LOCUS       sequence-387892-       10508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TET-On reverse transactivator labeled with Antares.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10508)
  AUTHORS   Hurt RC, Buss MT, Duan M, Wong K, You MY, Sawyer DP, Swift MB, Dutka
            P, Barturen-Larrea P, Mittelstein DR, Jin Z, Abedi MH, Farhadi A, 
            Deshpande R, Shapiro MG
  TITLE     Genomically mined acoustic reporter genes for real-time in vivo 
            monitoring of tumors and tumor-homing bacteria.
  JOURNAL   Nat Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01581-y.
  PUBMED    36593411
REFERENCE   2  (bases 1 to 10508)
  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: "Nat 
            Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01581-y."
COMMENT     Sequence Label: pEF1alpha-rtTA-Antares
FEATURES             Location/Qualifiers
     source          1..10508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      540..1128
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1029..1048
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1282..1299
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1416..1437
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1452..1457,1458..1475,1476..1482)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1490..1506
                     /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     1495..1517
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1514..1530
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1514..1530
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1549..1569
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1551..1569
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        1670..2049
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2050..2253
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2204..2224
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2268..2448
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2495..2620
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3113..3346
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3531..3575
                     /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    3868..3985
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4033..5211
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4264..5202
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5159..5179
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             5217..5960
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             5976..6032
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             6714..7220
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="FTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRIV
                     LSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVTP
                     NMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWRL
                     CERILA"
     misc_feature    7923..8511
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7976..7996)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8394..8405)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8623..8856
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8928..9049
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8965..8984)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9019..9038
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9089..9224
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9244..9263)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9245..9263)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9273..9290)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9273..9289)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9282..9304)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9431..9886
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9518..9537)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9728..9749
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9912..10016
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10017..10085,10086..10508,1..369)
                     /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(10235..10254)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
       61 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
      121 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
      181 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
      241 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
      301 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
      361 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
      421 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
      481 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
      541 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
      601 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
      661 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
      721 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
      781 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
      841 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
      901 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
      961 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     1021 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     1081 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     1141 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     1201 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     1261 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     1321 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     1381 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     1441 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     1501 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct
     1561 cactaaaggg aacaaaagct ggagctgcaa gcttggccat tgcatacgtt gtatccatat
     1621 cataatatgt acatttatat tggctcatgt ccaacattac cgccatgttg acattgatta
     1681 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     1741 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     1801 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     1861 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     1921 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     1981 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     2041 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     2101 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     2161 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     2221 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgggg tctctctggt
     2281 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     2341 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     2401 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
     2461 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
     2521 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
     2581 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
     2641 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
     2701 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
     2761 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
     2821 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
     2881 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
     2941 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
     3001 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     3061 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     3121 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     3181 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     3241 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     3301 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttagggtt
