LOCUS       sequence-72207-e        9921 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human NKCC1 mutant lacking cysteine residues within 
            transmembrane domains and with an N-terminal 3xFLAG-YFP tag in 
            mammalian cells. cDNA is synthetic,containing convenient restriction
            sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9921)
  AUTHORS   Somasekharan S, Monette MY, Forbush B
  TITLE     Functional Expression of Human NKCC1 from a Synthetic Cassette-Based
            cDNA: Introduction of Extracellular Epitope Tags and Removal of 
            Cysteines.
  JOURNAL   PLoS One. 2013 Dec 5;8(12):e82060. doi: 
            10.1371/journal.pone.0082060.
  PUBMED    24339991
REFERENCE   2  (bases 1 to 9921)
  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: "PLoS One.";
            date: "2013-12-5"; volume: "8(12):e82060. doi"; pages: " 
            10.1371/journal.pone.0082060"
COMMENT     Sequence Label: pcDNA3.1 Flag YFP hNKCC1 TM-cysteineless (NT859)
FEATURES             Location/Qualifiers
     source          1..9921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      19..28
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             31..96
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(109..111,112..822)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(109..111,112..822)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(151..172)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(412..431)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     759..780
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4621..4638)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4627..4851
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4897..5325
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4984..5003)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5194..5215
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5334..5354)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5339..5668
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5519..5654
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5581..5600
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5735..6529
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5789..5808)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6399..6418
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6703..6824
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6740..6759)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6794..6813
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6873..6889)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6873..6889)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6886..6908)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6897..6913
                     /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(6921..6927,6928..6945,6946..6951))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6966..6987
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7104..7121)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7275..7860)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7355..7374)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8031..8822,8823..8891))
                     /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     8654..8673
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8892..8996)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9071..9090)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        9262..9641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9642..9845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9796..9816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9890..9909
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9890..9908
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gctagcccgg gcacggccgc caccatggga gactacaaag accatgacgg tgattataaa
       61 gatcacgaca tcgactacaa ggatgacgat gacaagggtg accctaccgt cagcaagggc
      121 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      181 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      241 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc
      301 ggctacggcc tgatgtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
      361 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
      421 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
      481 ctgaagggca tcgacttcaa ggaggacggc aacatcctag ggcacaagct ggagtacaac
      541 tacaacagcc acaacgttta tatcatggcc gacaagcaga agaacggcat caaggtgaac
      601 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
      661 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
      721 tccaaactga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
      781 accgccgccg ggatcactct cggcatggac gagctctaca agggtgacct ggagccgagg
      841 cccacggctc ctagctccgg cgcgccaggc ctcgccggag tcggcgaaac gccgagcgcc
      901 gctgcgctgg ccgcagccag ggtggaactg cccggaaccg ctgtgccctc ggtgccggag
      961 gatgctgcgc ccgcgagccg ggacgggggc ggggtccgcg atgagggacc agccgccgcc
     1021 ggtgacggcc tgggcagacc cttggggcct accccgagcc agagccgttt ccaggtggac
     1081 ctcgtttccg agaacgcagg gcgggctgct gctgcggcgg cggcggctgc tgcggccgcc
     1141 gccgctgctg gtgccggggc gggggccaag cagactcctg ccgacgggga agccagcggc
     1201 gagagcgagc cagctaaagg cagcgaggaa gccaaaggcc gcttccgcgt taacttcgta
     1261 gatcctgctg ccagctcgtc cgcagaagat agcctgtcag atgccgccgg ggtcggagtc
     1321 gatggaccca acgtgagctt ccagaacggc ggggacacgg tactgagcga gggcagcagc
     1381 ctccattccg gtggcggcgg cggctccggc caccaccagc actactatta tgatacccac
     1441 accaacacct actatctgcg caccttcggc cacaacacaa tggacgcggt acccaggatc
     1501 gaccactacc ggcacacagc cgcgcagctc ggcgagaagc tgctccggcc tagcctggcc
     1561 gagctgcatg acgagctgga aaaggaacct tttgaggatg gctttgcaaa tggggaagaa
     1621 agcactccaa cacgtgatgc tgtggtcacg tacacggccg aaagtaaagg agtcgtgaag
     1681 tttggctgga tcaagggtgt actggtacgt gccatgttaa acatttgggg tgtgatgctt
     1741 ttcattcgtc tctcatgcgt ctctgctgtg gtcacgtaca cggccgaaag taaaggagtc
     1801 gtgaagtttg gctggatcaa gggtgtacta gtacgttgta tgttaaacat ttggggtgtg
     1861 atgcttttca ttcgtctctc atggattgtg ggacaagctg gaattggtct atcagtcctt
     1921 gtaataatga tggctactgt tgtgacaact atcacaggat tgtccactag cgccatcgca
     1981 actaatggat ttgtaagggg aggcggcgca tattatttaa tatctagaag tctagggcct
     2041 gaatttggtg gtgccattgg tctaatcttc gcctttgcca acgctgttgc tgttgctatg
     2101 tatgtggttg gatttgcaga aaccgtggtg gagttgctga aagaacattc cattcttatg
     2161 atagatgaaa tcaatgacat ccggattatt ggagccatta cagtcgtgat tcttttaggt
     2221 atctcagtag ccggtatgga gtgggaagca aaagctcaga ttgttctttt ggtgatccta
     2281 cttcttgcta ttggtgattt cgtcatagga acatttatcc cactcgagag caagaagcca
     2341 aaagggtttt ttggctataa atctgaaatc tttaatgaga actttggccc cgattttcga
     2401 gaagaagaaa ctttcttttc tgtatttgcc atcttttttc ctgctgcaac tggaattctg
     2461 gctggcgcaa atatctcagg tgatcttgca gatcctcagt cagccatacc caaaggaaca
     2521 ctcctagcca ttttaattac tacattggtt tatgtaggaa ttgccgtgtc tgtaggttct
     2581 atggtcgtac gcgatgccac tggtaatgtt aatgacacta tcgtaacaga gctaacaaac
     2641 tgtactagcg cagcctgcaa attaaacttt gatttttcat cttgtgaatc cagtccttgt
     2701 agctacggcc taatgaacaa ctttcaggta atgagtatgg tgtcaggatt tacaccacta
     2761 atttccgcgg gcatattctc tgccactctg tcttccgcat tagcatccct agtgagtgct
     2821 cccaagatat ttcaggccct atccaaagac aacatctacc ccgctttcca gatgtttgct
     2881 aaaggttatg ggaaaaataa tgagccatta cgtggctaca tcttaacatt cttaattgca
     2941 cttggattca tcttaattgc cgaactgaat gttattgcac caattatctc aaacttcttc
