LOCUS       sequence-232860-       12255 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For targeted integration and inducible expression of human NODAL 
            (with an N-terminal MYC tag on the mature peptide) using 
            doxycycline.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12255)
  AUTHORS   Findlay SD, Bilyk O, Lypka K, Waskiewicz AJ, Postovit L
  TITLE     Genetically regulated human NODAL splice variants are differentially
            post-transcriptionally processed and functionally distinct
  JOURNAL   (2018) bioRxiv 276170
REFERENCE   2  (bases 1 to 12255)
  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: "(2018) 
            bioRxiv 276170"
COMMENT     Sequence Label: AAVS1-TRE3G-NODAL-matMYC
FEATURES             Location/Qualifiers
     source          1..12255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(3..21)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(42..58)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    66..82
                     /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)."
     promoter        complement(join(90..96,97..114,115..120))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    135..156
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(273..290)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(444..1032)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(524..543)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(1156..1224,1225..2016)
                     /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(1374..1393)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(2377..2396)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2991..3010
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(3413..3715)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(3515..3534)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     3838..3860
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3852..3869
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3853..3869
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3876..3895
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3876..3894
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3929..4765
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     protein_bind    4839..4860
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4875..4880,4881..4898,4899..4905)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4913..4929
                     /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     4937..4953
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5296..5315
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    5314..5369
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     5415..5434
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(5525..6271)
                     /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"
     primer_bind     complement(6309..6328)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          6336..7344
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(6390..6412)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(7345..7620)
                     /label=chicken beta-actin promoter
     enhancer        7623..7926
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8010..8388
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    8018..8036
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8054..8072
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8090..8108
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8126..8144
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8162..8180
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8198..8216
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8234..8252
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     8297..8321
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             9114..9143
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     intron          9637..9702
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             9832..9852
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    10277..10398
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10314..10333)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10368..10387
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    10436..10660
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     10649..10666
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(10698..11297)
                     /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(10781..10801)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     11278..11297
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(11307..11360)
                     /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"
     misc_feature    11384..11409
                     /label=SA
                     /note="splice acceptor site"
     misc_feature    11416..12219
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
