LOCUS       sequence-438466-       11934 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SEMPER plasmid for expression of gas vesicles using mammalian 
            acoustic reporter genes (mARGs).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11934)
  AUTHORS   Duan M, Dev I, Lu A, Ayrapetyan G, You MY, Shapiro MG
  TITLE     SEMPER: Stoichiometric expression of mRNA polycistrons by eukaryotic
            ribosomes for compact, ratio-tunable multi-gene expression.
  JOURNAL   Cell Syst. 2024 Jul 17;15(7):597-609.e4. doi: 
            10.1016/j.cels.2024.06.001. Epub 2024 Jul 5.
  PUBMED    38971149
REFERENCE   2  (bases 1 to 11934)
  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"; doi: 
            "10.1016/j.cels.2024.06.001"; journalName: "Cell Syst"; date: 
            "2024-07-17- 17"; volume: "15"; issue: "7"; pages: "597-609.e4"
COMMENT     Sequence Label: 
            pCMV-SEMPER-mARG_ACC-gvpA_ACC-gvpNJKFGW-EmGFP_IRES-mCherry
FEATURES             Location/Qualifiers
     source          1..11934
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     758..775
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(849..953)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1021..1039
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1066..1071,1072..1089,1090..1096)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1104..1120
                     /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     1109..1131
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     enhancer        1150..1453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1454..1657
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1608..1628
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1654..1678
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2995..3051
                     /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             3814..3870
                     /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             4354..4410
                     /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             5164..5220
                     /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             5596..5652
                     /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             6349..6405
                     /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(6712..6731)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    7160..7711
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7304..7321)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7531..7550
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7723..8433
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(7868..7886)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     8136..8155
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(8487..8506)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8488..8506)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    8901..9489
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8954..8974)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9372..9383)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    9509..9985
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(9708..9727)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      10121..10575
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10208..10227)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10417..10438
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             join(10862..10930,10931..11722)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAVLGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(11080..11099)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    11824..11857
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttattcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
      721 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      781 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      841 agcttgcaat tcgcgctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
      901 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
      961 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     1021 aataggcgta tcacgaggcc ctttcactca ttaggcaccc caggctttac actttatgct
     1081 tccggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcat
     1141 cgtgcaggtc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     1201 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     1261 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     1321 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     1381 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     1441 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     1501 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     1561 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     1621 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc
     1681 ctggagacgc catccacgct gttttgacct ccatagaaga ccgagctcgg atccagtacc
     1741 cttcaccatg gccgtggaaa agaccaacag cagcagctcc ctggccgaag tgatcgacag
     1801 aatcctggac aagggcatcg tgatcgacgc ctgggtgcgc gtgtccctcg tgggaattga
     1861 gctgctggcc atcgaggccc ggatcgtgat tgccagcgtg gaaacctacc tgaagtacgc
     1921 cgaggccgtg ggcctgacac agagtgctgc tgtgcctgct tgattaccat ggtgaccacc
     1981 accaaagtga accacaagcg ggccgtgctg agactgaggc ctggccagtt tgtcgtgacc
     2041 cccgccattg agcgggtggc cattagagcc ctgagatacc tgaagtccgg cttccccgtg
     2101 cacctgagag gacctgccgg aaccggcaag accacactgg ccatgcacct ggccaactgc
     2161 ctggacagac ccgtgatgct gctgttcggc gacgaccagt tcaagagcag cgacctgatc
     2221 ggcagcgaga gcggctacac ccacaagaag gtgctggaca actacatcca cagcgtcgtg
     2281 aagctggaag atgagttcaa gcagaactgg gtggacagca gactgaccct ggcctgccgg
     2341 gaaggcttca ccctggtgta cgacgagttc aaccggtcca gacccgaagt gaacaacgtg
     2401 ctgctgagcg ccctggaaga gaagatcctg agcctgcccc ccagcagcaa ccagccagag
     2461 tacctgagcg tgaaccccca gttcagagtg atcttcacca gcaaccccga ggaatatgcc
     2521 ggcgtgcaca gcacccagga cgccctgatg gaccggctcg tgaccatctc catgcccgag
     2581 cccgatgaga tcacccagac cgagatcctg atccagaaaa caaacatcga ccgcgagagc
     2641 gccaacttca tcgtgcggct cgtgaagtcc ttcagactgg ccacaggcgc cgagaaaacc
     2701 agcggcctga gaagctgcct gatgatcgcc aaagtgtgcg ccgacaacaa catccccgtg
     2761 accaccgaga gcctggactt ccccgatatc gccatcgaca tcctgttcaa ccgcagccac
     2821 ctgagcatga gcgagtccac caacatcttt ctggaactgc tggataagtt cagcgccgag
     2881 gaactggaaa tcctgaacaa cagagtgacc ggcgacaacg acttcctgat cgacaacagc
     2941 cagttcgtgt cccagcagct ggccggacag cccaacggag cgccaggttc cggggctact
     3001 aacttcagcc tccttaaaca ggctggcgat gtggaagaaa atcccggacc agtgctgccc
     3061 accagacccc agaccaacag cagccggacc atcaacacca gcacccaggg cagcaccctg
     3121 gccgacatcc tggaaagagt gctggacaag ggcatcgtga tcgccgggga catcagcatc
     3181 tctatcgcca gcaccgagct ggtgcacatc cggatcagac tgctgatctc cagcgtggac
     3241 aaggccaaag agatgggcat caactggtgg gagagcgacc cctacctgag caccaaggcc
     3301 cagagactgg tggaagagaa ccagcagctc cagcacagac tggaaagcct ggaagccaag
     3361 ctgaacagcc tgaccagcag cagcgtgaaa gaggaaatcc ccctggccgc cgacgtgaag
     3421 gacgacctgt atcagaccag cgccaagatc cccagccccg tggatacccc tatcgaggtg
     3481 ctggacttcc aggcccagtc tagcggcggc acaccccctt acgtgaacac ctctatggaa
     3541 atactggatt ttcaggctca gacctccgcc gagagcagca gccctgtggg ctctaccgtg
     3601 gaaatactgg acttccaggc acagacaagc gaggaatcca gctcccccgt ggtgtccaca
     3661 gtggaaatac tggatttcca ggcccagact tctgaagagt ccagctctcc agtgggaagc
     3721 actgtggaaa tactggactt tcaggcccag accagtgaag agatccccag ctctgtggac
     3781 cccgccatcg atgtgggtgc cccgggatct ggcgcaacaa attttagtct cctcaagcag
     3841 gcaggagatg tcgaggaaaa ccctggaccc gtggtgtgca cccctgccga gaacttcaac
     3901 aacagcctga caatcgccag caagcccaag aacgaggccg gactggctcc tctgctgctg
     3961 accgtgctgg aactcgtgcg gcagctgatg gaagcccaag tgatccggcg gatggaagag
     4021 gacctgctga gcgagcccga cctggaaaga gccgccgata gcctccagaa gctggaagaa
     4081 cagatcctgc acctgtgcga gatgttcgag gtggaccccg ccgacctgaa catcaacctg
     4141 ggcgagatcg gcaccctgct gcctagcagc ggcagctact atcctggcca gccaagcagc
     4201 agaccctctg tgctggaact gctggaccgg ctgctgaaca ccggcatcgt ggtggacggc
     4261 gagattgacc tgggaatcgc ccagatcgac ctgatccacg ccaagctgag actggtgctg
     4321 acctccaagc ccatcggggc acctggctca ggagcgacca acttctcatt gctcaaacaa
     4381 gccggagatg ttgaggagaa tccaggccct gtgagcatcc ccctgtacct gtacggcatc
     4441 ttccccaaca ccatccccga gacactggaa ctggaaggcc tggacaagca gcccgtgcac
     4501 agccaggtgg tggacgagtt ctgcttcctg tacagcgagg cccggcagga aaagtacctg
     4561 gccagcagac ggaacctgct gacccacgag aaggtgctgg aacagaccat gcacgccggc