     3001 cttgccagct atgccctcat caatttttca gtattccatg cttcactggc aaaaagccca
     3061 ggatggcgac cggcatttaa atactacaat atgtggatct cacttcttgg agcaattctt
     3121 tccgtgatag taatgttcgt cattaactgg tgggctgctt tgctaaccta cgtgatagtc
     3181 cttggtctct atatttatgt tacctacaaa aaacccgatg tgaattgggg atcctctaca
     3241 caagccctga cttacctcaa tgccctgcaa catagtatta ggctttctgg cgtggaggac
     3301 catgtgaaaa actttaggcc acagtgtctt gttatgacag gagcgccaaa ctcacggccg
     3361 gccttacttc atcttgttca cgatttcaca aaaaatgttg gtttgatgat ctgtggacac
     3421 gtacacatgg gaccacgcag acaagccatg aaagaaatgt ccatcgacca agccaaatat
     3481 cagcgctggc ttattaagaa caaaatgaag gctttttatg caccagtaca tgcagatgac
     3541 ctcagagaag gggcccaata cttgatgcag gctgctggcc tgggccgtat gaagccaaac
     3601 acactagtcc ttggatttaa gaaagattgg ttgcaagcag atatgaggga tgtggatatg
     3661 tacataaact tatttcacga tgcttttgac atacaatatg gcgtagtggt tattcgccta
     3721 aaagaaggtc tggatattag tcacctgcag ggacaagaag aattattgtc atcacaagag
     3781 aaatctcctg gcaccaagga tgtcgtagta agtgtggaat atagtaaaaa gtccgattta
     3841 gatacttcga aacccctcag tgaaaaacca attacccata aagtcgagga agaagatggc
     3901 aagactgcaa ctcaaccact gttgaaaaaa gaatccaaag gccctattgt gcctttgaat
     3961 gtagctgacc aaaagcttct tgaagctagt acacagtttc agaaaaaaca aggaaagaat
     4021 actattgatg tctggtggct ttttgatgac ggaggtctga ccttattgat accttacctt
     4081 ctgacgacca agaaaaaatg gaaagactgt aagatcagag tattcattgg tggaaagata
     4141 aacagaatag accatgaccg gagagcgatg gctactttac tgagcaagtt cagaatcgat
     4201 ttttctgaca tcatggttct aggagacatc aataccaaac caaagaaaga aaacattata
     4261 gctttcgagg aaatcattga gccatacaga cttcacgaag atgataaaga gcaagatatc
     4321 gcagataaaa tgaaagaaga tgaaccctgg cgaataacag ataatgaatt ggagctgtat
     4381 aagaccaaga cataccggca gatcaggtta aatgagttat taaaggaaca ttcaagcaca
     4441 gctaacatta ttgtgatgag tttaccggtc gcacgaaagg gtgccgtttc tagtgctctc
     4501 tacatggcat ggttagaagc actatctaag gatctaccac caatcctact agttcgtggg
     4561 aatcatcaga gtgtccttac cttctattca taattaatta agtttaaacc cgctgatcag
     4621 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     4681 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     4741 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     4801 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     4861 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     4921 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4981 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5041 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5101 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5161 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5221 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     5281 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     5341 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5401 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     5461 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     5521 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5581 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     5641 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     5701 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     5761 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     5821 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     5881 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     5941 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     6001 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     6061 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     6121 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     6181 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     6241 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     6301 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     6361 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     6421 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     6481 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     6541 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     6601 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     6661 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     6721 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6781 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     6841 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6901 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6961 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     7021 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     7081 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     7141 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     7201 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7261 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7321 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7381 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7441 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     7501 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7561 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7621 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7681 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     7741 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7801 gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7861 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7921 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7981 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     8041 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     8101 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     8161 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     8221 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     8281 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     8341 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     8401 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8461 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8521 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8581 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8641 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8701 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8761 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8821 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8881 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8941 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     9001 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     9061 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     9121 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     9181 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     9241 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
     9301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     9361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     9421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     9481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     9541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     9601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     9661 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     9721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     9781 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     9841 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
     9901 actataggga gacccaagct g
//
