LOCUS       sequence-472335-       10618 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor construct for introduction of NGN2 into AAVS1 safe harbor site
            and iPSC differentiation to cortical neuron.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10618)
  AUTHORS   Fernandopulle MS, Prestil R, Grunseich C, Wang C, Gan L, Ward ME
  TITLE     Transcription Factor-Mediated Differentiation of Human iPSCs into 
            Neurons.
  JOURNAL   Curr Protoc Cell Biol. 2018 Jun;79(1):e51. doi: 10.1002/cpcb.51. 
            Epub 2018 May 18.
  PUBMED    29924488
REFERENCE   2  (bases 1 to 10618)
  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: "Curr Protoc
            Cell Biol."; date: "2018-06"; volume: "79(1):e51. doi"; pages: " 
            10.1002/cpcb.51. Epub 2018 May 18"
COMMENT     Sequence Label: pUCM-AAVS1-TO-hNGN2
FEATURES             Location/Qualifiers
     source          1..10618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(54..122,123..914)
                     /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(272..291)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1085..1673
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1574..1593
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1721..2557
                     /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    2590..2608
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2626..2644
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2662..2680
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2698..2716
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2734..2752
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2770..2788
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2806..2824
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2869..2893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     polyA_signal    3832..3953
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3869..3888)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3923..3942
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4002..4018)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4002..4018)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4015..4037)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4026..4042
                     /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(4050..4056,4057..4074,4075..4080))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4095..4116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4233..4250)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4542..4561
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    4560..4615
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     4661..4680
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(4771..5517)
                     /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(5549..5568)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          5576..6592
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(5630..5652)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(6593..6871)
                     /label=chicken beta-actin promoter
     enhancer        6873..7252
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    7258..7482
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     7471..7488
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    7504..7537
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(7813..7832)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     8082..8100
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      8242..8251
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             8289..8557
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(8570..8781)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     polyA_signal    8817..9041
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(9062..9661)
                     /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(9145..9165)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     9642..9661
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(9671..9724)
                     /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    9748..9773
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    9780..9813
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    9815..10618
