LOCUS       sequence-426923-        9236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For integration of NGN2 WT T2A mEGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9236)
  AUTHORS   Naderi J, Magalhaes AP, Kibar G, Stik G, Zhang Y, Mackowiak SD, 
            Wieler HM, Rossi F, Buschow R, Christou-Kent M, Alcoverro-Bertran M,
            Graf T, Vingron M, Hnisz D
  TITLE     An activity-specificity trade-off encoded in human transcription 
            factors.
  JOURNAL   Nat Cell Biol. 2024 Aug;26(8):1309-1321. doi: 
            10.1038/s41556-024-01411-0. Epub 2024 Jul 5.
  PUBMED    38969762
REFERENCE   2  (bases 1 to 9236)
  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: 
            "10.1038/s41556-024-01411-0"; journalName: "Nat Cell Biol"; date: 
            "2024-08"; volume: "26"; issue: "8"; pages: "1309-1321"
COMMENT     Sequence Label: pPB-TetON-mEGFP-NGN2_WT
FEATURES             Location/Qualifiers
     source          1..9236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   147..209
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        658..1036
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    666..684
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    702..720
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    738..756
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    774..792
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    810..828
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    846..864
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    882..900
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     945..969
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1049..1058
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1058..1123
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1937..1948
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1940..1993
                     /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             join(2000..2002,2003..2713)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2000..2002,2003..2713)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2042..2063)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2303..2322)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2650..2671
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2800..2819)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2844..2892
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        3123..3334
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3347..3615
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3649..4392
                     /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             4393..4446
                     /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             4447..5046
                     /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(4447..4466)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4943..4963
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5079..5200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5116..5135)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5170..5189
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   5963..5997
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6488..6504)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6488..6504)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6501..6523)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6512..6528
                     /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(6536..6542,6543..6560,6561..6566))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6581..6602
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6719..6736)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6890..7478)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6970..6989)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7649..8440,8441..8509))
                     /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     8272..8291
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8510..8614)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8640..9095)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8777..8798)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8989..9008
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(9222..9236,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(9236,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgag cgcgcctcgt tcattcacgt ttttgaaccc gtggaggacg
       61 ggcagactcg cggtgcaaat gtgttttaca gcgtgatgga gcagatgaag atgctcgaca
      121 cgctgcagaa cacgcagcta gattaaccct agaaagataa tcatattgtg acgtacgtta
      181 aagataatca tgtgtaaaat tgacgcatgt gttttatcgg tctgtatatc gaggtttatt
      241 tattaatttg aatagatatt aagttttatt atatttacac ttacatacta ataataaatt
      301 caacaaacaa tttatttatg tttatttatt tattaaaaaa aacaaaaact caaaatttct
      361 tctataaagt aacaaaactt ttatgaggga cagccccccc ccaaagcccc cagggatgta
      421 attacgtccc tcccccgcta gggggcagca gcgagccgcc cggggctccg ctccggtccg
      481 gcgctccccc cgcatccccg agccggcagc gtgcggggac agcccgggca cggggaaggt
      541 ggcacgggat cgctttcctc tgaacgcttc tcgctgctct ttgagcctgc agacacctgg
      601 ggggatacgg ggaaaaggcc tccacggcca ctagtctttc gtcttcaaga attcctcgag
      661 tttactccct atcagtgata gagaacgtat gaagagttta ctccctatca gtgatagaga
      721 acgtatgcag actttactcc ctatcagtga tagagaacgt ataaggagtt tactccctat
      781 cagtgataga gaacgtatga ccagtttact ccctatcagt gatagagaac gtatctacag
      841 tttactccct atcagtgata gagaacgtat atccagttta ctccctatca gtgatagaga
      901 acgtataagc tttaggcgtg tacggtgggc gcctataaaa gcagagctcg tttagtgaac
      961 cgtcagatcg cctggagcaa ttccacaaca cttttgtctt ataccaactt tccgtaccac
     1021 ttcctaccct cgtaaaggtc tagagctagc caccatggac tacaaagacc atgacggtga
     1081 ttataaagat catgacatcg attacaagga tgacgatgac aagatgttcg tgaagtctga
     1141 aactctcgaa ctcaaagaag aggaagatgt tctggtcttg ctgggtagcg ctagcccagc
     1201 actggccgca ctcactcctc tcagcagcag tgcggatgag gaagaagaag aagaacccgg
     1261 ggcttccggc ggagcaagga gacaaagagg cgcagaagcg ggtcaaggtg cccgcggtgg
     1321 ggttgcggca ggtgccgaag gttgtaggcc agcgcgtttg cttgggttgg tgcatgactg
     1381 taagcgcagg cccagccgcg cacgagcagt gtctagagga gcgaaaacag cggaaacagt
     1441 acaaaggata aagaaaacga ggcgacttaa agcaaataat agagaacgca atagaatgca
     1501 taatttgaat gctgcccttg atgcccttcg tgaagtcctg cctacctttc ctgaagatgc
     1561 taaacttacg aaaatagaaa cactccggtt tgcacataat tatatttggg cgctgactga
     1621 aactttgaga ttggccgacc attgtggtgg tggtggtggc ggactcccag gcgccctgtt
     1681 tagcgaagcg gtcctcttgt cacccggcgg ggcatccgca gctttgtcct cttccggcga
     1741 tagtccatcc cctgcttcta cttggtcatg tacgaattct ccagccccat ctagcagtgt
     1801 cagctctaac tctacgagtc cttattcctg tacactgtcc ccagcgtccc ccgcgggctc
     1861 tgatatggat tactggcaac ctcctccccc tgacaagcac cgctatgcac ctcacctccc
     1921 catagccagg gattgtatcg agggcagagg aagtcttcta acatgcggtg acgtggagga
     1981 gaatcccggc cctagtaccg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     2041 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     2101 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     2161 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     2221 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     2281 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     2341 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     2401 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     2461 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     2521 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     2581 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     2641 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     2701 cgagctgtac aagaccggtt gaccggttag taatgagttt aaacggggga ggaagaccgg
