LOCUS       sequence-403386-       13219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Improved Dox-inducible neurogenin-2; Piggybac Tet-ON plasmid for 
            differentiating hiPSCs into glutamatergic neurons via NGN2 
            expression. oNGN2 is a Phospho-mutant which avoids inhibition by cdk
            kinase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13219)
  TITLE     Characterization of iPSC lines for the iPSC Neurodegenerative 
            Disease Initiative (iNDI)
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 13219)
  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: 
            "Unpublished"
COMMENT     Sequence Label: iF59 PB-TO-oNgn2-puro
FEATURES             Location/Qualifiers
     source          1..13219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     351..370
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    369..424
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     470..489
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(580..1326)
                     /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(1358..1377)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          1385..2401
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(1439..1461)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(2402..2679)
                     /label=chicken beta-actin promoter
     enhancer        2681..3060
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    3061..3109
                     /note="synthetic polyadenylation signal"
     misc_feature    3123..3214
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3163..3182
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     protein_bind    complement(4426..4459)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4460..4671
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          4672..5610
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5567..5587
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     polyA_signal    6248..6472
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(6473..6506)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   7022..7056
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7547..7563)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7571..7587
                     /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(7595..7601,7602..7619,7620..7625))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7640..7661
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7949..8537)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8029..8048)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8708..9499,9500..9568))
                     /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     9331..9350
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9569..9673)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(9699..10154)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9836..9857)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10048..10067
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10281..10303
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10295..10312
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     10296..10312
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   10442..10504
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        11609..11986
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    11616..11634
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11652..11670
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11688..11706
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11724..11742
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11760..11778
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11796..11814
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11832..11850
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     11895..11919
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     polyA_signal    12860..12981
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12897..12916)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12951..12970
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13030..13046)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    13054..13070
                     /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(13078..13084,13085..13102,13103..13108))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13123..13144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
       61 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggacgg ccgcacatgt
      121 aaggaaaatt ttagggatgt taaagaaaaa aataacacaa aacaaaatat aaaaaaaatc
      181 taacctcaag tcaaggcttt tctatggaat aaggaatgga cagcaggggg ctgtttcata
      241 tactgatgac ctctttatag ccaacctttg ttcatggcag ccagcatatg ggcatatgtt
      301 gccaaactct aaaccaaata ctcattctga tgttttaaat gatttgccct cccatatgtc
      361 cttccgagtg agagacacaa aaaattccaa cacactattg caatgaaaat aaatttcctt
      421 tattagccag aagtcagatg ctcaaggggc ttcatgatgt ccccataatt tttggcagag
