230)
                     /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
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             4240..4293
                     /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             4294..4905
                     /codon_start=1
                     /gene="cymR from Pseudomonas putida"
                     /product="cumate repressor (Mullick et al., 2006)"
                     /label=CymR
                     /translation="MSPKRRTQAERAMETQGKLIAAALGVLREKGYAGFRIADVPGAAG
                     VSRGAQSHHFPTKLELLLATFEWLYEQITERSRARLAKLKPEDDVIQQMLDDAAEFFLD
                     DDFSISLDLIVAADRDPALREGIQRTVERNRFVVEDMWLGVLVSRGLSRDDAEDILWLI
                     FNSVRGLAVRSLWQKDKERFERVRNSTLEIARERYAKFKR"
     protein_bind    complement(4906..4930)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4948..5536
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5001..5021)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5419..5430)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        5567..6066
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5602..5625)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5999..6018
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        6076..6123
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6142..6540
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             6627..6860
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6932..7053
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6969..6988)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7023..7042
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7093..7228
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7248..7267)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7249..7267)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7277..7294)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7277..7293)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7286..7308)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7435..7890
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7522..7541)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7732..7753
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7916..8020
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8021..8089,8090..8881)
                     /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(8239..8258)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9052..9640
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9541..9560
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9794..9811
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9928..9949
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9964..9969,9970..9987,9988..9994)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10002..10018
                     /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     10007..10029
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10026..10042
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10026..10042
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10061..10081
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10063..10081
                     /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 aaaagttgta gttattaata gtaatcaatt
     1981 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     2041 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     2101 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     2161 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     2221 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     2281 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     2341 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     2401 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     2461 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     2521 agagctggtt tagtgaaccg tcagatccaa gtttgtacaa aaaagcaggc tgccaccatg
     2581 tctagattag ataaaagtaa agtgattaac agcgcattag agctgcttaa tgaggtcgga
     2641 atcgaaggtt taacaacccg taaactcgcc cagaagctag gtgtagagca gcctacattg
     2701 tattggcacg tgcgcaacaa gcagactctt atgaacatgc tttcagaggc aatactggcg
     2761 aagcatcaca cccgttcagc accgttaccg actgagagtt ggcagcagtt tctccaggaa
     2821 aatgctctga gtttccgtaa agcattactg gtccatcgtg atggagcccg attgcatata
     2881 gggacctctc ctacgccccc ccagtttgaa caagcagagg cgcaactacg ctgtctatgc
     2941 gatgcagggt tttcggtcga ggaggctctt ttcattctgc aatctatcag ccattttacg
     3001 ttgggtgcag tattagagga gcaagcaaca aaccagatag aaaataatca tgtgatagac
     3061 gctgcaccac cattattaca agaggcattt aatattcagg cgagaacctc tgctgaaatg
     3121 gccttccatt tcgggctgaa atcattaata tttggatttt ctgcacagtt agatgaaaaa
     3181 aagcatacac ccattgagga tggtaataaa ccaaaaaaga agagaaagct agcagtgtca
     3241 gtgacatttg aagatgtggc tgtgctcttt actcgggacg agtggaagaa gctggatctg
     3301 tctcagagaa gcctgtaccg tgaggtgatg ctggagaatt acagcaacct ggcctccatg
     3361 gcaggattcc tgtttaccaa accaaaggtg atctccctgt tgcagcaagg agaggatccc
     3421 tggggctccg gagagggcag gggaagtctt ctaacatgcg gggacgtgga ggaaaatccc
     3481 ggccccatgt ctagactgga caagagcaaa gtcataaacg gcgctctgga attactcaat
     3541 ggagtcggta tcgaaggcct gacgacaagg aaactcgctc aaaagctggg agttgagcag
     3601 cctaccctgt actggcacgt gaagaacaag cgggccctgc tcgatgccct gccaatcgag
     3661 atgctggaca ggcatcatac ccacttctgc cccctggaag gcgagtcatg gcaagacttt
     3721 ctgcggaaca acgccaagtc attccgctgt gctctcctct cacatcgcga cggggctaaa
     3781 gtgcatctcg gcacccgccc aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg
     3841 ttcctgtgtc agcaaggctt ctccctggag aacgcactgt acgctctgtc cgccgtgggc
     3901 cactttacac tgggctgcgt attggaggaa caggagcatc aagtagcaaa agaggaaaga
     3961 gagacaccta ccaccgattc tatgccccca cttctgagac aagcaattga gctgttcgac
     4021 cggcagggag ccgaacctgc cttccttttc ggcctggaac taatcatatg tggcctggag
     4081 aaacagctaa agtgcgaaag cggcgggccg gccgacgccc ttgacgattt tgacttagac
     4141 atgctcccag ccgatgccct tgacgacttt gaccttgata tgctgcctgc tgacgctctt
     4201 gacgattttg accttgacat gctccccggg ggaagcggag agggcagggg aagtcttcta
     4261 acatgcgggg acgtggagga aaatcccggc cccatgtcgc cgaagcgccg cacccaggcc
     4321 gagcgcgcga tggaaaccca gggcaagctg atcgccgccg ccctgggcgt cctgcgcgag
     4381 aagggctatg ccggcttccg catcgccgac gtcccgggcg ccgccggcgt cagccgcggc