     4621 ttcagagtgc tgctgcccct gagatttggc ctggtcgtga aggactggga gacaatcatg
     4681 agccagctga tcaaccccca caaggaccag ctgaaccagc tgttccagaa gctggccggc
     4741 aagcgggaag tgtccatcaa gattttctgg gacgccaagg ccgagcttca gaccatgatg
     4801 gaaagccacc aggacctgaa gcagcagcgg gacaacatgg aaggcaagaa actgagcatg
     4861 gaagaagtga tccagatcgg gcagctgatc gagatcaacc tgctggcccg gaagcaggcc
     4921 gtgatcgagg tgttcagcca ggaactgaac cccttcgccc aggaaatcgt ggtgtccgac
     4981 cccatgaccg aggaaatgat ctacaacgcc gccttcctga tcccctggga gagcgagagc
     5041 gagttcagcg agcgggtgga agtgatcgac cagaagttcg gcgaccggct gcggatcaga
     5101 tacaacaact tcaccgcccc ctacaccttt gcccagctgg actctggcgc tcccgggtca
     5161 ggtgccacta atttttcatt gttgaagcaa gctggggatg ttgaagagaa cccagggcct
     5221 gtgctgacca aactcctgct gctgcccatc atgggacccc tgaatggcgt cgtgtggatc
     5281 gccgagcaga tccaggaacg gaccaacacc gagttcgacg cccaggaaaa cctgcacaaa
     5341 cagctgctga gcctccagct gagcttcgac atcggcgaga tcggagagga agagttcgag
     5401 attcaggaag aggaaatcct gctgaagatc caggccctgg aagaagaggc ccggctggaa
     5461 ctggaagccg agcaggaaga agccagactg gaactggaag ctgaacagga agatttcgag
     5521 tacccaccac agttcaccgc cgaagtgaac aaggatcagc atctggtgct gctgcccggt
     5581 gccccgggat ctggcgctac taacttcagc ctccttaaac aggctggcga tgtggaagaa
     5641 aatcccggac cagtggaact ggaaaacctg tacacctacg cctttctgga aatccccagc
     5701 agccccctga tcctgcctca gggcgctgct aatcaggtgg tgctgatcaa cggcaccgag
     5761 ctggccgcca tcgtggaacc tggcatcttc ctggaatcct tccagaacaa cgacgagaag
     5821 atcatccaga tggccctgag ccacgacaga gtgatctgcg agctgttcca gcagatcacc
     5881 gtgctgcccc tgagattcgg cacctacttc accagcacca acaacctgct gaaccacctg
     5941 aagtcccacg agaaagagta ccagaacaag ctggaaaaga tcaacgggaa gaacgagttc
     6001 accctgaagc tgatcccccg gatgatcgag gaaatcgtgc cctctgaggg cggaggcaag
     6061 gactacttcc tggccaagaa gcagcgctac cagaatcaga acaacttctc tatcgcccag
     6121 gccgccgaga agcagaacct gatcgacctg atcaccaaag tgaaccagct gcccgtggtg
     6181 gtgcaggaac aggaagaaca gatccaaatc tacctgctgg tgtcctgcca ggataagacc
     6241 ctgctgctgg aacagttcct gacctggcag aaagcctgcc ccaagtggga tctgctgctg
     6301 ggcgactgcc tgccccccta ccactttatc ggagcgccag gttccggggc aacaaatttt
     6361 agtctcctca agcaggcagg agatgtcgag gaaaaccctg gacccgtggt gtccaagggc
     6421 gaggaactgt tcaccggcgt ggtgcccatc ctggtggaac tggatggcga cgtgaacggc
     6481 cacaagttca gcgtgtccgg cgagggcgaa ggcgacgcca catacggaaa gctgaccctg
     6541 aagttcatct gcaccaccgg caagctgccc gtgccttggc ctaccctcgt gaccacactg
     6601 acctacggcg tgcagtgctt cgccagatac cccgaccaca tgaagcagca cgatttcttc
     6661 aagagcgcca tgcccgaggg ctacgtgcag gaacggacca tcttcttcaa ggacgacggc
     6721 aactacaaga caagagccga agtgaagttc gagggcgaca ccctcgtgaa ccggatcgag
     6781 ctgaagggca tcgacttcaa agaggatggc aacatcctgg gccacaagct ggagtacaac
     6841 tacaacagcc acaaggtgta catcaccgcc gacaagcaga aaaacggcat caaagtgaac
     6901 ttcaagaccc ggcacaacat cgaggacggc agcgtgcagc tggccgacca ctaccagcag
     6961 aacaccccca tcggagatgg ccccgtgctg ctgcccgaca accactacct gagcacacaa
     7021 agcgccctga gcaaggaccc caacgagaag cgggaccaca tggtgctgct ggaatttgtg
     7081 accgccgctg gcatcaccct gggcatggac gagctgtaca agtgataata ctaaatcgca
     7141 ctgtcggcgt ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     7201 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     7261 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     7321 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     7381 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     7441 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     7501 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     7561 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     7621 tgctttacat gtgtttagtc gaggttaaaa aacgtctagg ccccccgaac cacggggacg
     7681 tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggtgag caagggcgag
     7741 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
     7801 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     7861 cagaccgcca agctgaaggt gaccaagggc ggccccctgc ccttcgcctg ggacatcctg
     7921 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
     7981 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     8041 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     8101 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     8161 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
