LOCUS       sequence-393690-       14166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor construct for introduction of hNIL factors and SBP-LNGFR for 
            streptavidin bead enrichment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14166)
  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 14166)
  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: 
            CLYBL-(Ef1a-SBP-LNGFR-T2A-mApple)-(CAG-rtTA)-(TRE-hNIL))
FEATURES             Location/Qualifiers
     source          1..14166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        62..441
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          721..1737
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1661..1683
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1745..1764
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1796..2542
                     /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(2633..2652)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2698..2753
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2752..2771)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     3063..3080
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3197..3218
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3233..3238,3239..3256,3257..3263)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3271..3287
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3276..3298
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3295..3311
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3295..3311
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3360..3481
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3371..3390)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3425..3444
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4713..4766)
                     /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"
     regulatory      5038..5047
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5838..5894)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(6814..6838)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    6883..6901
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6919..6937
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6955..6973
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6991..7009
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7027..7045
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7063..7081
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7099..7117
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     rep_origin      complement(8143..8731)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8223..8242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8902..9693,9694..9762))
                     /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     9525..9544
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    complement(10620..10653)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    10654..10679
                     /label=SA
                     /note="splice acceptor site"
     CDS             10703..10756
                     /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             10766..11365
                     /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(10766..10785)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     11262..11282
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    11386..11610
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        11646..11857
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             11870..12138
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      12176..12185
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             12263..12376
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     CDS             13160..13213
                     /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"
     regulatory      13220..13229
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(13943..13976)
                     /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(13992..14009)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    join(13998..14166,1..56)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatgggtct
       61 cgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
      121 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      181 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      241 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      301 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      361 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      421 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
      481 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
      541 atgggggcgg gggggggggg ggggsgcgcg ccgggggggg gggggggggg gggggggggg
      601 gggggggsgr ggsggrgrgg kgsggcggca gccaatcaga gcggcgcgct ccgaaagttt
      661 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      721 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
      781 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      841 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      901 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
      961 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
     1021 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
     1081 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1141 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
     1201 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
     1261 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg kgggggtgcc gggcggggcg
     1321 gggccgccyc gggccggggr gggctcgggg grggggcgcg gcggcccccg gagcgccggc
     1381 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1441 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
     1501 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
     1561 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
     1621 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     1681 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1741 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatg tcgacatgag
     1801 ccggctggac aagagcaaag tgatcaacag cgccctggaa ctgctgaacg gcgtgggcat
     1861 cgagggcctg accacccgga agctggccca gaaactgggc gtggaacagc ccaccctgta
     1921 ctggcacgtg aagaacaagc gggccctgct ggacgccctg cccatcgaga tgctggaccg
     1981 gcaccacacc cacagctgcc ctctggaagg cgagagctgg caggacttcc tgcggaacaa
     2041 cgccaagagc tacagatgcg ccctgctgag ccaccgggac ggcgccaaag tgcacctggg
     2101 caccagaccc accgagaagc agtacgagac actggaaaac cagctggcct tcctgtgcca
     2161 gcagggcttc agcctggaaa acgccctgta cgccctgagc gccgtgggcc acttcaccct
     2221 gggctgcgtg ctggaagaac aggaacacca ggtcgccaaa gaggaaagag agacacccac
     2281 caccgacagc atgccccccc tgctgaagca ggccatcgag ctgttcgaca gacagggcgc
     2341 cgagcccgcc ttcctgttcg gcctggaact gatcatctgc ggcctcgaga agcagctgaa
     2401 gtgcgagagc ggcggaccca ccgacgccct ggacgacttc gacctggaca tgctgcccgc
     2461 cgatgccctg gatgattttg atctggatat gctccctgct gacgctctcg atgacttcga
     2521 tctcgatatg ctgccaggct aaaacgcgtg aattcactcc tcaggtgcag gctgcctatc
     2581 agaaggtggt gactggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     2641 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     2701 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     2761 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     2821 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     2881 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     2941 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     3001 tacatgtgcg gccgtccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3061 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     3121 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     3181 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     3241 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     3301 acagctatga ccatgattac gccaagctct agctagaggt cgacggtata cagacatgat
     3361 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     3421 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3481 tggggtgggc gatcgattca gaactgggcg tgatccactt cgtccagcca ggaggcaggg
     3541 ctggcaggaa aatcggggta gccgccagag ctgcctgtgc tcagatcgct gctagggcca
     3601 ttgccggcca gcactctcat tggaggaggt ccgcctggag cgccagatgg caccaggccc
     3661 aggcttgtat cggggtacac caggctggac agcagaggct gaggtcctgg cagagactga