      481 ggaaaaagat ctcagtggta tttgtgagcc agggcattgg ccacaccagt caccaccttc
      541 tgataggcag cctgcacctg aggagtgaat tcacgcgttt tagcctggca gcatatcgag
      601 atcgaagtca tcgagagcgt cagcagggag catatccaga tcaaaatcat ccagggcatc
      661 ggcgggcagc atgtccaggt cgaagtcgtc cagggcgtcg gtgggtccgc cgctctcgca
      721 cttcagctgc ttctcgaggc cgcagatgat cagttccagg ccgaacagga aggcgggctc
      781 ggcgccctgt ctgtcgaaca gctcgatggc ctgcttcagc agggggggca tgctgtcggt
      841 ggtgggtgtc tctctttcct ctttggcgac ctggtgttcc tgttcttcca gcacgcagcc
      901 cagggtgaag tggcccacgg cgctcagggc gtacagggcg ttttccaggc tgaagccctg
      961 ctggcacagg aaggccagct ggttttccag tgtctcgtac tgcttctcgg tgggtctggt
     1021 gcccaggtgc actttggcgc cgtcccggtg gctcagcagg gcgcatctgt agctcttggc
     1081 gttgttccgc aggaagtcct gccagctctc gccttccaga gggcagctgt gggtgtggtg
     1141 ccggtccagc atctcgatgg gcagggcgtc cagcagggcc cgcttgttct tcacgtgcca
     1201 gtacagggtg ggctgttcca cgcccagttt ctgggccagc ttccgggtgg tcaggccctc
     1261 gatgcccacg ccgttcagca gttccagggc gctgttgatc actttgctct tgtccagccg
     1321 gctcatgtcg acattctttg ccaaaatgat gagacagcac aataaccagc acgttgccca
     1381 ggagctgtag gaaaaagaag aaggcatgaa catggttagc agaggctcta gagccgccgg
     1441 tcacacgcca gaagccgaac cccgccctgc cccgtccccc ccgaaggcag ccgtcccccc
     1501 gcggacagcc ccgaggctgg agagggagaa ggggacggcg gcgcggcgac gcacgaaggc
     1561 cctccccgcc catttccttc ctgccggcgc cgcaccgctt cgccccgcgc ccgctagagg
     1621 gggtgcggcg gcgcctccca gatttcggct ccgcacagat ttgggacaaa ggaagtccct
     1681 gcgccctctc gcacgattac cataaaaggc aatggctgcg gctcgccgcg cctcgacagc
     1741 cgccggcgct ccgggggccg ccgcgcccct cccccgagcc ctccccggcc cgaggcggcc
     1801 ccgccccgcc cggcaccccc acctgccgcc accccccgcc cggcacggcg agccccgcgc
     1861 cacgccccgt acggagcccc gcacccgaag ccgggccgtg ctcagcaact cggggagggg
     1921 ggtgcagggg gggttgcagc ccgaccgacg cgcccacacc ccctgctcac ccccccacgc
     1981 acacaccccg cacgcagcct ttgttcccct cgcagccccc cccgcaccgc ggggcaccgc
     2041 ccccggccgc gctcccctcg cgcacactgc ggagcgcaca aagccccgcg ccgcgcccgc
     2101 agcgctcaca gccgccgggc agcgcggagc cgcacgcggc gctccccacg cacacacaca
     2161 cgcacgcacc ccccgagccg ctccccccgc acaaagggcc ctcccggagc ccctcaaggc
     2221 tttcacgcag ccacagaaaa gaaacaagcc gtcattaaac caagcgctaa ttacagcccg
     2281 gaggagaagg gccgtcccgc ccgctcacct gtgggagtaa cgcggtcagt cagagccggg
     2341 gcgggcggcg cgaggcggcg gcggagcggg gcacggggcg aaggcagcgc gcagcgactc
     2401 ccgcccgccg cgcgcttcgc tttttatagg gccgccgccg ccgccgcctc gccataaaag
     2461 gaaactttcg gagcgcgccg ctctgattgg ctgccgccgc acctctccgc ctcgccccgc
     2521 cccgcccctc gccccgcccc gccccgcctg gcgcgcgccc cccccccccc cccgccccca
     2581 tcgctgcaca aaataattaa aaaataaata aatacaaaat tgggggtggg gagggggggg
     2641 agatggggag agtgaagcag aacgtggggc tcacctcgac catggtaata gcgatgacta
     2701 atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc
     2761 aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt
     2821 gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag
     2881 tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt
     2941 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg
     3001 ggctatgaac taatgacccc gtaattgatt actattaata actagtcaat aatcaatgtc
     3061 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta
     3121 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
     3181 cccagtgcaa gtgcaggtgc cagaacattt ctctgagctc acggggacag ccccccccca
     3241 aagcccccag ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg
     3301 ggctccgctc cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc
     3361 ccgggcacgg ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg
     3421 agcctgcaga cacctggggg gatacgggga aaaagcttta ggctgaaaga gagatttaga
     3481 atgacagaat catagaacgg cctgggttgc aaaggagcac agtgctcatc cagatccaac
     3541 cccctgctat gtgcagggtc atcaaccagc agcccaggct gcccagagcc acatccagcc
     3601 tggccttgaa tgcctgcagg gatggggcat ccacagcctc cttgggcaac ctgttcagtg
     3661 cgtcaccacc ctctggggga aaaactgcct cctcatatcc aacccaaacc tcccctgtct
     3721 cagtgtaaag ccattccccc ttgtcctatc aagggggagt ttgctgtgac attgttggtc
     3781 tggggtgaca catgtttgcc aattcagtgc atcacggaga ggcagatctt ggggataagg
     3841 aagtgcagga cagcatggac gtgggacatg caggtgttga gggctctggg acactctcca
     3901 agtcacagcg ttcagaacag ccttaaggat aagaagatag gatagaagga caaagagcaa
     3961 gttaaaaccc agcatggaga ggagcacaaa aaggccacag acactgctgg tccctgtgtc
     4021 tgagcctgca tgtttgatgg tgtctggatg caagcagaag gggtggaaga gcttgcctgg
     4081 agagatacag ctgggtcagt aggactggga caggcagctg gagaattgcc atgtagatgt
     4141 tcatacaatc gtcaaatcat gaaggctgga aaagccctcc aagatcccca agaccaaccc
     4201 caacccaccc accgtgccca ctggccatgt ccctcagtgc cacatcccca cagttcttca
     4261 tcacctccag ggacggtgac ccccccacct ccgtgggcag ctgtgccact gcagcaccgc
     4321 tctttggaga aggtaaatct tgctaaatcc agcccgaccc tcccctggca caacgtaagg
     4381 ccattatctc tcatccaact ccaggacgga gtcagtgaga atattataac ttcgtatagc
     4441 atacattata cgaagttatg ggcagagcgc acatcgccca cagtccccga gaagttgggg
     4501 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