     4441 gcccagtcgc accacttccc gaccaagctg gaactgctgc tggccacctt cgagtggctg
     4501 tacgagcaga tcaccgagcg cagccgcgcc cgcctggcca agctgaagcc cgaggacgac
     4561 gtcatccagc agatgctgga cgacgccgcc gagttcttcc tggacgacga cttctcgatc
     4621 tcgctggacc tgatcgtggc cgccgaccgc gacccggcgc tgcgcgaggg catccagcgc
     4681 accgtcgagc gcaaccgctt cgtcgtcgag gatatgtggc tgggcgtgct ggtgtcgcgc
     4741 ggcctgagcc gcgacgacgc ggaggacatc ctgtggctga tcttcaactc ggtccgcggc
     4801 ctggcggtcc gctcgctgtg gcagaaggac aaggagcgct tcgagcgcgt ccgcaactcg
     4861 accctggaaa tcgcccgcga gcgctacgcc aagttcaagc gctgaaccca gctttcttgt
     4921 acaaagtggt gataatcgaa ttccgataat caacctctgg attacaaaat ttgtgaaaga
     4981 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     5041 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     5101 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     5161 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     5221 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     5281 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     5341 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     5401 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     5461 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     5521 tgggccgcct ccccgcatcg ggaattcccg cggttcgaat tctaccgggt aggggaggcg
     5581 cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac
     5641 acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg
     5701 ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc
     5761 gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc
     5821 gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa
     5881 tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc
     5941 gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca
     6001 ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc
     6061 ctttcgacct cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac
     6121 gacaaggtga ggaactaaac catggccaag cctttgtctc aagaagaatc caccctcatt
     6181 gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag cgtcgccagc
     6241 gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca ttttactggg
     6301 ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc tggcaacctg
     6361 acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg cggacggtgc
     6421 cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga cagtgatgga
     6481 cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa
     6541 gcacaattcg agctcggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     6601 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     6661 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     6721 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     6781 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     6841 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     6901 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     6961 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     7021 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     7081 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     7141 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     7201 aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt
     7261 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     7321 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     7381 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     7441 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     7501 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     7561 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     7621 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     7681 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     7741 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     7801 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     7861 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     7921 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     7981 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     8041 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     8101 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     8161 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     8221 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     8281 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     8341 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     8401 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     8461 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     8521 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     8581 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     8641 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     8701 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     8761 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     8821 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     8881 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     8941 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     9001 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     9061 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     9121 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     9181 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     9241 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     9301 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     9361 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     9421 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag agagaaaggc
     9481 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     9541 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     9601 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     9661 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     9721 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     9781 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     9841 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     9901 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     9961 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
    10021 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
    10081 ggaacaaaag ctggagctgc aagctt
//
