LOCUS       sequence-340012-       11079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian acoustic reporter gene 1 cassette 2 on XLone plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11079)
  AUTHORS   Howells AR, Welch PJ, Kim J, Forest CR, Shi C, Lian XL
  TITLE     A drug-selectable acoustic reporter gene system for human cell 
            ultrasound imaging.
  JOURNAL   Bioeng Transl Med. 2023 Aug 2;9(2):e10584. doi: 10.1002/btm2.10584. 
            eCollection 2024 Mar.
  PUBMED    38435822
REFERENCE   2  (bases 1 to 11079)
  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: "Bioeng 
            Transl Med."; date: "2023-08-2"; volume: "9(2):e10584. doi"; pages: 
            " 10.1002/btm2.10584. eCollection 2024 Mar"
COMMENT     Sequence Label: XLone-mARG2
FEATURES             Location/Qualifiers
     source          1..11079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        218..585
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    226..244
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    262..280
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    298..316
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    334..352
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    370..388
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    406..424
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    442..460
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1540..1596
                     /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"
     regulatory      1723..1732
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1727..1736
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2380..2436
                     /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             2719..2775
                     /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             3601..3657
                     /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             3961..4017
                     /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             4318..4374
                     /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             4693..4749
                     /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             5182..5238
                     /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"
     primer_bind     complement(5545..5564)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    6139..6273
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   6549..6583
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6594..6611)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6594..6610)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6603..6625)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6819..6838)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6938..6960
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6998..7016)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7084..7188
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7189..7257,7258..8049)
                     /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(7407..7426)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8220..8808
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8709..8728
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8962..8979
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9096..9117
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9132..9137,9138..9155,9156..9162)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9170..9186
                     /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     9175..9197
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9194..9210
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9194..9210
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   9236..9298
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    9474..9555
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(9576..10175)
                     /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(9659..9679)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     10156..10175
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(10185..10238)
                     /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             complement(10248..10991)
                     /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             complement(10992..11015)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      11015..11024
