LOCUS       sequence-76355-e        9795 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human NKCC1 F728C mutant 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 9795)
  TITLE     Forbush Lab human NKCC1 NT clones
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9795)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pcDNA3.1 Flag YFP hNKCC1 F728C (NT502)
FEATURES             Location/Qualifiers
     source          1..9795
                     /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(4495..4512)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4501..4725
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4771..5199
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4858..4877)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5068..5089
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5208..5228)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5213..5542
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5393..5528
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5455..5474
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5609..6403
                     /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(5663..5682)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6273..6292
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6577..6698
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6614..6633)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6668..6687
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6747..6763)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6747..6763)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6760..6782)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6771..6787
                     /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(6795..6801,6802..6819,6820..6825))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6840..6861
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6978..6995)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7149..7734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7229..7248)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7905..8696,8697..8765))
                     /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     8528..8547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8766..8870)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8945..8964)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        9136..9515
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9516..9719
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9670..9690
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9764..9783
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9764..9782
                     /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 actagtacgt tgtatgttaa acatttgggg tgtgatgctt
     1741 ttcattcgtc tctcatggat tgtgggacaa gctggaattg gtctatcagt ccttgtaata
     1801 atgatggcta ctgttgtgac aactatcaca ggattgtcca ctagcgccat cgcaactaat
     1861 ggatttgtaa ggggaggcgg cgcatattat ttaatatcta gaagtctagg gcctgaattt
     1921 ggtggtgcca ttggtctaat cttcgccttt gccaacgctg ttgctgttgc tatgtatgtg
     1981 gttggatttg cagaaaccgt ggtggagttg ctgaaagaac attccattct tatgatagat
     2041 gaaatcaatg acatccggat tattggagcc attacagtcg tgattctttt aggtatctca
     2101 gtagccggta tggagtggga agcaaaagct cagattgttc ttttggtgat cctacttctt
     2161 gctattggtg atttcgtcat aggaacattt atcccactcg agagcaagaa gccaaaaggg
     2221 ttttttggct ataaatctga aatctttaat gagaactttg gccccgattt tcgagaagaa
     2281 gaaactttct tttctgtatt tgccatcttt tttcctgctg caactggaat tctggctggc
     2341 gcaaatatct caggtgatct tgcagatcct cagtcagcca tacccaaagg aacactccta
     2401 gccattttaa ttactacatt ggtttatgta ggaattgccg tgtctgtagg ttcttgtgtc
     2461 gtacgcgatg ccactggtaa tgttaatgac actatcgtaa cagagctaac aaactgtact
     2521 agcgcagcct gcaaattaaa ctttgatttt tcatcttgtg aatccagtcc ttgtagctac
     2581 ggcctaatga acaactttca ggtaatgagt atggtgtcag gatttacacc actaatttcc
     2641 gcgggcatat tctctgccac tctttctagc gcattagcat ccctagtgag tgctcccaag
     2701 atatttcagg ccctatgtaa agacaacatc taccccgctt tccagatgtt tgctaaaggt
     2761 tatgggaaaa ataatgagcc attacgtggc tacatcttaa cattcttaat tgcacttgga
     2821 ttcatcttaa ttgccgaact gaatgttatt gcaccaatta tctcaaactt cttccttgcc
     2881 agctatgccc tcatcaattt ttcagtattc catgcttcac tggcaaaaag cccaggatgg
     2941 cgaccggcat ttaaatacta caatatgtgg atctcacttc ttggagcaat tctttgttgc
     3001 atagtaatgt gtgtcattaa ctggtgggct gctttgctaa cctacgtgat agtccttggt
     3061 ctctatattt atgttaccta caaaaaaccc gatgtgaatt ggggatcctc tacacaagcc
     3121 ctgacttacc tcaatgccct gcaacatagt attaggcttt ctggcgtgga ggaccatgtg
     3181 aaaaacttta ggccacagtg tcttgttatg acaggagcgc caaactcacg gccggcctta
     3241 cttcatcttg ttcacgattt cacaaaaaat gttggtttga tgatctgtgg acacgtacac
     3301 atgggaccac gcagacaagc catgaaagaa atgtccatcg accaagccaa atatcagcgc
     3361 tggcttatta agaacaaaat gaaggctttt tatgcaccag tacatgcaga tgacctcaga
     3421 gaaggggccc aatacttgat gcaggctgct ggcctgggcc gtatgaagcc aaacacacta
     3481 gtccttggat ttaagaaaga ttggttgcaa gcagatatga gggatgtgga tatgtacata
     3541 aacttatttc acgatgcttt tgacatacaa tatggcgtag tggttattcg cctaaaagaa
     3601 ggtctggata ttagtcacct gcagggacaa gaagaattat tgtcatcaca agagaaatct
     3661 cctggcacca aggatgtcgt agtaagtgtg gaatatagta aaaagtccga tttagatact
     3721 tcgaaacccc tcagtgaaaa accaattacc cataaagtcg aggaagaaga tggcaagact
     3781 gcaactcaac cactgttgaa aaaagaatcc aaaggcccta ttgtgccttt gaatgtagct
     3841 gaccaaaagc ttcttgaagc tagtacacag tttcagaaaa aacaaggaaa gaatactatt
     3901 gatgtctggt ggctttttga tgacggaggt ctgaccttat tgatacctta ccttctgacg
     3961 accaagaaaa aatggaaaga ctgtaagatc agagtattca ttggtggaaa gataaacaga
     4021 atagaccatg accggagagc gatggctact ttactgagca agttcagaat cgatttttct
     4081 gacatcatgg ttctaggaga catcaatacc aaaccaaaga aagaaaacat tatagctttc
     4141 gaggaaatca ttgagccata cagacttcac gaagatgata aagagcaaga tatcgcagat
     4201 aaaatgaaag aagatgaacc ctggcgaata acagataatg aattggagct gtataagacc
     4261 aagacatacc ggcagatcag gttaaatgag ttattaaagg aacattcaag cacagctaac
     4321 attattgtga tgagtttacc ggtcgcacga aagggtgccg tttctagtgc tctctacatg
     4381 gcatggttag aagcactatc taaggatcta ccaccaatcc tactagttcg tgggaatcat
     4441 cagagtgtcc ttaccttcta ttcataatta attaagttta aacccgctga tcagcctcga
     4501 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     4561 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     4621 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     4681 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     4741 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     4801 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4861 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4921 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4981 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5041 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5101 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     5161 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     5221 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5281 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     5341 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     5401 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     5461 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     5521 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     5581 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     5641 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     5701 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     5761 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     5821 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     5881 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     5941 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     6001 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     6061 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     6121 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     6181 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     6241 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     6301 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     6361 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     6421 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     6481 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     6541 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     6601 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6661 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     6721 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     6781 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     6841 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6901 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6961 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     7021 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     7081 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     7141 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7201 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7261 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7321 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     7381 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7441 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7501 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7561 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     7621 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7681 agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     7741 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     7801 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     7861 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     7921 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     7981 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     8041 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     8101 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     8161 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     8221 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     8281 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     8341 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     8401 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     8461 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     8521 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     8581 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     8641 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     8701 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     8761 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     8821 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     8881 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
     8941 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     9001 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     9061 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
     9121 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
     9181 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     9241 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     9301 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     9361 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     9421 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     9481 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     9541 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     9601 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     9661 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     9721 tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg actcactata
     9781 gggagaccca agctg
//