     3721 ggggcagctg gagaaggtgg cacgccgtaa gggctgccgg gcctcagttc tctatactgc
     3781 tcgggtccgg ccagtccgcc gtgttccaga gagaagttgc ccagggcgcc agaaggtctg
     3841 cccagagcct gggttggctc tcccaggctg ccatacaggc cattggctgg gcccatttcg
     3901 gccaggctag gctcatctgg gaaggacacc tcggcgtcgc tgtcctggcc ttcctgcacg
     3961 ctgtccttgt cgctcttgct gccgcctctg gaccgcttca tgttccggaa gtactggccc
     4021 catctctgcc tgccggcgtc cttcttcagc cgcttctctt tggcccgtct gttctggaac
     4081 cacacttgca ccactctcat gtccaggcct gtctcgctgc tcagctgttc ccgcacatgt
     4141 ctggcgggct tggggctggt gttgtaggcg gacttcaggg tttccagctg cttggcggtg
     4201 atggtggttc tgggtctttt ggcggtggcc tcggcctctc tctgcttggc tgtctcgtag
     4261 tcggctttac acacgagccg gctatcttcc atcaggtaga actcgtcgcc ggtggccagc
     4321 tgtctcttgc acacgacgca ggcgaagcag tgcagatggt acacgaagtc ctgggctctc
     4381 cgcacgacct gtgtaggagg aattcccagc tggcaggcgg cgcacttggt gccgaatctc
     4441 ttgaagaagt cgtccttgca gtacacgctc tcgcctctgc taaagcatct ctcggccagg
     4501 ggggtgtggc agtcgctgca cttcaggcac ttgctgtgcc agtgtctgtc cagggccttc
     4561 aggataaacc ggtccaggat gtgctggtcg cagccggcgc acagggggat ttctcttctc
     4621 aggtcggccc ttctagccag cagggccagc agcagatctc cgccggcaga ttctctggca
     4681 gggcccagtt cgcctctggc ttccatggta ccggggccag ggttctcttc cacgtcgcca
     4741 catgtcagca ggctgcctct gccctcgcca gagcctctgc tgtgcatcaa tgcctcaata
     4801 ggactggcta ccatgctgtt aggtgtatct ggaagttgag aggacattga tgctacttca
     4861 ctgccagtgg aattagagcc cggtcctcct tctgaaaaat tgaccagttg ctgaaaagca
     4921 ggctgatcta tgtcactctg caaggcgaag tcgctcagta ctttccaagg tggctggtaa
     4981 ctttgtactt ccactgggtt agcctgtaag ccaccgtcgt gtctctctgg actggcagcc
     5041 accatgggag ttcctgtcat cccctggata ttagttttgt cattgggctg ctgctgctgg
     5101 agttgcttca tcatgatgct tcgcttcttg tccttgcacc gcttgttttg aaaccagacc
     5161 cggatcacac ggggactgag gcccgtcatc tctaccagtt gctccttcat gagcgcatct
     5221 ggccgcgggt ttgcggcgta gcaggtccgc aaggtgtgca gctgcttctc gttcagcaca
     5281 gtccgcacgc gggtggtctt ctccggctgc ttgtggacgt ggggccgcag ggccggctgc
     5341 ctggcggaga tgggctccgc tgccatttgc agtggccgcg ctggatgcag gggactgagc
     5401 gggtcgccag cgcctagact ggccctctcc accacatcgt ggtctgctcg gcagaagaga
     5461 ccgtcctccc gaagcgcaaa ttcgtcccca gggatgagct ggcggctgca ggccacacag
     5521 cggaaacact cgatgtgata caccttggag cgggcacgca tcacgaagtc gttcttgctg
     5581 aagccgatgc tgcacttggc gcatttgatc ccgtacaacc tgatataatc tcttttacag
     5641 taggttttcc catccctaac aaagcatgta cagctctcgt ccaaatactg attacactcc
     5701 gcacatttca aacatgccgc atgccattcc aaatccggag aaaccctcag aatatactga
     5761 tcgtgaatct gattgccgca accaacacat agggaaatca gacgtttttt ttttggtgga
     5821 tctcccatgt ctcccatagg tccagggttc tcctccacgt ctccagcctg cttcagcagg
     5881 ctgaagttag tagctccgga tcccaagata caatccctgg ctatggggag gtgaggtgca
     5941 tagcggtgct tgtcgggagg tgggggctgc caatagtcca tgtctgaccc ggccgggctg
     6001 gcgggcgata aagtgcagct gtagggggag gtggaattgg aggacacgga ggaggacggc
     6061 gcggggctgt tggtgcaact ccacgtggag gcgggcgagg ggctgtctcc gctgctgctc
     6121 agggcggcgc tggcgcctcc cgggctcagc aacactgcct cggagaagag cgcccccggc
     6181 aggcccccgc cgccgccccc gcagtgatcc gccaggcgca gggtctcggt gagtgcccag
     6241 atgtagttgt gggcgaagcg cagggtctcg atcttggtga gcttggcgtc ctcggggaac
     6301 gtggggagca cctcgcgcag cgcgtccagt gccgcgttga ggttgtgcat gcggtttcgc
     6361 tcgcggttgt tggccttcag tctacgggtc ttcttgatgc gctgcaccgt ctcggccgtc
     6421 ttggcgcctc gggagacggc ccgcgcccgg gaagggcgcc gtttgcaatc gtgtaccaga
     6481 cccagcagcc gtgcgggccg gcagccctcc gcacccgcag ccacgccgcc ccgcgccccc
     6541 tgcccggcct cagccccgcg ctgccgacgc gccccgcctg acgcgcccgg ctcctcctcc
     6601 tcttcttcgt cggcgctgga tgacagcggg gtcagggccg ccaaggcggg ggaggccgat
     6661 ccgagcagca ctaacacgtc ctcttcctcc ttcaactcca aggtctcgga tttgacgaac
     6721 atggtggcgg cttaattaac cttaagttta cgagggtagg aagtggtacg gaaagttggt
     6781 ataagacaaa agtgttgtgg aattgctcca ggcgatctga cggttcacta aacgagctct
     6841 gcttttatag gcgcccaccg tacacgccta aagcttatac gttctctatc actgataggg
     6901 agtaaactgg atatacgttc tctatcactg atagggagta aactgtagat acgttctcta
     6961 tcactgatag ggagtaaact ggtcatacgt tctctatcac tgatagggag taaactcctt
     7021 atacgttctc tatcactgat agggagtaaa gtctgcatac gttctctatc actgataggg