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(join(11044..11079,1..176))
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
ORIGIN
        1 gttcacggcg actactgcac ttatatacgg ttctccccca ccctcgggaa aaaggcggag
       61 ccagtacacg acatcacttt cccagtttac cccgcgccac cttctctagg caccggttca
      121 attgccgacc cctcccccca acttctcggg gactgtgggc gatgtgcgct ctgcccactg
      181 acgggcaccg gagccactcg agtggaatta tcacctcgag tttactccct atcagtgata
      241 gagaacgtat gaagagttta ctccctatca gtgatagaga acgtatgcag actttactcc
      301 ctatcagtga tagagaacgt ataaggagtt tactccctat cagtgataga gaacgtatga
      361 ccagtttact ccctatcagt gatagagaac gtatctacag tttactccct atcagtgata
      421 gagaacgtat atccagttta ctccctatca gtgatagaga acgtataagc tttgcttatg
      481 taaaccaggg cgcctataaa agagtgctga ttttttgagt aaacttcaat tccacaacac
      541 ttttgtctta taccaacttt ccgtaccact tcctaccctc gtaaaggtac cgccaccatg
      601 accgtgctga ccgacaagcg gaagaagggc agcggcgcct tcatccagga cgacgagaca
      661 aaagaggtgc tgagcagagc cctgagctac ctgaagtccg gctacagcat ccacttcacc
      721 ggacctgccg gcggaggcaa gacatctctg gctagagccc tggccaagaa acggaagcgg
      781 cccgtgatgc tgatgcacgg caaccacgag ctgaacaaca aggacctgat cggcgatttc
      841 accggctaca ccagcaaaaa ggtgatcgac cagtacgtgc ggagcgtgta caagaaagac
      901 gaacaggtgt ccgagaactg gcaggacggc agactgctgg aagccgtgaa gaatggctac
      961 accctgatct acgacgagtt caccagaagc aagcccgcta ccaacaacat cttcctgagc
     1021 atccttgagg agggcgtgct gcccctgtac ggcgtgaaga tgaccgaccc tttcgtgcgc
     1081 gtgcaccccg acttcagagt gatctttacc agcaaccccg ccgagtatgc cggcgtgtac
     1141 gatacccagg acgccctgct ggaccggctg atcaccatgt tcatcgacta caaggacatc
     1201 gaccgggaaa ccgctatcct gaccgagaaa actgacgtgg aagaagacga ggcccggacc
     1261 atcgtgaccc tggtggccaa cgtgcggaac agaagcggcg acgagaatag cagcggcctg
     1321 agcctgagag ccagcctgat gattgccacc ctggccaccc agcaggacat ccctatcgat
     1381 ggcagcgacg aggacttcca gaccctgtgc atcgacatcc tgcaccaccc cctgaccaag
     1441 tgcctggacg aagagaacgc caagagcaag gccgagaaga tcattctcga agagtgcaag
     1501 aacatcgaca ccgaggagaa gggtgccccg ggatctggcg caacaaattt tagtctttta
     1561 aagcaggcag gagacgtcga ggaaaaccct ggacccgtga gcgagacaaa cgagacaggc
     1621 atctacatct tcagcgccat ccagacagac aaggatgagg aattcggcgc cgtggaagtg
     1681 gaagggacca aggctgagac attcctgatc cggtataagg acgccgccat ggtggccgcc
     1741 gaagtgccca tgaagatcta ccaccccaac cggcagaacc tgctgatgca ccagaatgcc
     1801 gtggccgcca tcatggacaa gaacgacacc gtgatcccca tcagcttcgg caacgtgttc
     1861 aagagcaaag aggacgtgaa ggtgctcctg gaaaacctgt acccccagtt cgagaagctg
     1921 ttccccgcca tcaagggaaa gatcgaagtg ggcctgaagg tgatcggcaa gaaagagtgg
     1981 ctcgaaaaga aagtgaacga gaaccccgag ctggaaaaag tgtccgccag cgtgaagggc
     2041 aagagcgagg ccgctggcta ctacgagaga atccagctgg gcggcatggc ccagaagatg
     2101 ttcacaagcc tgcagaaaga agtgaaaacc gacgtgttca gccccctgga agaagccgcc
     2161 gaggccgcca aagccaatga gcctacaggc gaaacaatgc tgctgaacgc cagcttcctg
     2221 atcaacagag aggatgaggc caagttcgac gagaaagtca atgaggccca cgagaactgg
     2281 aaggataagg ccgacttcca ctacagcggc ccctggcccg cctacaactt cgtgaacatc
     2341 cggctgaagg tggaagagaa gggggcacct ggctcgggag cgaccaactt ctcattactc
     2401 aaacaagccg gagacgttga ggagaatcca ggccctgtgc tgcacaagct cgtgaccgcc
     2461 cccatcaacc tggtcgtgaa gatcggcgag aaggtgcagg aagaggccga caagcagctg
     2521 tacgacctgc ccaccatcca gcagaagctg atccagctgc agatgatgtt cgagctgggc
     2581 gagatccccg aggaagcctt ccaggaaaaa gaggacgaac tgctgatgag atacgagatc
     2641 gccaagcggc gcgagattga gcagtgggaa gaactgaccc agaagcggaa tgaggaaagc
     2701 ggtgccccgg gatctggcgc aacaaatttt agtcttttaa agcaggcagg agacgtcgag
     2761 gaaaaccctg gacccgtggg cgagctgctg tacctctacg gcctgatccc caccaaagag
     2821 gccgctgcta tcgagccctt cccattctac aagggcttcg acggcgagca cagcctgtac
     2881 cctatcgcct tcgaccaagt gaccgccgtg gtgttcaagc tggacgccga cacctacagc
     2941 gagaaagtga tccaggaaaa gatggaacag gacatgagct ggctgcagga aaaggccttc
     3001 caccaccacg agacagtggc cgccctgtat gaggaattca ccatcatccc cctgaagttc
     3061 tgcaccatct ataagggaga ggaatccctg caggccgcca tcgagatcaa caaagagaag
     3121 atcgaaaact ccctgaccct gctgcagggc aacgaggaat ggaacgtgaa gatctactgc
     3181 gacgacaccg agctgaagaa gggcatcagc gagacaaacg agagcgtgaa ggccaagaag
     3241 caggaaatca gccacctgag ccccggcaga cagttcttcg agaagaagaa gattgaccag
     3301 ctcatcgaga aagagctgga actgcacaag aacaaagtgt gcgaggaaat ccacgacaag
     3361 ctgattgagc tgagcctcta cgactccgtg aagaagaact ggtccaagga cgtgacaggc
     3421 gctgccgaac agatggcctg gaacagcgtg ttcctgctgc ccagcctgca gatcaccaag