ORIGIN
        1 tccctttagt gagggttaat tctgagcttg gcgtaatcat ggtcatagct gtttcctgtg
       61 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
      121 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
      181 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      241 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
      301 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
      361 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      421 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      481 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      541 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      601 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
      661 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
      721 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
      781 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
      841 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
      901 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
      961 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1021 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1081 aactcacgtt aagggatttt ggtcatgaga caataaccct gataaatgct tcaataatat
     1141 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     1201 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1261 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     1321 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     1381 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     1441 tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg
     1501 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     1561 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     1621 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     1681 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     1741 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     1801 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     1861 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     1921 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     1981 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     2041 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     2101 tttgataatc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     2161 gttccgcgca catttccccg aaaagtgcca cctgatgcgg tgtgaaatac cgcacagatg
     2221 cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta ataattcaga agaactcgtc
     2281 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
     2341 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
     2401 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
     2461 attttccacc atgatattcg gcaagcaggc atcgccatgc atgggtcacg acgagatcct
     2521 cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat
     2581 gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct
     2641 cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc
     2701 gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga
     2761 gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt
     2821 cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt
     2881 cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct
     2941 gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat
     3001 agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa
     3061 tcatgcgaaa cgatcctcat cctgtctctt gatcagagct tgatcccctg cgccatcaga
     3121 tccttggcgg caagaaagcc atccagttta ctttgcaggg cttcccaacc ttaccagagg
     3181 gcgccccagc tggcaattcc ggttcgcttg ctgtccataa aaccgcccag tctagctatc
     3241 gccatgtaag cccactgcaa gctacctgct ttctctttgc gcttgcgttt tcccttgtcc
     3301 agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct
     3361 acgtgaaaag gatctaggtg aagatccttt ttgataatct catgcctgac atttatattc
     3421 cccagaacat caggttaatg gcgtttttga tgtcattttc gcggtggctg agatcagcca
     3481 cttcttcccc gataacggag accggcacac tggccatatc ggtggtcatc atgcgccagc
     3541 tttcatcccc gatatgcacc accgggtaaa gttcacggga gactttatct gacagcagac
     3601 gtgcactggc cagggggatc accatccgtc gccccggcgt gtcaataata tcactctgta
     3661 catccacaaa cagacgataa cggctctctc ttttataggt gtaaacctta aactgccgta
     3721 cgtataggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     3781 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     3841 agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat
     3901 tgaatttagc ggccgcgaat tcgcccttag agcagagcca ggaacccctg tagggaaggg
     3961 gcaggagagc caggggcatg agatggtgga cgaggaaggg ggacagggaa gcctgagcgc
     4021 ctctcctggg cttgccaagg actcaaaccc agaagcccag agcagggcct tagggaagcg
     4081 ggaccctgct ctgggcggag gaatatgtcc cagatagcac tggggactct ttaaggaaag
     4141 aaggatggag aaagagaaag ggagtagagg cggccacgac ctggtgaaca cctaggacgc
     4201 accattctca caaagggagt tttccacacg gacacccccc tcctcaccac agccctgcca
     4261 ggacggggct ggctactggc cttatctcac aggtaaaact gacgcacgga ggaacaatat
     4321 aaattgggga ctagaaaggt gaagagccaa agttagaact caggaccaac ttattctgat