     7081 agtaaactct tcatacgttc tctatcactg atagggagta aaccactgtg gggtggagat
     7141 atctcctcat ccttccaaca taaatatatt ttgggattat atcaacattc aatgttactt
     7201 aaagtgacct tgtaaatatt ttcacaactg agccatgttt gatttgtata cttatgttta
     7261 ctttactgtt tttcctgaag ttaataattg ccttgaattt atttatttct ttaaaaatgt
     7321 ttcattactc aggactgtag tttacattac gattctttgt gttatacagt tgatgggttt
     7381 cttttctttc ttaatttctt taaaaaatag agatggggtc ttactatatt acccaggctg
     7441 gtcttgaagt cctgggctca agtgatcttc ctgtctcagc ctaccaagta gctgagacta
     7501 taggtgcaaa aaagccacta tacctggcta gtttacaggt tttaacaaat gcattatgcc
     7561 acgtatccat tattacagga tcacacaaga tattttcatt accctgaaag catccctgtg
     7621 ttccaccaat tcatcctgcc tccatgagcc gctggcaacc actgatctct atagttttgc
     7681 cttttctaaa atgtcatata attggaatca tacagtctgt agcattttca gactagcttt
     7741 taaaatttgg caatatgcat ttaaggttcc tccttaaatg tgaagggcat agccaatgtg
     7801 gcttgatagc tcatttcttt ttattggtga atatttcatt gtctggatgt tccacagttt
     7861 gtttatccat tcacctattc aatttgcttt ttttctgtgt atctatcact aattcaatac
     7921 tggactctcc aacagagccg tcccgcttcc ctaaggccct gctctgggct tctgggtttg
     7981 agtccttggc aagcccagga gaggcgctca ggcttccctg tcccccttcc tcgtccacca
     8041 tctcatgccc ctggctctcc tgccccttcc ctacaggggt tcctggctct gctctaaggg
     8101 cgaattcgcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     8161 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     8221 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     8281 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     8341 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     8401 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     8461 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     8521 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     8581 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     8641 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     8701 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     8761 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     8821 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     8881 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     8941 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     9001 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     9061 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     9121 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     9181 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     9241 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     9301 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     9361 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     9421 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     9481 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     9541 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     9601 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     9661 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     9721 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     9781 gcaggagaat catgccaatg ggccaatata cattctgacc cacagtttca taataaaata
     9841 aaatggttgt ggttgtaagc cactatgttt cagagtggtt tgttacacag caataaataa
     9901 ctaatatagt aggcatacca tcaagtccaa agtaggtaga gaagaatgta aatagcagag
     9961 caaaacagca tgactggtgg ctgggaggct taaaactggg acaggatcag agtcatgaaa
    10021 gaagtcaaag aaatggttca gaagtaaggc tgagactgac ttacaaaagc tgaaagtccc
    10081 tttaagttgg tgtttggtgc attggcaggg gcaggtatgg tgacttaaaa gagccatgct
    10141 caacaagatc aagcacaaca caatcacggg tcaccccagc agaccttagc gagtctagcc
    10201 atttctttgg tggtggtcac agtcatgctt cagcccagtt tccacttgga caaatggtac
    10261 atattttcaa tgagatgaaa attaagatac aatccatgtg ctcagagagt gatcacagct
    10321 ctgactaaac actgtgcccc aaagtgttga ggaattggga aaacctagct gagttagtgg
    10381 tctcttttct gttacaataa agctcataat gaaaattagc cttctttgtt cttccccaag
    10441 tctttctttc tagacgaaac tactttcaac tgttttaact tccttactgt taacttccat
    10501 atttctagat agtatggtca gactgttatt tcttgacttt taaatgtaag atattatcta
    10561 ctgacttcct tctatgtaag atgaggattg agctctctta cccttctccc agcggccgca
    10621 taacttcgta tagcatacat tatacgaagt tatctgacct cttctcttcc tcccacaggg
