LOCUS       sequence-207872-       10709 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Donor construct for introduction of NGN2 into AAVS1 safe harbor site
            and iPSC differentiation to cortical neuron, TALEN cut site outside 
            of homology arm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10709)
  TITLE     A comprehensive guide for transcription-factor mediated 
            differentiation of human iPSC into neurons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10709)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: H52-pUCM-hNGN2-inv
FEATURES             Location/Qualifiers
     source          1..10709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    14..32
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    50..68
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    86..104
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    122..140
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    158..176
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    194..212
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    230..248
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     293..317
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     polyA_signal    1256..1377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1293..1312)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1347..1366
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1426..1442)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1426..1442)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1439..1461)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1450..1466
                     /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(1474..1480,1481..1498,1499..1504))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1519..1540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1657..1674)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1966..1985
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1984..2039
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     2085..2104
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(2194..2940)
                     /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(2972..2991)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          2999..4016
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(3053..3075)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(4018..4295)
                     /label=chicken beta-actin promoter
     enhancer        4297..4676
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    4682..4906
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4895..4912
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    4928..4961
                     /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(5237..5256)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5506..5524
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      5666..5675
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             5713..5981
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(5994..6205)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     polyA_signal    6241..6465
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(6486..7085)
                     /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(6569..6589)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7066..7085
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(7095..7148)
                     /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    7172..7197
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    7204..7237
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    7239..8042
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     CDS             join(8187..8255,8256..9047)
                     /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(8405..8424)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9218..9806
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9707..9726
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    9854..10690
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
ORIGIN
        1 cacagtggtt tactccctat cagtgataga gaacgtatga agagtttact ccctatcagt
       61 gatagagaac gtatgcagac tttactccct atcagtgata gagaacgtat aaggagttta