                     /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 aagggcgaat tcaaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
       61 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      121 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      181 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      241 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
      301 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
      361 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
      421 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
      481 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
      541 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
      601 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
      661 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
      721 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
      781 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
      841 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
      901 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
      961 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1021 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1081 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1141 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1201 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1261 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1321 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1381 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     1441 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1501 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1561 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1621 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     1681 gcaacgcggc ctttttacgg ttcctggcga attcgccctt agagcagagc caggaacccc
     1741 tgtagggaag gggcaggaga gccaggggca tgagatggtg gacgaggaag ggggacaggg
     1801 aagcctgagc gcctctcctg ggcttgccaa ggactcaaac ccagaagccc agagcagggc
     1861 cttagggaag cgggaccctg ctctgggcgg aggaatatgt cccagatagc actggggact
     1921 ctttaaggaa agaaggatgg agaaagagaa agggagtaga ggcggccacg acctggtgaa
     1981 cacctaggac gcaccattct cacaaaggga gttttccaca cggacacccc cctcctcacc
     2041 acagccctgc caggacgggg ctggctactg gccttatctc acaggtaaaa ctgacgcacg
     2101 gaggaacaat ataaattggg gactagaaag gtgaagagcc aaagttagaa ctcaggacca
     2161 acttattctg attttgtttt tccaaactgc ttctcctctt gggaagtgta aggaagctgc
     2221 agcaccagga tcagtgaaac gcaccagaca gccgcgtcag agcagctcag gttctgggag
     2281 agggtagcgc agggtggcca ctgagaaccg ggcaggtcac gcatcccccc cttccctccc
     2341 accccctgcc aagctctccc tcccaggatc ctctctggct ccatcgtaag caaaccttag
     2401 aggttctggc aaggagagag atggctccag gaaatggggg tgtgtcacca gataaggaat
     2461 ctgcctaaca ggaggtgggg gttagaccca atatcaggag actaggaagg aggaggccta
     2521 aggatggggc ttttctgtca ccaatcctgt ccctagtaaa gctgatatct ccaccccaca
     2581 gtggtttact ccctatcagt gatagagaac gtatgaagag tttactccct atcagtgata
     2641 gagaacgtat gcagacttta ctccctatca gtgatagaga acgtataagg agtttactcc
     2701 ctatcagtga tagagaacgt atgaccagtt tactccctat cagtgataga gaacgtatct
     2761 acagtttact ccctatcagt gatagagaac gtatatccag tttactccct atcagtgata
     2821 gagaacgtat aagctttagg cgtgtacggt gggcgcctat aaaagcagag ctcgtttagt
     2881 gaaccgtcag atcgcctgga gcaattccac aacacttttg tcttatacca actttccgta
     2941 ccacttccta ccctcgtaaa cttaaggtta attaagccgc caccatgttc gtcaaatccg
     3001 agaccttgga gttgaaggag gaagaggacg tgttagtgct gctcggatcg gcctcccccg
     3061 ccttggcggc cctgaccccg ctgtcatcca gcgccgacga agaagaggag gaggagccgg
     3121 gcgcgtcagg cggggcgcgt cggcagcgcg gggctgaggc cgggcagggg gcgcggggcg
     3181 gcgtggctgc gggtgcggag ggctgccggc ccgcacggct gctgggtctg gtacacgatt
     3241 gcaaacggcg cccttcccgg gcgcgggccg tctcccgagg cgccaagacg gccgagacgg
     3301 tgcagcgcat caagaagacc cgtagactga aggccaacaa ccgcgagcga aaccgcatgc
     3361 acaacctcaa cgcggcactg gacgcgctgc gcgaggtgct ccccacgttc cccgaggacg
     3421 ccaagctcac caagatcgag accctgcgct tcgcccacaa ctacatctgg gcactcaccg
     3481 agaccctgcg cctggcggat cactgcgggg gcggcggcgg gggcctgccg ggggcgctct
     3541 tctccgaggc agtgttgctg agcccgggag gcgccagcgc cgccctgagc agcagcggag
     3601 acagcccctc gcccgcctcc acgtggagtt gcaccaacag ccccgcgccg tcctcctccg