    10681 cctcgagaga tctggcagcg gagagggcag aggaagtctt ctaacatgcg gtgacgtgga
    10741 ggagaatccc ggccctaggc tcgagatgac cgagtacaag cccacggtgc gcctcgccac
    10801 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
    10861 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
    10921 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
    10981 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
    11041 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
    11101 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
    11161 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
    11221 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
    11281 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
    11341 catgacccgc aagcccggtg cctgatctag agggcccgtt taaacctgtg ccttctagtt
    11401 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
    11461 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
    11521 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
    11581 ggcatgctgg ggatgcggtg ggctctatgg atgcatgatc tgcgatcgct ccggtgcccg
    11641 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
    11701 ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
    11761 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
    11821 cgttcttttt cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat
    11881 ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt
    11941 tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa
    12001 gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg
    12061 gctctccacg ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt
    12121 tctgcgccgt tacagatcca agctgtgacc ggcgcctacg ctagcgctac cggtcgccac
    12181 catgggctgg tcatgtatca ttctgtttct ggtcgcaacc gcaactggag tgcattcaca
    12241 ggtgcagctc gaggggtcag ggatggacga aaagaccaca ggatggcgag gaggacacgt
    12301 ggtcgaggga ctggcaggag agctggaaca gctgcgggct agactggaac accatcctca
    12361 gggacagcga gagccaggaa gtggagcgat cgcgaaggaa gcctgcccca cagggctgta
    12421 cactcattct ggagaatgct gtaaagcttg taacctggga gagggagtgg cacagccatg
    12481 cggagccaat cagactgtgt gcgagccttg tctggactcc gtcacattct ctgatgtggt
    12541 cagtgccaca gaaccttgca agccatgtac tgagtgcgtg ggcctgcagt ctatgagtgc
    12601 tccttgtgtg gaggctgacg atgcagtctg ccggtgtgca tacggatact atcaggacga
    12661 gactaccggc agatgtgaag cttgcagggt gtgtgaggca ggctcagggc tggtctttag
    12721 ctgccaggat aaacagaaca ccgtgtgcga ggaatgtcct gacgggacat atagcgatga
    12781 ggccaatcac gtggacccct gcctgccttg tactgtgtgc gaggataccg aaaggcagct
    12841 gcgcgaatgt accagatggg cagacgccga gtgcgaggaa atcccagggc gatggattac
    12901 tcggtccacc ccccctgaag gatcagacag caccgcacca tctacacagg agccagaagc
    12961 accaccagag caggatctga tcgcctccac cgtggctggc gtggtcacaa ctgtcatggg
    13021 gagctcccag ccagtggtca cccggggcac cacagataac ctgattcccg tgtattgctc
    13081 catcctggca gccgtcgtcg tcggactggt ggcatacatc gccttcaagc ggcaattgat
    13141 gcacagcaga ggctctggcg agggcagagg aagtcttcta acatgcggtg acgtggagga
    13201 gaatcccggc ccttcaaacg ccaccatggt gtccaagggc gaagagaaca acatggccat
    13261 catcaaagaa ttcatgcggt tcaaggtgca catggaaggc agcgtgaacg gccacgagtt
    13321 cgagattgaa ggcgaaggcg agggcagacc ctacgaggct tttcagaccg ccaagctgaa
    13381 agtgaccaaa ggcggccctc tgccttttgc ctgggacatt ctgagccctc agtttatgta
    13441 cggcagcaag gtgtacatca agcaccccgc cgacattccc gactacttca agctgagctt
    13501 ccccgagggc ttcagatggg agagagtgat gaacttcgag gacggcggca tcatccacgt
    13561 gaaccaggat agctctctgc aggacggcgt gttcatctac aaagtgaagc tgcggggcac
    13621 caactttccc tctgatggcc ccgtgatgca gaaaaagacc atgggctggg aagccagcga
    13681 ggaaagaatg taccctgagg atggcgccct gaagtccgag atcaagaagc ggctgaagct
    13741 gaaggatggc ggccactatg ccgccgaagt gaaaaccacc tacaaggcca agaaacccgt
    13801 gcagctgcct ggcgcctaca tcgtggatat caagctggac atcgtgtccc acaacgagga
    13861 ctacaccatc gtggaacagt acgagagagc cgaaggcaga cacagcacag gcggaatgga
    13921 cgagctgtac aagtaaggta ccataacttc gtatagcata cattatacga agttataaac
    13981 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
    14041 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
    14101 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
    14161 cagcaa
//