      121 ctccctatca gtgatagaga acgtatgacc agtttactcc ctatcagtga tagagaacgt
      181 atctacagtt tactccctat cagtgataga gaacgtatat ccagtttact ccctatcagt
      241 gatagagaac gtataagctt taggcgtgta cggtgggcgc ctataaaagc agagctcgtt
      301 tagtgaaccg tcagatcgcc tggagcaatt ccacaacact tttgtcttat accaactttc
      361 cgtaccactt cctaccctcg taaacttaag gttaattaag ccgccaccat gttcgtcaaa
      421 tccgagacct tggagttgaa ggaggaagag gacgtgttag tgctgctcgg atcggcctcc
      481 cccgccttgg cggccctgac cccgctgtca tccagcgccg acgaagaaga ggaggaggag
      541 ccgggcgcgt caggcggggc gcgtcggcag cgcggggctg aggccgggca gggggcgcgg
      601 ggcggcgtgg ctgcgggtgc ggagggctgc cggcccgcac ggctgctggg tctggtacac
      661 gattgcaaac ggcgcccttc ccgggcgcgg gccgtctccc gaggcgccaa gacggccgag
      721 acggtgcagc gcatcaagaa gacccgtaga ctgaaggcca acaaccgcga gcgaaaccgc
      781 atgcacaacc tcaacgcggc actggacgcg ctgcgcgagg tgctccccac gttccccgag
      841 gacgccaagc tcaccaagat cgagaccctg cgcttcgccc acaactacat ctgggcactc
      901 accgagaccc tgcgcctggc ggatcactgc gggggcggcg gcgggggcct gccgggggcg
      961 ctcttctccg aggcagtgtt gctgagcccg ggaggcgcca gcgccgccct gagcagcagc
     1021 ggagacagcc cctcgcccgc ctccacgtgg agttgcacca acagccccgc gccgtcctcc
     1081 tccgtgtcct ccaattccac ctccccctac agctgcactt tatcgcccgc cagcccggcc
     1141 gggtcagaca tggactattg gcagccccca cctcccgaca agcaccgcta tgcacctcac
     1201 ctccccatag ccagggattg tatcttgtaa gcggccgcaa tcgatcgccc accccaactt
     1261 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     1321 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     1381 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     1441 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     1501 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     1561 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     1621 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     1681 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gacggccgca catgtaagga
     1741 aaattttagg gatgttaaag aaaaaaataa cacaaaacaa aatataaaaa aaatctaacc
     1801 tcaagtcaag gcttttctat ggaataagga atggacagca gggggctgtt tcatatactg
     1861 atgacctctt tatagccaac ctttgttcat ggcagccagc atatgggcat atgttgccaa
     1921 actctaaacc aaatactcat tctgatgttt taaatgattt gccctcccat atgtccttcc
     1981 gagtgagaga cacaaaaaat tccaacacac tattgcaatg aaaataaatt tcctttatta
     2041 gccagaagtc agatgctcaa ggggcttcat gatgtcccca taatttttgg cagagggaaa
     2101 aagatctcag tggtatttgt gagccagggc attggccaca ccagccacca ccttctgata
     2161 ggcagcctgc acctgaggag tgaattcacg cgtttagcct ggcagcatat cgagatcgaa
     2221 gtcatcgaga gcgtcagcag ggagcatatc cagatcaaaa tcatccaggg catcggcggg
     2281 cagcatgtcc aggtcgaagt cgtccagggc gtcggtgggt ccgccgctct cgcacttcag
     2341 ctgcttctcg aggccgcaga tgatcagttc caggccgaac aggaaggcgg gctcggcgcc
     2401 ctgtctgtcg aacagctcga tggcctgctt cagcaggggg ggcatgctgt cggtggtggg
     2461 tgtctctctt tcctctttgg cgacctggtg ttcctgttct tccagcacgc agcccagggt
     2521 gaagtggccc acggcgctca gggcgtacag ggcgttttcc aggctgaagc cctgctggca
     2581 caggaaggcc agctggtttt ccagtgtctc gtactgcttc tcggtgggtc tggtgcccag
     2641 gtgcactttg gcgccgtccc ggtggctcag cagggcgcat ctgtagctct tggcgttgtt
     2701 ccgcaggaag tcctgccagc tctcgccttc cagagggcag ctgtgggtgt ggtgccggtc
     2761 cagcatctcg atgggcaggg cgtccagcag ggcccgcttg ttcttcacgt gccagtacag
     2821 ggtgggctgt tccacgccca gtttctgggc cagcttccgg gtggtcaggc cctcgatgcc
     2881 cacgccgttc agcagttcca gggcgctgtt gatcactttg ctcttgtcca gccggctcat
     2941 gtcgacattc tttgccaaaa tgatgagaca gcacaataac cagcacgttg cccaggagct
     3001 gtaggaaaaa gaagaaggca tgaacatggt tagcagaggc tctagagccg ccggtcacac
     3061 gccagaagcc gaaccccgcc ctgccccgtc ccccccgaag gcagccgtcc ccccgcggac
     3121 agccccgagg ctggagaggg agaaggggac ggcggcgcgg cgacgcacga aggccctccc
     3181 cgcccatttc cttcctgccg gcgccgcacc gcttcgcccc gcgcccgcta gagggggtgc
     3241 ggcggcgcct cccagatttc ggctccgcac agatttggga caaaggaagt ccctgcgccc
     3301 tctcgcacga ttaccataaa aggcaatggc tgcggctcgc cgcgcctcga cagccgccgg
     3361 cgctccgggg gccgccgcgc ccctcccccg agccctcccc ggcccgaggc ggccccgccc
     3421 cgcccggcac ccccacctgc cgccaccccc cgcccggcac ggcgagcccc gcgccacgcc
     3481 ccgtacggag ccccgcaccc gaagccgggc cgtgctcagc aactcgggga ggggggtgca
     3541 gggggggttg cagcccgacc gacgcgccca caccccctgc tcaccccccc acgcacacac
