LOCUS       sequence-476232-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible secreted embryonic alkaline phosphatase and mCherry; 
            constitutive BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  AUTHORS   Walton BL, Shattuck-Brandt R, Hamann CA, Tung VW, Colazo JM, Brand 
            DD, Hasty KA, Duvall CL, Brunger JM
  TITLE     A programmable arthritis-specific receptor for guided articular 
            cartilage regenerative medicine.
  JOURNAL   Osteoarthritis Cartilage. 2025 Feb;33(2):231-240. doi: 
            10.1016/j.joca.2024.12.002. Epub 2024 Dec 18.
  PUBMED    39706287
REFERENCE   2  (bases 1 to 13013)
  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.joca.2024.12.002"; journalName: "Osteoarthritis 
            Cartilage"; date: "2025-02"; volume: "33"; issue: "2"; pages: 
            "231-240"
COMMENT     Sequence Label: pHR TRE_SEAP_IRES_mCherry-PGK_BFP-hf
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    28..145
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        224..591
                     /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    232..250
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    268..286
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    304..322
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    340..358
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    376..394
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    412..430
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    448..466
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      630..639
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1470..1481)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    2207..2780
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2374..2391)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2601..2620
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2799..3509
                     /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(2944..2962)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3212..3231
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        3538..4037
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3573..3596)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3970..3989
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4058..4759
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    4855..5443
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4908..4928)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5326..5337)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5446..5462)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5447..5463)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5553..5786
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(5811..5829)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5812..5829)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6119..6138)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(6352..6371)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6471..6493
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6531..6549)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6617..6721
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6722..6790,6791..7582)
                     /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(6940..6959)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7753..8341
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8242..8261
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8495..8512
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(8582..8602)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8587..8916
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8767..8902
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          10050..10115
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             10245..10265
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    10690..10824
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10727..10746)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10781..10800
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             10992..11625
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    11672..11797
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12294..12527
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12712..12756
                     /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"
ORIGIN
        1 tcgccaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag
       61 tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag aattacaaaa
      121 acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg
      181 gatcgataag cttgatatcg aattcccgtg gaattatcac ctcgagttta ctccctatca
      241 gtgatagaga acgtatgaag agtttactcc ctatcagtga tagagaacgt atgcagactt
      301 tactccctat cagtgataga gaacgtataa ggagtttact ccctatcagt gatagagaac
      361 gtatgaccag tttactccct atcagtgata gagaacgtat ctacagttta ctccctatca
      421 gtgatagaga acgtatatcc agtttactcc ctatcagtga tagagaacgt ataagctttg
      481 cttatgtaaa ccagggcgcc tataaaagag tgctgatttt ttgagtaaac ttcaattcca
      541 caacactttt gtcttatacc aactttccgt accacttcct accctcgtaa agaattcccg
      601 tatacaccgg tgcaattctc acgcgtgccg ccaccatggt attgggtcca tgtatgcttt
      661 tgctcctgct gctgttgggc ttgcgccttc aactgtccct cggaataatt ccggtggaag
      721 aggagaatcc ggacttctgg aatagggagg cagccgaagc gcttggagca gcgaaaaaac
      781 ttcaaccggc ccagacggcc gccaagaatc tcataatttt tcttggcgat ggcatgggtg
      841 tttctacagt taccgcagct aggatcctga aggggcagaa aaaagacaag ttgggtcctg
      901 agatacccct cgcgatggat agattcccat atgttgcgct gagtaagaca tacaatgtcg
      961 acaaacatgt gcctgattca ggcgcgacag cgacggccta cctctgtgga gtaaaaggaa
     1021 acttccagac tattgggctt agtgcagcag ccagatttaa tcagtgtaat acgaccagag
     1081 gcaacgaggt aataagtgtc atgaatagag cgaagaaagc tggcaaatct gtcggtgtag
     1141 taactacaac cagggttcag cacgcttcac ccgctggtac ttacgcgcac acagttaata
     1201 gaaactggta tagtgatgct gacgttcccg cctctgccag gcaggaaggt tgccaggaca
     1261 tagccaccca gctgatttcc aacatggaca tagacgttat cttgggagga ggtaggaaat
     1321 acatgtttag aatggggacc ccagacccag aatacccgga cgattactct caggggggaa
     1381 caaggctgga cgggaagaat ctggttcaag agtggcttgc gaaaagacag ggtgcacggt
     1441 acgtttggaa taggactgag ctcatgcagg cgtccctcga tcccagtgtt acacatctca
     1501 tgggcctgtt tgagccgggc gacatgaaat atgaaattca cagagattca accctggatc
     1561 cgtctcttat ggaaatgaca gaagcagcgc tcagactcct gtcaagaaat cccaggggat
