LOCUS       sequence-396626-       11451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses RFP and four sgRNAs against GBX2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11451)
  AUTHORS   Maimaitili M, Chen M, Febbraro F, Ucuncu E, Kelly R, Niclis JC, 
            Christiansen JR, Mermet-Joret N, Niculescu D, Lauritsen J, Iannielli
            A, Klaestrup IH, Jensen UB, Qvist P, Nabavi S, Broccoli V, Nykjaer 
            A, Romero-Ramos M, Denham M
  TITLE     Enhanced production of mesencephalic dopaminergic neurons from 
            lineage-restricted human undifferentiated stem cells.
  JOURNAL   Nat Commun. 2023 Dec 5;14(1):7871. doi: 10.1038/s41467-023-43471-0.
  PUBMED    38052784
REFERENCE   2  (bases 1 to 11451)
  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: "Nat 
            Commun."; date: "2023-12-5"; volume: "14(1):7871. doi"; pages: " 
            10.1038/s41467-023-43471-0"
COMMENT     Sequence Label: pLV-4gRNA-GBX2-RFP
FEATURES             Location/Qualifiers
     source          1..11451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        33..1244
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1081..1100
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      1291..1300
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1297..1974
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(1477..1497)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1695..1714
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1889..1912
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    1999..2587
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2052..2072)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2470..2481)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2590..2606)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2591..2607)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3112..3292
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(3318..3335)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3324..3548
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3594..4022
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3681..3700)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3891..3912
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4031..4051)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4036..4365
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4216..4351
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4278..4297
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4413..4460
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4479..4853
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4983..5104
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5020..5039)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5074..5093
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5153..5169)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5153..5169)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5166..5188)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5177..5193
                     /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(5201..5207,5208..5225,5226..5231))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5246..5267
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5384..5401)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5555..6143)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5635..5654)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6314..7105,7106..7174))
                     /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     6937..6956
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7175..7279)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7354..7373)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7545..7924
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7926..8124
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8079..8099
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             8142..8322
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8369..8494
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8987..9220
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9405..9449
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    9747..9864
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(9909..9942)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        9947..10260
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     10179..10199
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     misc_RNA        10283..10358
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(10388..10463)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(10485..10733)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(10543..10562)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(10713..10733)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        10763..10977
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     10898..10917
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_RNA        11007..11082
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(11112..11187)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 acaattaatt aacccgtgtc ggctccagat ctggcctccg cgccgggttt tggcgcctcc
       61 cgcgggcgcc cccctcctca cggcgagcgc tgccacgtca gacgaagggc gcagcgagcg
      121 tcctgatcct tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt
      181 agaaccccag tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact
      241 ggttttcttt ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc
      301 ggagggatct ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc
      361 acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga
      421 tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc cgccgggccg
      481 ctcggtggga cggaagcgtg tggagagacc gccaagggct gtagtctggg tccgcgagca
      541 aggttgccct gaactggggg ttggggggag cgcagcaaaa tggcggctgt tcccgagtct
      601 tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg
      661 tgggcggcaa gaacccaagg tcttgaggcc ttcgctaatg cgggaaagct cttattcggg
      721 tgagatgggc tggggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa
      781 ctcggtttgt cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc
      841 gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata
      901 atgcagggtg gggccacctg ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc
      961 agggttcggg cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg
     1021 agggataagt gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc
     1081 tgaagctccg gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt
     1141 aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagactagta
     1201 aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacgaagct tgggctgcag
     1261 gtcgactcta gaggatcccc gggtaccggt gccaccatgg atagcactga gaacgtcatc
     1321 aagcccttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
     1381 atcgagggcg agggcgaggg caagccctac gagggcaccc agaccgccaa gctgcaggtg
     1441 accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc
     1501 tccaaggtgt acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc
     1561 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     1621 caggactcct ccctgcagga cggcaccttc atctaccacg tgaagttcat cggcgtgaac
     1681 ttcccctccg acggccccgt aatgcagaag aagactctgg gctgggagcc ctccaccgag
     1741 cgcctgtacc cccgcgacgg cgtgctgaag ggcgagatcc acaaggcgct gaagctgaag
     1801 ggcggcggcc actacctggt ggagttcaag tcaatctaca tggccaagaa gcccgtgaag
     1861 ctgcccggct actactacgt ggactccaag ctggacatca cctcccacaa cgaggactac
     1921 accgtggtgg agcagtacga gcgcgccgag gcccgccacc acctgttcca gtaggaattc
     1981 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2041 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2101 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     2161 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     2221 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     2281 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2341 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     2401 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     2461 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     2521 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     2581 tccccgcatc gataccgtcg acctcgagac ctagaaaaac atggagcaat cacaagtagc
     2641 aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg
     2701 ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat
     2761 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     2821 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaac
     2881 tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta caagctagta
     2941 ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg cttgttacac
     3001 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg gaggtttgac
     3061 agccgcctag catttcatca catggcccga gagctgcatc cggactgtac tgggtctctc