     3601 cccgcacgca gcctttgttc ccctcgcagc cccccccgca ccgcggggca ccgcccccgg
     3661 ccgcgctccc ctcgcgcaca ctgcggagcg cacaaagccc cgcgccgcgc ccgcagcgct
     3721 cacagccgcc gggcagcgcg gagccgcacg cggcgctccc cacgcacaca cacacgcacg
     3781 caccccccga gccgctcccc ccgcacaaag ggccctcccg gagcccctca aggctttcac
     3841 gcagccacag aaaagaaaca agccgtcatt aaaccaagcg ctaattacag cccggaggag
     3901 aagggccgtc ccgcccgctc acctgtggga gtaacgcggt cagtcagagc cggggcgggc
     3961 ggcgcgaggc ggcggcggag cggggcacgg ggcgaaggca gcgtcgcagc gactccccgc
     4021 ccgccgcgcg cttcgctttt tatagggccg ccgccgccgc cgcctcgcca taaaaggaaa
     4081 ctttcggagc gcgccgctct gattggctgc cgccgcacct ctccgcctcg ccccgccccg
     4141 cccctcgccc cgccccgccc cgcctggcgc gcgccccccc cccccccccg cccccatcgc
     4201 tgcacaaaat aattaaaaaa taaataaata caaaattggg ggtggggagg ggggggagat
     4261 ggggagagtg aagcagaacg tggggctcac ctcgaccatg gtaatagcga tgactaatac
     4321 gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc
     4381 gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg
     4441 tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc
     4501 tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg
     4561 cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct
     4621 atgaactaat gaccccgtaa ttgattacta ttaataacta gtcaataatc aatgtcgaga
     4681 cccatagagc ccaccgcatc cccagcatgc ctgctattgt cttcccaatc ctcccccttg
     4741 ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca atgcgatgca
     4801 atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt caaggaaggc
     4861 acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga ggctgatcag
     4921 cgggtttata acttcgtata atgtatgcta tacgaagtta tggtacctta ctcgtccatg
     4981 ccgccggtgg agtggcggcc ctcggcgcgt tcgtactgtt ccacgatggt gtagtcctcg
     5041 ttgtgggagg tgatgtccaa cttgatgttg acgttgtagg cgccgggcag ctgcacgggc
     5101 ttcttggcct tgtaggtggt cttgacctca gcgtcgtagt ggccgccgtc cttcagcttc
     5161 agcctctgct tgatctcgcc cttcagggcg ccgtcctcgg ggtacatccg ctcggaggag
     5221 gcctcccagc ccatggtctt cttctgcatt acggggccgt cggaggggaa gttggtgccg
     5281 cgcagcttca ccttgtagat gaactcgccg tcctgcaggg aggagtcctg ggtcacggtc
     5341 accacgccgc cgtcctcgaa gttcatcacg cgctcccact tgaagccctc ggggaaggac
     5401 agcttcaagt agtcggggat gtcggcgggg tgcttcacgt aggccttgga gccgtacatg
     5461 aactgagggg acaggatgtc ccaggcgaag ggcagggggc cacccttggt caccttcagc
     5521 ttggcggtct gggtgccctc gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg
     5581 tggccgttca cggagccctc catgtgcacc ttgaagcgca tgaactcctt gatgatggcc
     5641 atgttatcct cctcgccctt gctcaccatg gtggcgaccg gtagcgctag cgtaggcgcc
     5701 ggtcacagct tggatctgta acggcgcaga acagaaaacg aaacaaagac gtagagttga
     5761 gcaagcaggg tcaggcaaag cgtggagagc cggctgagtc taggtaggct ccaagggagc
     5821 gccggacaaa ggcccggtct cgacctgagc tttaaactta cctagacggc ggacgcagtt
     5881 caggaggcac cacaggcggg aggcggcaga acgcgactca accggcgtgg atggcggcct
     5941 caggtagggc ggcgggcgcg tgaaggagag atgcgagccc ctcgaagctt cagctgtgtt
     6001 ctggcggcaa acccgttgcg aaaaagaacg ttcacggcga ctactgcact tatatacggt
     6061 tctcccccac cctcgggaaa aaggcggagc cagtacacga catcactttc ccagtttacc
     6121 ccgcgccacc ttctctaggc accggttcaa ttgccgaccc ctccccccaa cttctcgggg
     6181 actgtgggcg atgtgcgctc tgcccactga cgggcaccgg agcgatcgca gatcatgcat
     6241 ccatagagcc caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc tcccccttgc
     6301 tgtcctgccc caccccaccc cccagaatag aatgacacct actcagacaa tgcgatgcaa
     6361 tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca
     6421 cgggggaggg gcaaacaaca gatggctggc aactagaagg cacaggttta aacgggccct
     6481 ctagatcagg caccgggctt gcgggtcatg caccaggtgc gcggtccttc gggcacctcg
     6541 acgtcggcgg tgacggtgaa gccgagccgc tcgtagaagg ggaggttgcg gggcgcggag
     6601 gtctccagga aggcgggcac cccggcgcgc tcggccgcct ccactccggg gagcacgacg
     6661 gcgctgccca gacccttgcc ctggtggtcg ggcgagacgc cgacggtggc caggaaccac
     6721 gcgggctcct tgggccggtg cggcgccagg aggccttcca tctgttgctg cgcggccagc
     6781 cgggaaccgc tcaactcggc catgcgcggg ccgatctcgg cgaacaccgc ccccgcttcg
     6841 acgctctccg gcgtggtcca gaccgccacc gcggcgccgt cgtccgcgac ccacaccttg
     6901 ccgatgtcga gcccgacgcg cgtgaggaag agttcttgca gctcggtgac ccgctcgatg
     6961 tggcggtccg gatcgacggt gtggcgcgtg gcggggtagt cggcgaacgc ggcggcgagg