     3481 ttcgtgaacg agatcgagga actccagcag cggctggaga acaagggatg gaagttcgaa
     3541 gtgaccggcc cctggcctcc ctaccacttc agcagctttg ccggggcacc tggctcggga
     3601 gcgaccaact tctcattact caaacaagcc ggagacgttg aggagaatcc aggccctgtg
     3661 agcctgaagc agagcatgga gaataaggat atcgccctga tcgacatcct cgacgtgatc
     3721 ctggacaagg gagtggccat caagggcgac ctgatcatct ctatcgccgg cgtggacctg
     3781 gtgtacctgg atctgagagt gctgatctcc agcgtggaaa ccctggtgca ggccaaagag
     3841 ggcaaccaca agcccatcac cagcgagcag ttcgacaagc agaaagagga gctgatggac
     3901 gccaccggcc agcccagcaa gtggacaaat cctctgggca gcggcgctcc cgggtcaggt
     3961 gccacgaatt tttcgttgtt gaagcaagct ggggatgttg aagagaaccc agggcctgtg
     4021 cagcccgtgt cccaggccaa cggcagaatc cacctggatc ccgatcaggc cgaacaggga
     4081 ctggcccagc tcgtgatgac cgtgatcgag ctgctgcggc agatcgtgga acggcacgcc
     4141 atgagaagag tggaaggcgg caccctgacc gacgagcaga tcgagaatct gggaatcgct
     4201 ctgatgaacc tggaggagaa gatggacgag ctgaaagagg tgttcggact ggacgctgag
     4261 gatctgaaca tcgacctggg ccctctgggc agcctgctgg gtgccccggg atctggcgca
     4321 acaaatttta gtcttttaaa gcaggcagga gacgtcgagg aaaaccctgg acccgtggcc
     4381 gtggaacaca acatgcagag cagcaccatc gtggacgtgc tggaaaagat cctggacaag
     4441 ggcgtcgtga tcgccgggga catcacagtg ggaatcgccg acgtggaact gctgaccatc
     4501 aagatccggc tgatcgtggc cagcgtggac aaggccaaag aaatcggcat ggattggtgg
     4561 gagaacgacc cctacctgag cagcaagggc gccaacaaca aggctctgga agaggaaaac
     4621 aagatgctgc acgagcggct gaaaacactg gaagagaaga tcgagacaaa gcgcggggca
     4681 cctggctcgg gagcgaccaa cttctcatta ctcaaacaag ccggagacgt tgaggagaat
     4741 ccaggccctg tgagcaccgg ccccagcttc agcaccaagg acaacaccct ggaatacttc
     4801 gtgaaggcca gcaacaagca cggctttagc ctcgacatca gcctgaacgt gaatggggcc
     4861 gtgattagcg gcaccatgat cagcgccaaa gagtacttcg actacctgag cgagacattc
     4921 gaagagggca gcgaagtggc ccaggccctg tctgagcagt ttagcctggc tagcgaggcc
     4981 tccgagtcta atggcgaagc cgaggcccac ttcatccacc tgaagaacac caagatctac
     5041 tgcggcgaca gcaagagcac ccccagcaag gtcaagatct tctggcgcgg caagatcgcc
     5101 gaggtggacg gattcttcct gggaaaaatc agcgacgcca agtccaccag caagaagtcc
     5161 agcggcgctc ccgggtcagg tgccacgaat ttttcgttgt tgaagcaagc tggggatgtt
     5221 gaagagaacc cagggcctgt ggtgtccaag ggcgaggaac tgttcaccgg cgtggtgccc
     5281 atcctggtgg aactggatgg cgacgtgaac ggccacaagt tcagcgtgtc cggcgagggc
     5341 gaaggcgacg ccacatacgg aaagctgacc ctgaagttca tctgcaccac cggcaagctg
     5401 cccgtgcctt ggcctaccct cgtgaccaca ctgacctacg gcgtgcagtg cttcgccaga
     5461 taccccgacc acatgaagca gcacgatttc ttcaagagcg ccatgcccga gggctacgtg
     5521 caggaacgga ccatcttctt caaggacgac ggcaactaca agacaagagc cgaagtgaag
     5581 ttcgagggcg acaccctcgt gaaccggatc gagctgaagg gcatcgactt caaagaggat
     5641 ggcaacatcc tgggccacaa gctggagtac aactacaaca gccacaaggt gtacatcacc
     5701 gccgacaagc agaaaaacgg catcaaagtg aacttcaaga cccggcacaa catcgaggac
     5761 ggcagcgtgc agctggccga ccactaccag cagaacaccc ccatcggaga tggccccgtg
     5821 ctgctgcccg acaaccacta cctgagcaca caaagcgccc tgagcaagga ccccaacgag
     5881 aagcgggacc acatggtgct gctggaattt gtgaccgccg ctggcatcac cctgggcatg
     5941 gacgagctgt acaagtgaac tagtagacca cctcccctgc gagctaagct ggacagccaa
     6001 tgacgggtaa gagagtgaca tttttcacta acctaagaca ggagggccgt cagagctact
     6061 gcctaatcca aagacgggta aaagtgataa aaatgtatca ctccaaccta agacaggcgc
     6121 agcttccgag ggatttgaga tccagacatg ataagataca ttgatgagtt tggacaaacc
     6181 aaaactagaa tgcagtgaaa aaaatgcctt atttgtgaaa tttgtgatgc tattgcctta
     6241 tttgtaacca ttataagctg caataaacaa gtttgatatc tataacaaga aaatatatat
     6301 ataataagtt atcacgtaag tagaacatga aataacaata taattatcgt atgagttaaa
     6361 tcttaaaagt cacgtaaaag ataatcatgc gtcattttga ctcacgcggt cgttatagtt
     6421 caaaatcagt gacacttacc gcattgacaa gcacgcctca cgggagctcc aagcggcgac
     6481 tgagatgtcc taaatgcaca gcgacggatt cgcgctattt agaaagagag agcaatattt
     6541 caagaatgca tgcgtcaatt ttacgcagac tatctttcta gggttaagaa ttcactggcc