     3661 tgtcctccaa ttccacctcc ccctacagct gcactttatc gcccgccagc ccggccgggt
     3721 cagacatgga ctattggcag cccccacctc ccgacaagca ccgctatgca cctcacctcc
     3781 ccatagccag ggattgtatc ttgtaagcgg ccgcaatcga tcgcccaccc caacttgttt
     3841 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3901 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     3961 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4021 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4081 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4141 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4201 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4261 gttcggctgc ggcgagcggt atcagctcac tcaaaggacg gccgcacatg taaggaaaat
     4321 tttagggatg ttaaagaaaa aaataacaca aaacaaaata taaaaaaaat ctaacctcaa
     4381 gtcaaggctt ttctatggaa taaggaatgg acagcagggg gctgtttcat atactgatga
     4441 cctctttata gccaaccttt gttcatggca gccagcatat gggcatatgt tgccaaactc
     4501 taaaccaaat actcattctg atgttttaaa tgatttgccc tcccatatgt ccttccgagt
     4561 gagagacaca aaaaattcca acacactatt gcaatgaaaa taaatttcct ttattagcca
     4621 gaagtcagat gctcaagggg cttcatgatg tccccataat ttttggcaga gggaaaaaga
     4681 tctcagtggt atttgtgagc cagggcattg gccacaccag ccaccacctt ctgataggca
     4741 gcctgcacct gaggagtgaa ttcacgcgtt ttagcctggc agcatatcga gatcgaagtc
     4801 atcgagagcg tcagcaggga gcatatccag atcaaaatca tccagggcat cggcgggcag
     4861 catgtccagg tcgaagtcgt ccagggcgtc ggtgggtccg ccgctctcgc acttcagctg
     4921 cttctcgagg ccgcagatga tcagttccag gccgaacagg aaggcgggct cggcgccctg
     4981 tctgtcgaac agctcgatgg cctgcttcag cagggggggc atgctgtcgg tggtgggtgt
     5041 ctctctttcc tctttggcga cctggtgttc ctgttcttcc agcacgcagc ccagggtgaa
     5101 gtggcccacg gcgctcaggg cgtacagggc gttttccagg ctgaagccct gctggcacag
     5161 gaaggccagc tggttttcca gtgtctcgta ctgcttctcg gtgggtctgg tgcccaggtg
     5221 cactttggcg ccgtcccggt ggctcagcag ggcgcatctg tagctcttgg cgttgttccg
     5281 caggaagtcc tgccagctct cgccttccag agggcagctg tgggtgtggt gccggtccag
     5341 catctcgatg ggcagggcgt ccagcagggc ccgcttgttc ttcacgtgcc agtacagggt
     5401 gggctgttcc acgcccagtt tctgggccag cttccgggtg gtcaggccct cgatgcccac
     5461 gccgttcagc agttccaggg cgctgttgat cactttgctc ttgtccagcc ggctcatgtc
     5521 gacattcttt gccaaaatga tgagacagca caataaccag cacgttgccc aggagctgta
     5581 ggaaaaagaa gaaggcatga acatggttag cagaggctct agagccgccg gtcacacgcc
     5641 agaagccgaa ccccgccctg ccccgtcccc cccgaaggca gccgtccccc cgcggacagc
     5701 cccgaggctg gagagggaga aggggacggc ggcgcggcga cgcacgaagg ccctccccgc
     5761 ccatttcctt cctgccggcg ccgcaccgct tcgccccgcg cccgctagag ggggtgcggc
     5821 ggcgcctccc agatttcggc tccgcacaga tttgggacaa aggaagtccc tgcgccctct
     5881 cgcacgatta ccataaaagg caatggctgc ggctcgccgc gcctcgacag ccgccggcgc
     5941 tccgggggcc gccgcgcccc tcccccgagc cctccccggc ccgaggcggc cccgccccgc
     6001 ccggcacccc cacctgccgc caccccccgc ccggcacggc gagccccgcg ccacgccccg
     6061 tacggagccc cgcacccgaa gccgggccgt gctcagcaac tcggggaggg gggtgcaggg
     6121 ggggttgcag cccgaccgac gcgcccacac cccctgctca cccccccacg cacacacccc
     6181 gcacgcagcc tttgttcccc tcgcagcccc ccccgcaccg cggggcaccg cccccggccg
     6241 cgctcccctc gcgcacactg cggagcgcac aaagccccgc gccgcgcccg cagcgctcac
     6301 agccgccggg cagcgcggag ccgcacgcgg cgctccccac gcacacacac acgcacgcac
     6361 cccccgagcc gctccccccg cacaaagggc cctcccggag cccctcaagg ctttcacgca
     6421 gccacagaaa agaaacaagc cgtcattaaa ccaagcgcta attacagccc ggaggagaag
     6481 ggccgtcccg cccgctcacc tgtgggagta acgcggtcag tcagagccgg ggcgggcggc
     6541 gcgaggcggc ggcggagcgg ggcacggggc gaaggcagcg cgcagcgact cccgcccgcc
     6601 gcgcgcttcg ctttttatag ggccgccgcc gccgccgcct cgccataaaa ggaaactttc
     6661 ggagcgcgcc gctctgattg gctgccgccg cacctctccg cctcgccccg ccccgcccct
     6721 cgccccgccc cgccccgcct ggcgcgcgcc cccccccccc ccccgccccc atcgctgcac
     6781 aaaataatta aaaaataaat aaatacaaaa ttgggggtgg ggaggggggg ggagatgggg
     6841 agagtgaagc agaacgtggg gctcacctcg accatggtaa tagcgatgac taatacgtag
     6901 atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc
     6961 catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact
     7021 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
     7081 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt
     7141 cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga
     7201 actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg tcgagaccca
     7261 tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt
     7321 cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt
     7381 cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg
     7441 gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct gatcagcggg
     7501 tttataactt cgtataatgt atgctatacg aagttatggt accttactcg tccatgccgc
     7561 cggtggagtg gcggccctcg gcgcgttcgt actgttccac gatggtgtag tcctcgttgt
     7621 gggaggtgat gtccaacttg atgttgacgt tgtaggcgcc gggcagctgc acgggcttct
     7681 tggccttgta ggtggtcttg acctcagcgt cgtagtggcc gccgtccttc agcttcagcc
     7741 tctgcttgat ctcgcccttc agggcgccgt cctcggggta catccgctcg gaggaggcct
     7801 cccagcccat ggtcttcttc tgcattacgg ggccgtcgga ggggaagttg gtgccgcgca
     7861 gcttcacctt gtagatgaac tcgccgtcct gcagggagga gtcctgggtc acggtcacca
     7921 cgccgccgtc ctcgaagttc atcacgcgct cccacttgaa gccctcgggg aaggacagct
     7981 tcaagtagtc ggggatgtcg gcggggtgct tcacgtaggc cttggagccg tacatgaact
     8041 gaggggacag gatgtcccag gcgaagggca gggggccacc cttggtcacc ttcagcttgg
     8101 cggtctgggt gccctcgtag gggcggccct cgccctcgcc ctcgatctcg aactcgtggc
     8161 cgttcacgga gccctccatg tgcaccttga agcgcatgaa ctccttgatg atggccatgt
     8221 tatcctcctc gcccttgctc accatggtgg cgaccggtag cgctagcgta ggcgccggtc
     8281 acagcttgga tctgtaacgg cgcagaacag aaaacgaaac aaagacgtag agttgagcaa
     8341 gcagggtcag gcaaagcgtg gagagccggc tgagtctagg taggctccaa gggagcgccg
     8401 gacaaaggcc cggtctcgac ctgagcttta aacttaccta gacggcggac gcagttcagg
     8461 aggcaccaca ggcgggaggc ggcagaacgc gactcaaccg gcgtggatgg cggcctcagg
     8521 tagggcggcg ggcgcgtgaa ggagagatgc gagcccctcg aagcttcagc tgtgttctgg
     8581 cggcaaaccc gttgcgaaaa agaacgttca cggcgactac tgcacttata tacggttctc
     8641 ccccaccctc gggaaaaagg cggagccagt acacgacatc actttcccag tttaccccgc
     8701 gccaccttct ctaggcaccg gttcaattgc cgacccctcc ccccaacttc tcggggactg
     8761 tgggcgatgt gcgctctgcc cactgacggg caccggagcg atcgcagatc atgcatccat
     8821 agagcccacc gcatccccag catgcctgct attgtcttcc caatcctccc ccttgctgtc
     8881 ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc
     8941 ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg
     9001 ggaggggcaa acaacagatg gctggcaact agaaggcaca ggtttaaacg ggccctctag
     9061 atcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc acctcgacgt
     9121 cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc gcggaggtct
     9181 ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac tccggggagc acgacggcgc
     9241 tgcccagacc cttgccctgg tggtcgggcg agacgccgac ggtggccagg aaccacgcgg
     9301 gctccttggg ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg gccagccggg
     9361 aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa caccgccccc gcttcgacgc
     9421 tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac accttgccga
     9481 tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc tcgatgtggc
     9541 ggtccggatc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc
     9601 gtacggccct ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca
     9661 tctcgagcct agggccggga ttctcctcca cgtcaccgca tgttagaaga cttcctctgc
     9721 cctctccgct gccagatctc tcgaggccct gtgggaggaa gagaagaggt cagaagctta
     9781 taacttcgta taatgtatgc tatacgaagt tattgcccca ctgtggggtg gaggggacag
     9841 ataaaagtac ccagaaccag agccacatta accggccctg ggaatataag gtggtcccag
     9901 ctcggggaca caggatccct ggaggcagca aacatgctgt cctgaagtgg acataggggc
     9961 ccgggttgga ggaagaagac tagctgagct ctcggacccc tggaagatgc catgacaggg
    10021 ggctggaaga gctagcacag actagagagg taaggggggt aggggagctg cccaaatgaa
    10081 aggagtgaga ggtgacccga atccacagga gaacggggtg tccaggcaaa gaaagcaaga
    10141 ggatggagag gtggctaaag ccagggagac ggggtacttt ggggttgtcc agaaaaacgg
    10201 tgatgatgca ggcctacaag aaggggaggc gggacgcaag ggagacatcc gtcggagaag
    10261 gccatcctaa gaaacgagag atggcacagg ccccagaagg agaaggaaaa gggaacccag
    10321 cgagtgaaga cggcatgggg ttgggtgagg gaggagagat gcccggagag gacccagaca
    10381 cggggaggat ccgctcagag gacatcacgt ggtgcagcgc cgagaaggaa gtgctccgga
    10441 aagagcatcc ttgggcagca acacagcaga gagcaagggg aagagggagt ggaggaagac
    10501 ggaacctgaa ggaggcggca gggaaggatc tgggccagcc gtagaggtga cccaggccac
    10561 aagctgcaga cagaaagcgg cacaggccca ggggagagaa tgctggtcag agaaagca
//