     4561 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
     4621 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
     4681 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
     4741 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
     4801 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
     4861 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
     4921 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
     4981 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
     5041 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
     5101 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
     5161 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     5221 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     5281 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     5341 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     5401 tagttctcgc gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     5461 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     5521 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     5581 gtggttcaaa gtttttttct tccatttcag gtgtcgtgat ttgcggccgc gccgccacca
     5641 tgacagagta caaaccaact gtacgccttg caacaagaga cgacgtacct cgggccgtca
     5701 gaacactggc tgctgctttc gccgactacc ctgctacacg gcacacggtt gacccagacc
     5761 ggcacattga acgggtgaca gagcttcaag agcttttcct cacccgagtg gggttggaca
     5821 ttgggaaggt ttgggtcgcg gatgacggcg ctgccgtggc ggtatggacg acaccagaga
     5881 gtgttgaggc aggtgctgta ttcgctgaga ttggtccgag gatggcggaa ctgagtgggt
     5941 cccgactcgc agctcagcaa caaatggagg gccttctggc cccccatcgg ccgaaagagc
     6001 ctgcatggtt tttggcaaca gttggcgtat ccccggacca ccagggcaag ggactgggtt
     6061 ctgctgttgt cctgccaggg gttgaggccg ctgaaagggc aggtgtcccg gcatttcttg
     6121 aaacctctgc accaaggaac cttccgttct acgagcgact tggtttcacc gttactgcgg
     6181 atgttgaagt accagaagga ccgaggactt ggtgtatgac aagaaagccg ggcgcatagt
     6241 aagaattctg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     6301 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     6361 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     6421 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggataacttc
     6481 gtatagcata cattatacga agttatttgg tatggctttt tccccgtatc cccccaggtg
     6541 tctgcaggct caaagagcag cgagaagcgt tcagaggaaa gcgatcccgt gccaccttcc
     6601 ccgtgcccgg gctgtccccg cacgctgccg gctcggggat gcggggggag cgccggaccg
     6661 gagcggagcc ccgggcggct cgctgctgcc ccctagcggg ggagggacgt aattacatcc
     6721 ctgggggctt tggggggggg ctgtccctga ttctataaca agaaaatata tatataataa
     6781 gttatcacgt aagtagaaca tgaaataaca atataattat cgtatgagtt aaatcttaaa
     6841 agtcacgtaa aagataatca tgcgtcattt tgactcacgc ggtcgttata gttcaaaatc
     6901 agtgacactt accgcattga caagcacgcc tcacgggagc tccaagcggc gactgagatg
     6961 tcctaaatgc acagcgacgg attcgcgcta tttagaaaga gagagcaata tttcaagaat
     7021 gcatgcgtca attttacgca gactatcttt ctagggttaa tctagctgca tcaggatcat
     7081 atcgtcgggt cttttttccg gctcagtcat cgcccaagct ggcgctatct gggcatcggg
     7141 gaggaagaag cccgtgcctt ttcccgcgag gttgaagcgg catggaaaga gtttgccgag
     7201 gatgactgct gctgcattga cgttgagcga aaacgcacgt ttaccatgat gattcgggaa
     7261 ggtgtggcca tgcacgcctt taacggtgaa ctgttcgttc aggccacctg ggataccagt
     7321 tcgtcgcggc ttttccggac acagttccgg atggtcagcc cgaagcgcat cagcaacccg
     7381 aacaataccg gcgacagccg gaactgccgt gccggtgtgc agattaatga cagcggtgcg
     7441 gcgctgggat attacgtcag cgaggacggg tatcctggct ggatgccgca gaaatggaca
     7501 tggatacccc gtgagttacc cggcgggcgc gcttggcgta atcatggtca tagctgtttc
     7561 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     7621 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     7681 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     7741 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     7801 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7861 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7921 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7981 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     8041 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     8101 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     8161 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     8221 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     8281 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     8341 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     8401 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     8461 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     8521 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     8581 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     8641 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     8701 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     8761 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     8821 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     8881 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     8941 