     1621 ttttcctttt tgttgagggg ggacgaattg accacggcca tcatgagtcc cgagcatatc
     1681 gggccctcac agagactatc atgttcgatg acgcaataga aagggcaggg cagctcactt
     1741 ccgaggagga cactctgagt ttggtcacag cggatcattc acatgtcttt tccttcggcg
     1801 gctaccccct tcgggggtct tctatttttg ggctggctcc cggaaaggcc agggatagga
     1861 aagcatacac ggtgctgctg tacggtaatg gccccggtta cgtcctgaaa gacggggccc
     1921 ggcctgatgt tacggaatct gagtccggat cccctgaata tagacagcag tccgcggtcc
     1981 ctctcgatga agaaactcac gcgggggagg atgtagcagt ttttgctcgc gggccccagg
     2041 cacatcttgt ccacggggtt caggagcaga cctttatcgc tcatgtgatg gcctttgctg
     2101 catgtcttga gccgtacacg gcgtgcgatt tggctccccc agcgggtact acagacgccg
     2161 cgcacccagg gagaagtcgc agcaaacgct tggattgagc ggccgccccc tctccctccc
     2221 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata
     2281 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     2341 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
     2401 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag
     2461 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     2521 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     2581 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     2641 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat
     2701 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     2761 tttgaaaaac acgatgataa atgagcgctg gcgggtccat ggtgagcaag ggcgaggagg
     2821 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
     2881 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     2941 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
     3001 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     3061 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     3121 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
     3181 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
     3241 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
     3301 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     3361 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
     3421 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
     3481 ccggcggcat ggacgagctg tacaagtagg gccgcctcga ggttaacgaa ttctaccggg
     3541 taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca
     3601 cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc
     3661 aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga
     3721 agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc
     3781 tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg
     3841 gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg
     3901 ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg
     3961 gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct
     4021 catctccggg cctttcgacc tgcagcccaa gcttaccatg agcgagctga ttaaggagaa
     4081 catgcacatg aagctgtaca tggagggcac cgtggacaac catcacttca agtgcacatc
     4141 cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg tggtcgaggg
     4201 cggccctctc cccttcgcct tcgacatcct ggctactagc ttcctctacg gcagcaagac
     4261 cttcatcaac cacacccagg gcatccccga cttcttcaag cagtccttcc ctgagggctt
     4321 cacatgggag agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac
     4381 cagcctccag gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcacatc
     4441 caacggccct gtgatgcaga agaaaacact cggctgggag gccttcaccg agacgctgta
     4501 ccccgctgac ggcggcctgg aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag
     4561 ccatctgatc gcaaacatca agaccacata tagatccaag aaacccgcta agaacctcaa
     4621 gatgcctggc gtctactatg tggactacag actggaaaga atcaaggagg ccaacaacga
     4681 gacctacgtc gagcagcacg aggtggcagt ggccagatac tgcgacctcc ctagcaaact
     4741 ggggcacaag cttaattaag gccgcaactc ccacctgcaa catgcgtgac tgactgaggc
     4801 cgcgactcta gagtcgacct gcaggcatgc aagcttgata tcaagcttat cgataatcaa
     4861 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4921 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4981 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     5041 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     5101 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     5161 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     5221 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     5281 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     5341 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     5401 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct
     5461 cgagggaatt aattcgagct cggtaccttt aagaccaatg acttacaagg cagctgtaga
     5521 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
     5581 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     5641 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     5701 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     5761 ttagtcagtg tggaaaatct ctagcagcat ctagaattaa ttccgtgtat tctatagtgt
     5821 cacctaaatc gtatgtgtat gatacataag gttatgtatt aattgtagcc gcgttctaac
     5881 gacaatatgt acaagcctaa ttgtgtagca tctggcttac tgaagcagac cctatcatct
     5941 ctctcgtaaa ctgccgtcag agtcggtttg gttggacgaa ccttctgagt ttctggtaac
     6001 gccgtcccgc acccggaaat ggtcagcgaa ccaatcagca gggtcatcgc tagccagatc
     6061 ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc
     6121 tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct
     6181 tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc
     6241 ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc
     6301 ttcctaatgc aggagtcgca taagggagag cgtcgaatgg tgcactctca gtacaatctg
     6361 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     6421 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     6481 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     6541 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     6601 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     6661 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     6721 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     6781 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     6841 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     6901 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     6961 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     7021 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     7081 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     7141 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     7201 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     7261 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     