     8221 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
     8281 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
     8341 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     8401 tccaccggcg gcatggatga gctgtacaag tgaaggatcc ctcgaggggc ccaagcttac
     8461 gcgtgcatgc gacgtcatag ctctctccct atagtgagtc gtattataag ctagcttggg
     8521 atctttgtga aggaacctta cttctgtggt gtgacataat tggacaaact acctacagag
     8581 atttaaagct ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa actagctgca
     8641 tatgcttgct gcttgagagt tttgcttact gagtatgatt tatgaaaata ttatacacag
     8701 gagctagtga ttataattgt ttgtgtattt tagattcaca gtcccaaggc tcatttcagg
     8761 cccctcagtc ctcacagtct gttcatgatc ataatcagcc ataccacatt tgtagaggtt
     8821 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     8881 attgttgttg ttcgtggctg aatcaacctc tggattacaa aatttgtgaa agattgactg
     8941 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     9001 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     9061 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     9121 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     9181 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     9241 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     9301 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     9361 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     9421 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     9481 cctccccgcc agagacaatt aacttcgcgg gtggcatccc tgtgacccct ccccagtgcc
     9541 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     9601 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     9661 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     9721 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     9781 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
     9841 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
     9901 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
     9961 aaccactgct cccttccctg tccttgaagt tctcagatcc tgcattaatg aatcggccaa
    10021 cgcgcgggga gaggcggttt gcgtattggc tggcgtaata gcgaagaggc ccgcaccgat
    10081 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
    10141 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
    10201 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
    10261 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
    10321 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
    10381 tttcgccctt tgacgttgga gtccacgttc ttaatagtgg actcttgttc caaactggaa
    10441 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
    10501 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
    10561 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
    10621 atttttctaa atacattcaa atatgtatcc gctcatgcca ggtcttggac tggtgagaac
    10681 ggcttgctcg gcagcttcga tgtgtgctgg agggagaata aaggtctaag atgtgcgata
    10741 gagggaagtc gcattgaatt atgtgctgtg tagggatcgc tggtatcaaa tatgtgtgcc
    10801 cacccctggc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
    10861 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
    10921 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
    10981 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
    11041 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
    11101 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
    11161 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
    11221 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
    11281 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
    11341 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
    11401 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
    11461 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
    11521 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
    11581 tcgcggtatc attgcagtac tggggccaga tggtaagccc tcccgtatcg tagttatcta
    11641 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
    11701 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
    11761 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
    11821 gccataactt cgtataatgt atgctatacg aagttatggc atgaccaaaa tcccttaacg
    11881 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttc
//