     4381 tttgtttttc caaactgctt ctcctcttgg gaagtgtaag gaagctgcag caccaggatc
     4441 agtgaaacgc accagacagc cgcgtcagag cagctcaggt tctgggagag ggtagcgcag
     4501 ggtggccact gagaaccggg caggtcacgc atccccccct tccctcccac cccctgccaa
     4561 gctctccctc ccaggatcct ctctggctcc atcgtaagca aaccttagag gttctggcaa
     4621 ggagagagat ggctccagga aatgggggtg tgtcaccaga taaggaatct gcctaacagg
     4681 aggtgggggt tagacccaat atcaggagac taggaaggag gaggcctaag gatggggctt
     4741 ttctgtcacc aatcctgtcc ctagtaaagc ttagtactgt cgatccgctg gcacgacagg
     4801 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     4861 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     4921 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct tgggctgcag
     4981 gtcgagggat ctccataaga gaagagggac agctatgact gggagtagtc aggagaggag
     5041 gaaaaatctg gctagtaaaa catgtaagga aaattttagg gatgttaaag aaaaaaataa
     5101 cacaaaacaa aatataaaaa aaatctaacc tcaagtcaag gcttttctat ggaataagga
     5161 atggacagca gggggctgtt tcatatactg atgacctctt tatagccaac ctttgttcat
     5221 ggcagccagc atatgggcat atgttgccaa actctaaacc aaatactcat tctgatgttt
     5281 taaatgattt gccctcccat atgtccttcc gagtgagaga cacaaaaaat tccaacacac
     5341 tattgcaatg aaaataaatt tcctttatta gccagaagtc agatgctcaa ggggcttcat
     5401 gatgtcccca taatttttgg cagagggaaa aagatctcag tggtatttgt gagccagggc
     5461 attggccaca ccagccacca ccttctgata ggcagcctgc acctgaggag tgaattcacg
     5521 cgccttaccc ggggagcatg tcaaggtcaa aatcgtcaag agcgtcagca ggcagcatat
     5581 caaggtcaaa gtcgtcaagg gcatcggctg ggagcatgtc taagtcaaaa tcgtcaaggg
     5641 cgtcggtcgg cccgccgctt tcgcacttta gctgtttctc caggccacat atgattagtt
     5701 ccaggccgaa aaggaaggca ggttcggctc cctgccggtc gaacagctca attgcttgtt
     5761 tcagaagtgg gggcatagaa tcggtggtag gtgtctctct ttcctctttt gctacttgat
     5821 gctcctgttc ctccaatacg cagcccagtg taaagtggcc cacggcggac agagcgtaca
     5881 gtgcgttctc cagggagaag ccttgctgac acaggaacgc gagctgattt tccagggttt
     5941 cgtactgttt ctctgttggg cgggtgccga gatgcacttt agccccgtcg cgatgtgaga
     6001 ggagagcaca gcggtatgac ttggcgttgt tccgcagaaa gtcttgccat gactcgcctt
     6061 ccagggggca ggagtgggta tgatgcctgt ccagcatctc gattggcagg gcatcgagca
     6121 gggcccgctt gttcttcacg tgccagtaca gggtaggctg ctcaactccc agcttttgag
     6181 cgagtttcct tgtcgtcagg ccttcgatac cgactccatt gagtaattcc agagcagagt
     6241 ttatgacttt gctcttgtcc agtctagaca tggtgctcga cgaattcttt gccaaaatga
     6301 tgagacagca caataaccag cacgttgccc aggagctgta ggaaaaagaa gaaggcatga
     6361 acatggttag cagaggctct agagccgccg gtcacacgcc agaagccgaa ccccgccctg
     6421 ccccgtcccc cccgaaggca gccgtccccc tgcggcagcc ccgaggctgg agatggagaa
     6481 ggggacggcg gcgcggcgac gcacgaaggc cctccccgcc catttccttc ctgccggcgc
     6541 cgcaccgctt cgcccgcgcc cgctagaggg ggtgcggcgg cgcctcccag atttcggctc
     6601 cgccagattt gggacaaagg aagtccctgc gccctctcgc acgattacca taaaaggcaa
     6661 tggctgcggc tcgccgcgcc tcgacagccg ccggcgctcc ggggccgccg cgcccctccc
     6721 ccgagccctc cccggcccga ggcggccccg ccccgcccgg cacccccacc tgccgccacc
     6781 ccccgcccgg cacggcgagc cccgcgccac gccccgcacg gagccccgca cccgaagccg
     6841 ggccgtgctc agcaactcgg ggaggggggt gcaggggggg gttacagccc gaccgccgcg
     6901 cccacacccc ctgctcaccc ccccacgcac acaccccgca cgcagccttt gttcccctcg
     6961 cagccccccc gcaccgcggg gcaccgcccc cggccgcgct cccctcgcgc acacgcggag
     7021 cgcacaaagc cccgcgccgc gcccgcagcg ctcacagccg ccgggcagcg cgggccgcac
     7081 gcggcgctcc ccacgcacac acacacgcac gcaccccccg agccgctccc ccccgcacaa
     7141 agggccctcc cggagccctt taaggctttc acgcagccac agaaaagaaa cgagccgtca
     7201 ttaaaccaag cgctaattac agcccggagg agaagggccg tcccgcccgc tcacctgtgg
     7261 gagtaacgcg gtcagtcaga gccggggcgg gcggcgcgag gcggcgcgga gcggggcacg
     7321 gggcgaaggc aacgcagcga ctcccgcccg ccgcgcgctt cgctttttat agggccgccg
     7381 ccgccgccgc ctcgccataa aaggaaactt tcggagcgcg ccgctctgat tggctgccgc
     7441 cgcacctctc cgcctcgccc cgccccgccc ctcgccccgc cccgccccgc ctggcgcgcg
     7501 cccccccccc ccccgccccc atcgctgcac aaaataatta aaaaataaat aaatacaaaa
     7561 ttgggggtgg ggaggggggg gagatgggga gagtgaagca gaacgtgggg ctcacctcga
     7621 cccatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag
     7681 gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc
     7741 gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatagtcc
     7801 acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt
     7861 gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat
     7921 gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga ttactattaa
     7981 taactagttt tcgtcttcaa gaattcctcg agtttactcc ctatcagtga tagagaacgt
     8041 atgaagagtt tactccctat cagtgataga gaacgtatgc agactttact ccctatcagt
     8101 gatagagaac gtataaggag tttactccct atcagtgata gagaacgtat gaccagttta
     8161 ctccctatca gtgatagaga acgtatctac agtttactcc ctatcagtga tagagaacgt
     8221 atatccagtt tactccctat cagtgataga gaacgtataa gctttaggcg tgtacggtgg
     8281 gcgcctataa aagcagagct cgtttagtga accgtcagat cgcctggagc aattccacaa