     6601 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     6661 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     6721 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     6781 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     6841 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     6901 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     6961 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     7021 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     7081 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     7141 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     7201 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     7261 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     7321 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     7381 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     7441 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     7501 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     7561 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     7621 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     7681 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     7741 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     7801 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     7861 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     7921 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     7981 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     8041 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     8101 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     8161 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     8221 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     8281 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     8341 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     8401 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     8461 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     8521 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     8581 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     8641 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     8701 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     8761 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     8821 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     8881 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     8941 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     9001 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     9061 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     9121 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     9181 taacaatttc acacaggaaa cagctatgac catgattacg ccaaggtcga cttaacccta
     9241 gaaagataat catattgtga cgtacgttaa agataatcat gcgtaaaatt gacgcatgtg
     9301 ttttatcggt ctgtatatcg aggtttattt attaatttga atagatatta agttttatta
     9361 tatttacact tacatactaa taataaattc aacaaacaat ttatttatgt ttatttattt
     9421 attaaaaaaa aacaaaaact caaaatttct tctataaagt aacaaaactt ttagcagtga
     9481 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     9541 tgcaataaac aagttaacta aacgggccct ctagatcagg caccgggctt gcgggtcatg
     9601 caccaggtgc gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc
     9661 tcgtagaagg ggaggttgcg gggcgcggag gtctccagga aggcgggcac cccggcgcgc
     9721 tcggccgcct ccactccggg gagcacgacg gcgctgccca gacccttgcc ctggtggtcg
     9781 ggcgagacgc cgacggtggc caggaaccac gcgggctcct tgggccggtg cggcgccagg
     9841 aggccttcca tctgttgctg cgcggccagc cgggaaccgc tcaactcggc catgcgcggg
     9901 ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg gcgtggtcca gaccgccacc
     9961 gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag
    10021 agttcttgca gctcggtgac ccgctcgatg tggcggtccg gatcgacggt gtggcgcgtg
    10081 gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg ccctggggac gtcgtcgcgg
    10141 gtggcgaggc gcaccgtggg cttgtactcg gtcatctcga gcctagggcc gggattctcc
    10201 tccacgtcac cgcatgttag aagacttcct ctgccctctc cgctgccccc ggggagcatg
    10261 tcaaggtcaa aatcgtcaag agcgtcagca ggcagcatat caaggtcaaa gtcgtcaagg
    10321 gcatcggctg ggagcatgtc taagtcaaaa tcgtcaaggg cgtcggtcgg cccgccgctt
    10381 tcgcacttta gctgtttctc caggccacat atgattagtt ccaggccgaa aaggaaggca
    10441 ggttcggctc cctgccggtc gaacagctca attgcttgtt tcagaagtgg gggcatagaa
    10501 tcggtggtag gtgtctctct ttcctctttt gctacttgat gctcctgttc ctccaatacg
    10561 cagcccagtg taaagtggcc cacggcggac agagcgtaca gtgcgttctc cagggagaag
    10621 ccttgctgac acaggaacgc gagctgattt tccagggttt cgtactgttt ctctgttggg
    10681 cgggtgccga gatgcacttt agccccgtcg cgatgtgaga ggagagcaca gcggtatgac
    10741 ttggcgttgt tccgcagaaa gtcttgccat gactcgcctt ccagggggca ggagtgggta
    10801 tgatgcctgt ccagcatctc gattggcagg gcatcgagca gggcccgctt gttcttcacg
    10861 tgccagtaca gggtaggctg ctcaactccc agcttttgag cgagtttcct tgtcgtcagg
    10921 ccttcgatac cgactccatt gagtaattcc agagcagagt ttatgacttt gctcttgtcc
    10981 agtctagaca tcttatcgtc atcgtctttg taatccatgg tggcggatcc cgcgtcacga
    11041 cacctgtgtt ctggcggcaa acccgttgcg aaaaagaac
//
