LOCUS       sequence-386565-       10696 bp    DNA     circular SYN 14-APR-2025
DEFINITION  LV construct encoding GFP-TMD-DLL1 ligand under dox-inducible TRE 
            promoter and containing a PGK-rtTA-M2-IRES-Puro regulator/resistance
            marker cassette..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10696)
  AUTHORS   Sloas DC, Tran JC, Marzilli AM, Ngo JT
  TITLE     Tension-tuned receptors for synthetic mechanotransduction and 
            intercellular force detection.
  JOURNAL   Nat Biotechnol. 2023 Jan 16. doi: 10.1038/s41587-022-01638-y.
  PUBMED    36646932
REFERENCE   2  (bases 1 to 10696)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2023 Jan 16. doi: 10.1038/s41587-022-01638-y."
COMMENT     Sequence Label: LV TRE-GFP-TMD-DLL1
FEATURES             Location/Qualifiers
     source          1..10696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             32..212
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    259..384
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    877..1110
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1295..1339
                     /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    1606..1723
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    join(1756..1774,1775..1797,1798..1816,1817..1839,1840..1858,
                     1859..1881,1882..1900,1901..1923,1924..1942,1943..1965,
                     1966..1984,1985..2007,2008..2026)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2059..2097
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2094..2118
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2270..2279
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2339..2341,2342..2344,2345..3055)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        3780..4290
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     4209..4227
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             join(4328..4945,4946..4951,4952..4987,4988..4990,4991..5026,
                     5027..5029,5030..5065,5066..5074)
                     /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"
     misc_feature    join(5119..5693,5694..5696,5697..5705)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5286..5303)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5513..5532
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5706..6305
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5706..5725)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6202..6222
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6365..6953
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6418..6438)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6836..6847)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7040..7273
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7345..7466
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7382..7401)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7436..7455
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7485..7620
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7640..7659)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7641..7659)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7669..7686)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7669..7685)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7678..7700)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7827..8282
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7914..7933)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8124..8145
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8308..8412
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8413..8481,8482..9273)
                     /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(8631..8650)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9444..10032
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9933..9952
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10186..10203
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10320..10341
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10356..10361,10362..10379,10380..10386)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10394..10410
                     /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     10399..10421
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10418..10434
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10418..10434
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10453..10473
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10455..10473
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        join(10501..10696,1..31)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 tgtatttaag tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc
       61 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
      121 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
      181 acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg
      241 aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca
      301 agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag
      361 gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa
      421 aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca
      481 agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt
      541 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca
      601 ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc
      661 aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa
      721 gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
      781 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
      841 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
      901 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
      961 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
     1021 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
     1081 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
     1141 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
     1201 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
     1261 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
     1321 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
     1381 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
     1441 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
     1501 cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga
     1561 cagatccatt cgattagtga acggatctcg acggtatcgg ttaactttta aaagaaaagg
     1621 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     1681 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttatcgatc acgagactag
     1741 cctcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
     1801 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
     1861 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
     1921 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata
     1981 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
     2041 gctcggtacc cgggtcgagg taggcgtgta cggtgggagg cctatataag cagagctcgt
     2101 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
     2161 caccgggacc gatccagcct ccgcggcccc gaattcgagc tcggtacccg gggatcctct
     2221 agtcagctga cgcgtcaagt ggagcaaggc aggtggacag tggatcgccg ccaccatgga
     2281 gaccgacacc ctcttgctct gggtcctgct gctgtgggtt ccgggtagta ccggagacat
     2341 ggtttctaag ggtgaagaat tgttcaccgg agttgtgccc atcctggttg agctggacgg
     2401 agacgttaac ggacacaagt tctcagtgag cggcgaagga gagggcgacg ctacgtacgg
     2461 taaactgacg ttgaagttca tctgtacaac aggaaagctg ccagtgccct ggcctacact
     2521 cgtgaccacc ctcacctacg gagtccagtg cttctcccgg taccccgacc acatgaaaca
     2581 gcacgatttc ttcaagtccg cgatgcctga gggctatgtt caggaacgca ccatattctt
     2641 caaggacgac ggtaactata aaaccagggc tgaagttaag ttcgaggggg acacgttggt
     2701 aaatagaatt gagttgaaag gaatcgattt caaagaagac ggcaatattt tgggccacaa
     2761 gctggagtac aactacaaca gccacaacgt gtatattatg gctgataagc agaagaacgg
     2821 catcaaggtg aacttcaaaa tccgacacaa tatcgaggat ggctctgtgc aactggccga
     2881 ccattaccag caaaacacgc ctatcggcga cggtccggtt ttgctgccag ataaccacta
     2941 cctctccacg caatcagccc tctccaagga tcctaacgaa aaaagagacc atatggttct
     3001 gcttgaattc gtgacggctg ctgggatcac cctgggaatg gatgagctgt ataagaacgc
     3061 cgtgggccag gatacgcagg aggtgatcgt cgtgccccac agcttgccct tcaaggtcgt
     3121 ggttattagt gctatcctgg cgttggtggt gctgactatt atctccctta tcatacttat
     3181 catgctgtgg caaaaaaaac cccgagtccg gctgaagcta cagaaacacc agcctccgcc