     8341 cacttttgtc ttataccaac tttccgtacc acttcctacc ctcgtaaagt cgacccacca
     8401 tgcacgccca ctgcctgccc ttccttctgc acgcctggtg ggccctactc caggcgggtg
     8461 ctgcgacggt ggccactgcg ctcctgcgta cgcgggggca gccctcgtcg ccatcccctc
     8521 tggcgtacat gctgagcctc taccgcgacc cgctgccgag ggcagacatc atccgcagcc
     8581 tacaggcaga agatgtggca gtggatgggc agaactggac gtttgctttt gacttctcct
     8641 tcctgagcca acaagaggat ctggcatggg ctgagctccg gctgcagctg tccagccctg
     8701 tggacctccc cactgagggc tcacttgcca ttgagatttt ccaccagcca aagcccgaca
     8761 cagagcaggc ttcagacagc tgcttagagc ggtttcagat ggacctattc actgtcactt
     8821 tgtcccaggt caccttttcc ttgggcagca tggttttgga ggtgaccagg cctctctcca
     8881 agtggctgaa gcaccctggg gccctggaga agcagatgtc cagggtagct ggagagtgct
     8941 ggccgcggcc ccccacaccg cctgccacca atgtgctcct tatgctctac tccaacctct
     9001 cgcaggagca gaggcagctg ggtgggtcca ccttgctgtg ggaagccgag agctcctggc
     9061 gggcccagga gggacagctg tcctgggagt ggggcaagag gcaccggcga catgagcaga
     9121 aactcatctc agaagaggat ctgcatcact tgccagacag aagtcaactg tgtcggaagg
     9181 tcaagttcca ggtggacttc aacctgatcg gatggggctc ctggatcatc taccccaagc
     9241 agtacaacgc ctatcgctgt gagggcgagt gtcctaatcc tgttggggag gagtttcatc
     9301 cgaccaacca tgcatacatc cagagtctgc tgaaacgtta ccagccccac cgagtccctt
     9361 ccacttgttg tgccccagtg aagaccaagc cgctgagcat gctgtatgtg gataatggca
     9421 gagtgctcct agatcaccat aaagacatga tcgtggaaga atgtgggtgc ctctgaacgc
     9481 gtcatatggc tagcctgcag ggatccaatg taactgtatt cagcgatgac gaaattctta
     9541 gctattgtaa tactctagag gatctttgtg aaggaacctt acttctgtgg tgtgacataa
     9601 ttggacaaac tacctacaga gatttaaagc tctaaggtaa atataaaatt tttaagtgta
     9661 taatgtgtta aactactgat tctaattgtt tgtgtatttt agattccaac ctatggaact
     9721 gatgaatggg agcagtggtg gaatgccttt aatgaggaaa acctgttttg ctcagaagaa
     9781 atgccatcta gtgatgatga ggctactgct gactctcaac attctactcc tccaaaaaag
     9841 aagagaaagg tagaagaccc caaggacttt ccttcagaat tgctaagttt tttgagtcat
     9901 gctgtgttta gtaatagaac tcttgcttgc tttgctattt acaccacaaa ggaaaaagct
     9961 gcactgctat acaagaaaat tatggaaaaa tattctgtaa cctttataag taggcataac
    10021 agttataatc ataacatact gttttttctt actccacaca ggcatagagt gtctgctatt
    10081 aataactatg ctcaaaaatt gtgtaccttt agctttttaa tttgtaaagg ggttaataag
    10141 gaatatttga tgtatagtgc cttgactaga gatcataatc agccatacca catttgtaga
    10201 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
    10261 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
    10321 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
    10381 actcatcaat gtatcttatc atgtctgcgg cctagtcaat aatcaatgtc gagacccata
    10441 gagcccaccg catccccagc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc
    10501 tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc
    10561 tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg
    10621 gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaggctga tcagcgggtt
    10681 taaacgggcc ctctagatca ggcaccgggc ttgcgggtca tgcaccaggt gcgcggtcct
    10741 tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc gctcgtagaa ggggaggttg
    10801 cggggcgcgg aggtctccag gaaggcgggc accccggcgc gctcggccgc ctccactccg
    10861 gggagcacga cggcgctgcc cagacccttg ccctggtggt cgggcgagac gccgacggtg
    10921 gccaggaacc acgcgggctc cttgggccgg tgcggcgcca ggaggccttc catctgttgc
    10981 tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg ggccgatctc ggcgaacacc
    11041 gcccccgctt cgacgctctc cggcgtggtc cagaccgcca ccgcggcgcc gtcgtccgcg
    11101 acccacacct tgccgatgtc gagcccgacg cgcgtgagga agagttcttg cagctcggtg
    11161 acccgctcga tgtggcggtc cggatcgacg gtgtggcgcg tggcggggta gtcggcgaac
    11221 gcggcggcga gggtgcgtac ggccctgggg acgtcgtcgc gggtggcgag gcgcaccgtg
    11281 ggcttgtact cggtcatctc gagcctaggg ccgggattct cctccacgtc accgcatgtt
    11341 agaagacttc ctctgccctc tccgctgcca gatctctcga ggccctgtgg gaggaagaga
    11401 agaggtcaga agcttgcccc actgtggggt ggaggggaca gataaaagta cccagaacca
    11461 gagccacatt aaccggccct gggaatataa ggtggtccca gctcggggac acaggatccc
    11521 tggaggcagc aaacatgctg tcctgaagtg gacatagggg cccgggttgg aggaagaaga
    11581 ctagctgagc tctcggaccc ctggaagatg ccatgacagg gggctggaag agctagcaca
    11641 gactagagag gtaagggggg taggggagct gcccaaatga aaggagtgag aggtgacccg
    11701 aatccacagg agaacggggt gtccaggcaa agaaagcaag aggatggaga ggtggctaaa
    11761 gccagggaga cggggtactt tggggttgtc cagaaaaacg gtgatgatgc aggcctacaa
    11821 gaaggggagg cgggacgcaa gggagacatc cgtcggagaa ggccatccta agaaacgaga
    11881 gatggcacag gccccagaag gagaaggaaa agggaaccca gcgagtgaag acggcatggg
    11941 gttgggtgag ggaggagaga tgcccggaga ggacccagac acggggagga tccgctcaga
    12001 ggacatcacg tggtgcagcg ccgagaagga agtgctccgg aaagagcatc cttgggcagc
    12061 aacacagcag agagcaaggg gaagagggag tggaggaaga cggaacctga aggaggcggc
    12121 agggaaggat ctgggccagc cgtagaggtg acccaggcca caagctgcag acagaaagcg
    12181 gcacaggccc aggggagaga atgctggtca gagaaagcaa agggcgaatt cgtttaaacc
    12241 tgcaggacta gctag
//