     3361 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     3421 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     3481 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     3541 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     3601 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     3661 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     3721 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     3781 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgatctc
     3841 gacacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag
     3901 tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag aattacaaaa
     3961 acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg
     4021 gggtacagaa ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     4081 gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta
     4141 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     4201 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     4261 caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc
     4321 gtgccttgaa ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt
     4381 ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga
     4441 gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc
     4501 tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg
     4561 ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg
     4621 gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg
     4681 cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct
     4741 gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc
     4801 cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc
     4861 tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa
     4921 agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc
     4981 aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg
     5041 ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg
     5101 cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc
     5161 aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg acacggatgt
     5221 ctagactgga caagagcaaa gtcataaact ctgctctgga attactcaat ggagtcggta
     5281 tcgaaggcct gacgacaagg aaactcgctc aaaagctggg agttgagcag cctaccctgt
     5341 actggcacgt gaagaacaag cgggccctgc tcgatgccct gccaatcgag atgctggaca
     5401 ggcatcatac ccactcctgc cccctggaag gcgagtcatg gcaagacttt ctgcggaaca
     5461 acgccaagtc ataccgctgt gctctcctct cacatcgcga cggggctaaa gtgcatctcg
     5521 gcacccgccc aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg ttcctgtgtc
     5581 agcaaggctt ctccctggag aacgcactgt acgctctgtc cgccgtgggc cactttacac
     5641 tgggctgcgt attggaggaa caggagcatc aagtagcaaa agaggaaaga gagacaccta
     5701 ccaccgattc tatgccccca cttctgaaac aagcaattga gctgttcgac cggcagggag
     5761 ccgaacctgc cttccttttc ggcctggaac taatcatatg tggcctggag aaacagctaa
     5821 agtgcgaaag cggcgggccg accgacgccc ttgacgattt tgacttagac atgctcccag
     5881 ccgatgccct tgacgacttt gaccttgata tgctgcctgc tgacgctctt gacgattttg
     5941 accttgacat gctccccggg ggatccggaa gcggagctac taacttcagc ctgctgaagc
     6001 aggctggaga cgtggaggag aaccctggac ctgtgagcaa gggcgaggag ctgatcaagg
     6061 agaacatgag aagcaagctg tacctggaag gcagcgtgaa cggccaccag ttcaagtgca
     6121 cccacgaagg ggagggcaag ccctacgagg gcaagcagac caacaggatc aaggtggtgg
     6181 agggaggccc cctgccgttc gcattcgaca tcctggccac ccactttatg tacgggagca
     6241 aggtgttcat caagtacccc gccgacctcc ccgattattt taagcagtcc ttccctgagg
     6301 gcttcacatg ggagagagtc atggtgttcg aagacggggg cgtgctgacc gccacccagg
     6361 acaccagcct ccaggacggc gagctcatct acaacgtcaa ggtcagaggg gtgaacttcc
     6421 cagccaacgg ccccgtgatg cagaagaaaa cactgggctg ggagcccagc accgagacca
     6481 tgtaccccgc tgacggcggc ctggaaggca gatgcgacaa ggccctgaag ctcgtgggcg
     6541 ggggccacct gcacgtcaac ttcaagacca catacaagtc caagaaaccc gtgaagatgc
     6601 ccggcgtcca ctacgtggac cgcagactgg aaagaatcaa ggaggccgac aacgagacct
     6661 acgtcgagca gtacgagcac gctgtggcca gatactccaa cctgggcgga ggcttcacac
     6721 tcgaagattt cgttggggac tggcgacaga cagccggcta caacctggac caagtccttg
     6781 aacagggagg tgtgtccagt ttgtttcaga atctcggggt gtccgtaact ccgatccaaa
     6841 ggattgtcct gagcggtgaa aatgggctga agatcgacat ccatgtcatc atcccgtatg
     6901 aaggtctgag cggcgaccaa atgggccaga tcgaaaaaat ttttaaggtg gtgtaccctg
     6961 tggatgatca tcactttaag gtgatcctgc actatggcac actggtaatc gacggggtta
     7021 cgccgaacat gatcgactat ttcggacggc cgtatgaagg catcgccgtg ttcgacggca
     7081 aaaagatcac tgtaacaggg accctgtgga acggcaacaa aattatcgac gagcgcctga
     7141 tcaaccccga cggctccctg ctgttccgag taaccatcaa cggagtgacc ggctggcggc
     7201 tgtgcgaacg cattctggcg aggcacgagc tgatcaagga gaacatgaga agcaagctgt
     7261 acctggaagg cagcgtgaac ggccaccagt tcaagtgcac ccacgaaggg gagggcaagc
     7321 cctacgaggg caagcagacc aacaggatca aggtggtgga gggaggcccc ctgccgttcg
     7381 cattcgacat cctggccacc cactttatgt acgggagcaa ggtgttcatc aagtaccccg
     7441 ccgacctccc cgattatttt aagcagtcct tccctgaggg cttcacatgg gagagagtca
     7501 tggtgttcga agacgggggc gtgctgaccg ccacccagga caccagcctc caggacggcg
     7561 agctcatcta caacgtcaag gtcagagggg tgaacttccc agccaacggc cccgtgatgc
     7621 agaagaaaac actgggctgg gagcccagca ccgagaccat gtaccccgct gacggcggcc
     7681 tggaaggcag atgcgacaag gccctgaagc tcgtgggcgg gggccacctg cacgtcaact
     7741 tcaagaccac atacaagtcc aagaaacccg tgaagatgcc cggcgtccac tacgtggacc
     7801 gcagactgga aagaatcaag gaggccgaca acgagaccta cgtcgagcag tacgagcacg
     7861 ctgtggccag atactccaac ctgggcggag gcatggacga gctgtacaag gggtaatgta
     7921 caaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     7981 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     8041 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     8101 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     8161 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     8221 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     8281 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     8341 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     8401 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     8461 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg ctacaaggca
     8521 gctgtagatc cctttcatag aaggcggcgg tggtaccttt aagaccaatg acttacaagg
     8581 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     8641 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     8701 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     8761 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     8821 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     8881 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     8941 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     9001 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     9061 tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     9121 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     9181 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat
     9241 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
     9301 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     9361 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     9421 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     9481 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     9541 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     9601 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     9661 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     9721 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     9781 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     9841 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     9901 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     9961 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
    10021 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
    10081 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
    10141 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
    10201 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
    10261 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
    10321 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
    10381 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
    10441 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
    10501 gttgggaa
//