7321 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     7381 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     7441 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     7501 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     7561 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     7621 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     7681 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     7741 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     7801 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     7861 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     7921 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     7981 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     8041 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     8101 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     8161 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     8221 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     8281 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     8341 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     8401 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     8461 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     8521 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     8581 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     8641 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     8701 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     8761 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     8821 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     8881 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttggacacaa gacaggcttg
     8941 cgagatatgt ttgagaatac cactttatcc cgcgtcaggg agaggcagtg cgtaaaaaga
     9001 cgcggactca tgtgaaatac tggtttttag tgcgccagat ctctataatc tcgcgcaacc
     9061 tattttcccc tcgaacactt tttaagccgt agataaacag gctgggacac ttcacatgag
     9121 cgaaaaatac atcgtcacct gggacatgtt gcagatccat gcacgtaaac tcgcaagccg
     9181 actgatgcct tctgaacaat ggaaaggcat tattgccgta agccgtggcg gtctgtaccg
     9241 ggtgcgttac tggcgcgtga actgggtatt cgtcatgtcg ataccgtttg tatttccagc
     9301 tacgatcacg acaaccagcg cgagcttaaa gtgctgaaac gcgcagaagg cgatggcgaa
     9361 ggcttcatcg ttattgatga cctggtggat accggtggta ctgcggttgc gattcgtgaa
     9421 atgtatccaa aagcgcactt tgtcaccatc ttcgcaaaac cggctggtcg tccgctggtt
     9481 gatgactatg ttgttgatat cccgcaagat acctggattg aacagccgtg ggatatgggc
     9541 gtcgtattcg tcccgccaat ctccggtcgc taatcttttc aacgcctggc actgccgggc
     9601 gttgttcttt ttaacttcag gcgggttaca atagtttcca gtaagtattc tggaggctgc
     9661 atccatgaca caggcaaacc tgagcgaaac cctgttcaaa ccccgcttta aacatcctga
     9721 aacctcgacg ctagtccgcc gctttaatca cggcgcacaa ccgcctgtgc agtcggccct
     9781 tgatggtaaa accatccctc actggtatcg catgattaac cgtctgatgt ggatctggcg
     9841 cggcattgac ccacgcgaaa tcctcgacgt ccaggcacgt attgtgatga gcgatgccga
     9901 acgtaccgac gatgatttat acgatacggt gattggctac cgtggcggca actggattta
     9961 tgagtgggcc ccggatcttt gtgaaggaac cttacttctg tggtgtgaca taattggaca
    10021 aactacctac agagatttaa agctctaagg taaatataaa atttttaagt gtataatgtg
    10081 ttaaactact gattctaatt gtttgtgtat tttagattcc aacctatgga actgatgaat
    10141 gggagcagtg gtggaatgcc tttaatgagg aaaacctgtt ttgctcagaa gaaatgccat
    10201 ctagtgatga tgaggctact gctgactctc aacattctac tcctccaaaa aagaagagaa
    10261 aggtagaaga ccccaaggac tttccttcag aattgctaag ttttttgagt catgctgtgt
    10321 ttagtaatag aactcttgct tgctttgcta tttacaccac aaaggaaaaa gctgcactgc
    10381 tatacaagaa aattatggaa aaatattctg taacctttat aagtaggcat aacagttata
    10441 atcataacat actgtttttt cttactccac acaggcatag agtgtctgct attaataact
    10501 atgctcaaaa attgtgtacc tttagctttt taatttgtaa aggggttaat aaggaatatt
    10561 tgatgtatag tgccttgact agagatcata atcagccata ccacatttgt agaggtttta
    10621 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
    10681 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
    10741 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
    10801 aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa
    10861 cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc
    10921 tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca
    10981 aacttagtag ttggaagggc taattcactc ccaaagaaga caagatatcc ttgatctgtg
    11041 gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt
    11101 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt
    11161 agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat
    11221 ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca
    11281 cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa
    11341 gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg
    11401 agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag
    11461 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
    11521 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
    11581 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag
    11641 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga
    11701 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
    11761 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
    11821 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
    11881 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
    11941 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
    12001 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
    12061 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
    12121 caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga tatgagggac
    12181 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
    12241 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
    12301 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
    12361 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
    12421 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
    12481 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
    12541 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
    12601 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
    12661 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
    12721 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
    12781 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
    12841 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
    12901 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
    12961 ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg gta
//