     7021 gtgcgtacgg ccctggggac gtcgtcgcgg gtggcgaggc gcaccgtggg cttgtactcg
     7081 gtcatctcga gcctagggcc gggattctcc tccacgtcac cgcatgttag aagacttcct
     7141 ctgccctctc cgctgccaga tctctcgagg ccctgtggga ggaagagaag aggtcagaag
     7201 cttataactt cgtataatgt atgctatacg aagttattgc cccactgtgg ggtggagggg
     7261 acagataaaa gtacccagaa ccagagccac attaaccggc cctgggaata taaggtggtc
     7321 ccagctcggg gacacaggat ccctggaggc agcaaacatg ctgtcctgaa gtggacatag
     7381 gggcccgggt tggaggaaga agactagctg agctctcgga cccctggaag atgccatgac
     7441 agggggctgg aagagctagc acagactaga gaggtaaggg gggtagggga gctgcccaaa
     7501 tgaaaggagt gagaggtgac ccgaatccac aggagaacgg ggtgtccagg caaagaaagc
     7561 aagaggatgg agaggtggct aaagccaggg agacggggta ctttggggtt gtccagaaaa
     7621 acggtgatga tgcaggccta caagaagggg aggcgggacg caagggagac atccgtcgga
     7681 gaaggccatc ctaagaaacg agagatggca caggccccag aaggagaagg aaaagggaac
     7741 ccagcgagtg aagacggcat ggggttgggt gagggaggag agatgcccgg agaggaccca
     7801 gacacgggga ggatccgctc agaggacatc acgtggtgca gcgccgagaa ggaagtgctc
     7861 cggaaagagc atccttgggc agcaacacag cagagagcaa ggggaagagg gagtggagga
     7921 agacggaacc tgaaggaggc ggcagggaag gatctgggcc agccgtagag gtgacccagg
     7981 ccacaagctg cagacagaaa gcggcacagg cccaggggag agaatgctgg tcagagaaag
     8041 caaagggcga attcaaccct gataaatgct tcaataatgg taccgggtac ttttatctgt
     8101 cccctccacc ccacagtgtc cctagtggcc ccaggattgg tgacagaaaa gccccatcct
     8161 taggccaata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     8221 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     8281 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     8341 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     8401 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     8461 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     8521 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     8581 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     8641 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     8701 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     8761 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     8821 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     8881 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     8941 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     9001 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     9061 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     9121 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     9181 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     9241 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     9301 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     9361 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     9421 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     9481 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     9541 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     9601 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     9661 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     9721 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     9781 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     9841 cgaattcgcc cttagagcag agccaggaac ccctgtaggg aaggggcagg agagccaggg
     9901 gcatgagatg gtggacgagg aagggggaca gggaagcctg agcgcctctc ctgggcttgc
     9961 caaggactca aacccagaag cccagagcag ggccttaggg aagcgggacc ctgctctggg
    10021 cggaggaata tgtcccagat agcactgggg actctttaag gaaagaagga tggagaaaga
    10081 gaaagggagt agaggcggcc acgacctggt gaacacctag gacgcaccat tctcacaaag
    10141 ggagttttcc acacggacac ccccctcctc accacagccc tgccaggacg gggctggcta
    10201 ctggccttat ctcacaggta aaactgacgc acggaggaac aatataaatt ggggactaga
    10261 aaggtgaaga gccaaagtta gaactcagga ccaacttatt ctgattttgt ttttccaaac
    10321 tgcttctcct cttgggaagt gtaaggaagc tgcagcacca ggatcagtga aacgcaccag
    10381 acagccgcgt cagagcagct caggttctgg gagagggtag cgcagggtgg ccactgagaa
    10441 ccgggcaggt cacgcatccc ccccttccct cccaccccct gccaagctct ccctcccagg
    10501 atcctctctg gctccatcgt aagcaaacct tagaggttct ggcaaggaga gagatggctc
    10561 caggaaatgg gggtgtgtca ccagataagg aatctgccta acaggaggtg ggggttagac
    10621 ccaatatcag gagactagga aggaggaggc ctaaggatgg ggcttttctg tcaccaatcc
    10681 tgtccctagt aaagctgata tctccaccc
//