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     9001 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     9061 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     9121 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     9181 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     9241 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     9301 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     9361 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     9421 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     9481 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     9541 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     9601 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     9661 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     9721 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     9781 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     9841 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     9901 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     9961 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
    10021 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
    10081 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
    10141 gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
    10201 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
    10261 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc
    10321 ctcgttcatt cacgtttttg aacccgtgga ggacgggcag actcgcggtg caaatgtgtt
    10381 ttacagcgtg atggagcaga tgaagatgct cgacacgctg cagaacacgc agctagatta
    10441 accctagaaa gataatcata ttgtgacgta cgttaaagat aatcatgtgt aaaattgacg
    10501 catgtgtttt atcggtctgt atatcgaggt ttatttatta atttgaatag atattaagtt
    10561 ttattatatt tacacttaca tactaataat aaattcaaca aacaatttat ttatgtttat
    10621 ttatttatta aaaaaaacaa aaactcaaaa tttcttctat aaagtaacaa aacttttatg
    10681 agggacagcc cccccccaaa gcccccaggg atgtaattac gtccctcccc cgctaggggg
    10741 cagcagcgag ccgcccgggg ctccgctccg gtccggcgct ccccccgcat ccccgagccg
    10801 gcagcgtgcg gggacagccc gggcacgggg aaggtggcac gggatcgctt tcctctgaac
    10861 gcttctcgct gctctttgag cctgcagaca cctgggggga tacggggaaa aggcctccac
    10921 ggccttaatt aagggaggtg gtccctgcag ttacgccaat gataaccccc gccagaaaaa
    10981 tcttagtagc cttccctttt tgttttccgt gccccaactc ggcggattga ctcggcccct
    11041 tccggaaaca cccgaatcaa cttctagtca aattattgtt cacgccgcaa tgacccaccc
    11101 ctggcccgcg tctgtggaac tgacccctgg tgtacaggag agttcgctgc tgaaagtggt
    11161 cccaaagggg tactagtttt taagctccca actccccctc ccccagcgtc tggaggattc
    11221 cacaccctcg caccgcaggg gcgaggaagt gggcggagtc cggttttggc gccagccgct
    11281 gaggctgcca agcagaaaag ccaccgctga ggagactccg gtcactgtcc tcgccccgcc
    11341 tcccccttcc ctccccttgg ggaccaccgg gcgccacgcc gcgaacggta agtgccgcgg
    11401 tcgtcggcgc ctccgccctc cccctagggc cccaattccc agcgggcgcg gcgcgcggcc
    11461 cctccccccg ccgggcgcgc gcccgctgcc ccgcccttcg tggccgcccg gcgtgggcgg
    11521 tgccacccct ccccccggcg gccccgcgcg cagctcccgg ctccctcccc cttcggatgt
    11581 ggcttgagct gtaggcgcgg agggtaccag tttactccct atcagtgata gagaacgtat
    11641 gaagagttta ctccctatca gtgatagaga acgtatgcag actttactcc ctatcagtga
    11701 tagagaacgt ataaggagtt tactccctat cagtgataga gaacgtatga ccagtttact
    11761 ccctatcagt gatagagaac gtatctacag tttactccct atcagtgata gagaacgtat
    11821 atccagttta ctccctatca gtgatagaga acgtataagc tttaggcgtg tacggtgggc
    11881 gcctataaaa gcagagctcg tttagtgaac cgtcagatcg cctggagcaa ttccacaaca
    11941 cttttgtctt ataccaactt tccgtaccac ttcctaccct cgtaaaggat ccgccgccac
    12001 catgtttgtg aagtctgaaa cactggaact taaagaagag gaagatgtct tggttttgct
    12061 gggctctgca gcaccagcgc tcgccgcttt gacgcctttg agtagttccg ctgatgagga
    12121 agaagaagaa gaacccggag catccggtgg tgctcgccgc caaagaggag cagaagctgg
    12181 acaaggagct cgcggcggtg tcgcagccgg ggcagaaggt tgtaggccag ccaggttgct
    12241 cgggttggtg catgactgta agaggagacc gagcagggca agggctgtga gtagaggtgc
    12301 taaaactgca gaaactgtcc aaagaattaa gaagactaga cgacttaaag ctaataatag
    12361 ggaacgcaat aggatgcata atctgaatgc cgccctcgat gcccttagag aagtactgcc
    12421 aacttttcct gaagatgcaa aactgacaaa gattgaaact ctccgttttg cacataatta
    12481 tatttgggct ttgactgaaa cactccggct tgcagaccat tgtggtggtg gcggaggagg
    12541 tcttccaggc gccctgttta gcgaagccgt cctcttggcc cctggtggag cgtcagctgc
    12601 tctctcttcc tcaggcgatg caccggcccc agcttcaacc tggtcatgta caaatgcccc
    12661 agccccaagc agtagcgtta gctctaactc aacagcccct tatagttgta ccctcgcacc
    12721 ggcagcccct gcgggtagtg atatggatta ctggcaacct cctccaccgg ataaacatcg
    12781 ttacgcccca catttgccaa tcgcgcgaga ctgcattctg taagttaacg tttaaactcg
    12841 cgaatcgatc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
    12901 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
    12961 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
    13021 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
    13081 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
    13141 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
    13201 ttaatgaatc ggccaacgc
//