     2761 ttgaccggtt agtaatgagt ttaaacgggg gaggctaact gaaacacgga aggagacaat
     2821 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg
     2881 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     2941 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     3001 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag
     3061 atctgcgaaa tcggatccag ctgaatctaa gtggatctgc gatcgctccg gtgcccgtca
     3121 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
     3181 aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
     3241 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
     3301 tctttttcgc aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc
     3361 tccttcacgc gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct
     3421 gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct
     3481 caggtcgaga ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct
     3541 ctccacgctt tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct
     3601 gcgccgttac agatctaagc tgtgaccggc gcctactcta gagccgccat gtctagactg
     3661 gacaagagca aagtcataaa ctctgctctg gaattactca atggagtcgg tatcgaaggc
     3721 ctgacgacaa ggaaactcgc tcaaaagctg ggagttgagc agcctaccct gtactggcac
     3781 gtgaagaaca agcgggccct gctcgatgcc ctgccaatcg agatgctgga caggcatcat
     3841 acccactcct gccccctgga aggcgagtca tggcaagact ttctgcggaa caacgccaag
     3901 tcataccgct gtgctctcct ctcacatcgc gacggggcta aagtgcatct cggcacccgc
     3961 ccaacagaga aacagtacga aaccctggaa aatcagctcg cgttcctgtg tcagcaaggc
     4021 ttctccctgg agaacgcact gtacgctctg tccgccgtgg gccactttac actgggctgc
     4081 gtattggagg aacaggagca tcaagtagca aaagaggaaa gagagacacc taccaccgat
     4141 tctatgcccc cacttctgaa acaagcaatt gagctgttcg accggcaggg agccgaacct
     4201 gccttccttt tcggcctgga actaatcata tgtggcctgg agaaacagct aaagtgcgaa
     4261 agcggcgggc cgaccgacgc ccttgacgat tttgacttag acatgctccc agccgatgcc
     4321 cttgacgact ttgaccttga tatgctgcct gctgacgctc ttgacgattt tgaccttgac
     4381 atgctccccg gggagggcag aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc
     4441 ggccctatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     4501 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     4561 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     4621 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     4681 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     4741 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     4801 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     4861 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     4921 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     4981 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     5041 gcctgaaatc aacctctgga ttacaaaatt tgtgggtaaa cttgtttatt gcagcttata
     5101 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5161 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg aattgactca
     5221 aatgatgtca attagtctat cagaagctca tctggtctcc cttccggggg acaagacatc
     5281 cctgtttaat atttaaacag cagtgttccc aaactgggtt cttatatccc ttgctctggt
     5341 caaccaggtt gcagggtttc ctgtcctcac aggaacgaag tccctaaaga aacagtggca
     5401 gccaggttta gccccggaat tgactggatt ccttttttag ggcccattgg tatggctttt
     5461 tccccgtatc cccccaggtg tctgcaggct caaagagcag cgagaagcgt tcagaggaaa
     5521 gcgatcccgt gccaccttcc ccgtgcccgg gctgtccccg cacgctgccg gctcggggat
     5581 gcggggggag cgccggaccg gagcggagcc ccgggcggct cgctgctgcc ccctagcggg
     5641 ggagggacgt aattacatcc ctgggggctt tggggggggg ctgtccctga tatctataac
     5701 aagaaaatat atatataata agttatcacg taagtagaac atgaaataac aatataatta
     5761 tcgtatgagt taaatcttaa aagtcacgta aaagataatc atgcgtcatt ttgactcacg
     5821 cggtcgttat agttcaaaat cagtgacact taccgcattg acaagcacgc ctcacgggag
     5881 ctccaagcgg cgactgagat gtcctaaatg cacagcgacg gattcgcgct atttagaaag
     5941 agagagcaat atttcaagaa tgcatgcgtc aattttacgc agactatctt tctagggtta
     6001 atctagctgc atcaggatca tatcgtcggg tcttttttcc ggctcagtca tcgcccaagc
     6061 tggcgctatc tgggcatcgg ggaggaagaa gcccgtgcct tttcccgcga ggttgaagcg
     6121 gcatggaaag agtttgccga ggatgactgc tgctgcattg acgttgagcg aaaacgcacg
     6181 tttaccatga tgattcggga aggtgtggcc atgcacgcct ttaacggtga actgttcgtt
     6241 caggccacct gggataccag ttcgtcgcgg cttttccgga cacagttccg gatggtcagc
     6301 ccgaagcgca tcagcaaccc gaacaatacc ggcgacagcc ggaactgccg tgccggtgtg
     6361 cagattaatg acagcggtgc ggcgctggga tattacgtca gcgaggacgg gtatcctggc
     6421 tggatgccgc agaaatggac atggataccc cgtgagttac ccggcgggcg cgcttggcgt
     6481 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6541 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6601 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6661 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6721 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6781 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6841 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6901 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6961 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     7021 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     7081 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7141 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7201 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7261 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7321 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7381 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7441 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7501 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7561 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7621 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7681 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7741 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7801 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7861 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7921 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7981 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     8041 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     8101 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     8161 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     8221 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     8281 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     8341 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     8401 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8461 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8521 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8581 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     8641 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     8701 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     8761 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     8821 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     8881 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     8941 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     9001 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     9061 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     9121 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     9181 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgtt
//