     3241 tgatccttgc gggggagaga cagagaccat gaacaaccta gccaattgcc agcgtgagaa
     3301 ggatgtttct gttagcatca ttggggctac acagatcaag aacaccaaca agaaggcgga
     3361 ctttcatggg gaccatggtg ctgacaagag cagctttaag gcccgatacc ccactgtgga
     3421 ctataacctc attcgagacc tcaagggaga tgaagccacg gtcagggatg cacacagcaa
     3481 acgtgacacc aagtgccagt cacagggctc tgtaggagaa gagaagagca cctcaacgct
     3541 caggggtggg gaggttcccg acagaaaaag gcccgagtct gtctactcta cttcaaagga
     3601 caccaagtac cagtcggtgt atgttctatc tgcagaaaag gatgagtgtg ttatagcgac
     3661 tgaggtttaa gatccttgac ttgcggccgc aactcccacc tgcaacatgc gtgactgact
     3721 gaggccgcga ctctagcgcg gccgcatcga taagcttgtg gacgatatcg aattcgcacg
     3781 gggttggggt tgcgcctttt ccaaggcagc cctgggtttg cgcagggacg cggctgctct
     3841 gggcgtggtt ccgggaaacg cagcggcgcc gaccctgggt ctcgcacatt cttcacgtcc
     3901 gttcgcagcg tcacccggat cttcgccgct acccttgtgg gccccccggc gacgcttcct
     3961 gctccgcccc taagtcggga aggttccttg cggttcgcgg cgtgccggac gtgacaaacg
     4021 gaagccgcac gtctcactag taccctcgca gacggacagc gccagggagc aatggcagcg
     4081 cgccgaccgc gatgggctgt ggccaatagc ggctgctcag cagggcgcgc cgagagcagc
     4141 ggccgggaag gggcggtgcg ggaggcgggg tgtggggcgg tagtgtgggc cctgttcctg
     4201 cccgcgcggt gttccgcatt ctgcaagcct ccggagcgca cgtcggcagt cggctccctc
     4261 gttgaccgaa tcaccgacct ctctccccag ggggatccgt ttatctgcag aattcgccct
     4321 tgacgtcatg tctagactgg acaagagcaa agtcataaac ggcgctctgg aattactcaa
     4381 tggagtcggt atcgaaggcc tgacgacaag gaaactcgct caaaagctgg gagttgagca
     4441 gcctaccctg tactggcacg tgaagaacaa gcgggccctg ctcgatgccc tgccaatcga
     4501 gatgctggac aggcatcata cccacttctg ccccctggaa ggcgagtcat ggcaagactt
     4561 tctgcggaac aacgccaagt cattccgctg tgctctcctc tcacatcgcg acggggctaa
     4621 agtgcatctc ggcacccgcc caacagagaa acagtacgaa accctggaaa atcagctcgc
     4681 gttcctgtgt cagcaaggct tctccctgga gaacgcactg tacgctctgt ccgccgtggg
     4741 ccactttaca ctgggctgcg tattggagga acaggagcat caagtagcaa aagaggaaag
     4801 agagacacct accaccgatt ctatgccccc acttctgaga caagcaattg agctgttcga
     4861 ccggcaggga gccgaacctg ccttcctttt cggcctggaa ctaatcatat gtggcctgga
     4921 gaaacagcta aagtgcgaaa gcggcgggcc ggccgacgcc cttgacgatt ttgacttaga
     4981 catgctccca gccgatgccc ttgacgactt tgaccttgat atgctgcctg ctgacgctct
     5041 tgacgatttt gaccttgaca tgctccccgg gtaacggacc ggacgtcaag ggcgaattcc
     5101 agcacactgg gaattccgcc cctctccctc ccccccccct aacgttactg gccgaagccg
     5161 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
     5221 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
     5281 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
     5341 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
     5401 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
     5461 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
     5521 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
     5581 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
     5641 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca
     5701 caaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     5761 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     5821 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     5881 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     5941 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     6001 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     6061 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     6121 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     6181 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     6241 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     6301 cctgaccagc acactggcgg ccgttactag cttctgcagc acgtgtctag aaacgctggt
     6361 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     6421 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     6481 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     6541 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     6601 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     6661 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     6721 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     6781 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     6841 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     6901 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat
     6961 tcgagctcgg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     7021 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
     7081 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     7141 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     7201 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     7261 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     7321 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
     7381 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     7441 ttgtccaaac tcatcaatgt atcttatcat gtctggctct ascyatcccg cccctaactc
     7501 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     7561 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     7621 tagggacgta cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt
     7681 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     7741 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     7801 gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     7861 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     7921 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     7981 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     8041 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     8101 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     8161 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     8221 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     8281 ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     8341 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     8401 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     8461 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     8521 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     8581 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     8641 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     8701 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     8761 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     8821 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     8881 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     8941 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     9001 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     9061 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     9121 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     9181 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     9241 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     9301 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     9361 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     9421 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     9481 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     9541 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     9601 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     9661 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     9721 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     9781 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     9841 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     9901 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     9961 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
    10021 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
    10081 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
    10141 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
    10201 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
    10261 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
    10321 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
    10381 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
    10441 tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctggagct gcaagcttaa
    10501 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
    10561 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
    10621 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
    10681 cgcattgcag agatat
//