     3121 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     3181 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     3241 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagggcccgt
     3301 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3361 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3421 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3481 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3541 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3601 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3661 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3721 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3781 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3841 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3901 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3961 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4021 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4081 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4141 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4201 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4261 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4321 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4381 tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata
     4441 tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg accagtgccg
     4501 ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg
     4561 ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc
     4621 tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg
     4681 tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg
     4741 acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc
     4801 tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc
     4861 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4921 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4981 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5041 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5101 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5161 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5221 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5281 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5341 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5401 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5461 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5521 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5581 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5641 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5701 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5761 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5821 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5881 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5941 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6001 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6061 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6121 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6181 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6241 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6301 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6361 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6421 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6481 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6541 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6601 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6661 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6721 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6781 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6841 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6901 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6961 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7021 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7081 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7141 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7201 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7261 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     7321 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     7381 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     7441 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     7501 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
     7561 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     7621 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     7681 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     7741 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     7801 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     7861 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     7921 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     7981 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     8041 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     8101 acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc
     8161 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     8221 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     8281 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     8341 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
     8401 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
     8461 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     8521 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     8581 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
     8641 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     8701 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     8761 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
     8821 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     8881 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     8941 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     9001 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     9061 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     9121 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     9181 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     9241 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     9301 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     9361 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     9421 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     9481 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     9541 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     9601 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
     9661 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc
     9721 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     9781 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     9841 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg
     9901 ttaattaaat aacttcgtat agcatacatt atacgaagtt atgatagatc cgacgccgcc
     9961 atctctaggc ccgcgccggc cccctcgcac agacttgtgg gagaagctcg gctactcccc
    10021 tgccccggtt aatttgcata taatatttcc tagtaactat agaggcttaa tgtgcgataa
    10081 aagacagata atctgttctt tttaatacta gctacatttt acatgatagg cttggatttc
    10141 tataagagat acaaatacta aattattatt ttaaaaaaca gcacaaaagg aaactcaccc
    10201 taactgtaaa gtaattgtgt gttttgagac tataaatatc ccttggagaa aagccttgtt
    10261 tgggcagcac taccggccgg tagttttaga gctagaaata gcaagttaaa ataaggctag
    10321 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttgtacg ctcagcggag
    10381 aaaaaaagca ccgactcggt gccacttttt caagttgata acggactagc cttattttaa
    10441 cttgctattt ctagctctaa aacagcgggc gctgcatcat catcggtgtt tcgtcctttc
    10501 cacaagatat ataaagccaa gaaatcgaaa tactttcaag ttacggtaag catatgatag
    10561 tccattttaa aacataattt taaaactgca aactacccaa gaaattatta ctttctacgt
    10621 cacgtatttt gtactaatat ctttgtgttt acagtcaaat taattccaat tatctctcta
    10681 acagccttgt atcgtatatg caaatatgaa ggaatcatgg gaaataggcc ctcttcctgc
    10741 ccgaccttgg atcgatccgc ctgaacgctg acgtcatcaa cccgctccaa ggaatcgcgg
    10801 gcccagtgtc actaggcggg aacacccagc gcgcgtgcgc cctggcagga agatggctgt
    10861 gagggacagg ggagtggcgc cctgcaatat ttgcatgtcg ctatgtgttc tgggaaatca
    10921 ccataaacgt gaaatgtctt tggatttggg aatcttataa gttctgtatg agaccactct
    10981 ttcccaaaag gtggaagacg acccgagttt tagagctaga aatagcaagt taaaataagg
    11041 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttg gtcatcagtc
    11101 cgagaaaaaa agcaccgact cggtgccact ttttcaagtt gataacggac tagccttatt
    11161 ttaacttgct atttctagct ctaaaacgcc gtcgctgatg atgatgccga ggtacccaag
    11221 cggcgcacaa gctatataaa cctgaaggaa gtctcaactt tacacttagg tcaagttgct
    11281 tatcgtacta gagcttcagc aggaaattta actaaaatct aatttaacca gcatagcaaa
    11341 tatcatttat tcccaaaatg ctaaagtttg agataaacgg acttgatttc cggctgtttt
    11401 gacactatcc agaatgcctt gcagatgggt ggggcatgct aaatactgca g
//
