LOCUS       sequence-449962-        8895 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Nuclease active S. parasanguinis Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8895)
  AUTHORS   Butterfield GL, Rohm D, Roberts A, Nethery MA, Rizzo AJ, Morone DJ, 
            Garnier L, Iglesias N, Barrangou R, Gersbach CA
  TITLE     Characterization of diverse Cas9 orthologs for genome and epigenome 
            editing.
  JOURNAL   Proc Natl Acad Sci U S A. 2025 Mar 18;122(11):e2417674122. doi: 
            10.1073/pnas.2417674122. Epub 2025 Mar 12.
  PUBMED    40073054
REFERENCE   2  (bases 1 to 8895)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2025-03-18"; volume: "122(11):e2417674122. 
            doi"; pages: " 10.1073/pnas.2417674122. Epub 2025 Mar 12"
COMMENT     Sequence Label: pcDNA-Spa-Cas9
FEATURES             Location/Qualifiers
     source          1..8895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             907..972
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             982..1002
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4444..4464
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4486..4503)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4492..4716
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4762..5190
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4849..4868)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5059..5080
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5199..5219)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5204..5533
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5384..5519
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5446..5465
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5600..6394
                     /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(5654..5673)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6264..6283
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6568..6689
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6605..6624)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6659..6678
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6738..6754)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6738..6754)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6751..6773)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6762..6778
                     /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(6786..6792,6793..6810,6811..6816))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6831..6852
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6969..6986)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7140..7728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7220..7239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7899..8690,8691..8759))
                     /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     8522..8541
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8760..8864)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gccatggact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
      961 gacgatgaca agcacgttaa ccccaagaag aagaggaagg tgggccgcgg aaccggtatg
     1021 aacggactgg ttttgggtct tgatatcggg atcgctagcg ttggggtggg catcctgaag
     1081 aaggacatag gagaaatcat tcataccaat tctagactgt tttcagctgc cacagccgac
     1141 tctaatattg aacgacgagg acatcgtggc ggcaagaggc tgacaagacg aaaaaaacac
     1201 cgaagcatac gacttcacga tcttttcgag gattttggac tgctgacgga cttttcaaaa
     1261 gtttccatca acttgaatcc gtaccagtta cgcgtacagg gtctggacaa ccagctgaca
     1321 aacgaggagc tgtttatcgc tcttaagaat atcgtgaagc gccgggggat tagctactta
     1381 gacgatgcct ctgaagacgg cggaaccgtg tcttctgatt atgggaaggc tgtcgaagaa
     1441 aatagaaaac tcttagccga acagactcct gggcagatcc agctggacag attcgaaaag
     1501 tacggccaag tccgaggcga cttcaatgtc gtggagaacg gtgagaaacg acgtctgatt
     1561 aacgtcttta caactagcgc ctattccaag gaggccgaga ggatactgag gaagcagcag
     1621 gagttcaaca agaaaataac ggatgagttc atcgaggatt acctgaccat tcttactgga
     1681 aaaagaaaat attaccatgg tcctggaaac gaaaagtccc ggaccgatta cgggcggtac
     1741 acaaccaaaa aggacccaga gggcaaatac atcaccctcg ataatatttt cgggatcctc
     1801 atcggtaaat gcacttttta ccccgatgag tatcgcgcgt ctaaagcttc atataccgca
     1861 caggagttca atctgctcaa cgacctgaat aacctgaccg tgcctaccga aaccaaaaaa
     1921 ctgtcagagg agcagaagaa gacgataata aaatacgcca aaacggctaa gacccttggc
     1981 gcttctactc tgctgaagta tatagccaaa ctgatcggtg cttccgttga ccagattcac
     2041 gggtatagaa tcgacccaaa taaaaagccc gaaatgcaca ccttcgagac gtaccggaaa
     2101 atgcaatccc tggagacgat ctcagtggag gaactgcctc gcaaagtgct tgacgaactc
     2161 gcccatattc tgacattgaa cactgagcgc gaaggcatcg aggaggctat taatgccacc
     2221 ttgaaagata cgtttagtca ggaccaggtc ctcgaactcg tgcagttccg caaaaataac
     2281 tcttccttat tctcaaaggg atggcatagt ttcagcctga aactgatgat ggaactgatt
     2341 cctgaactct atgagactag tgaagagcag atgactatac tgactcgtct ggggaaacag
     2401 aagtctaagg agacaagtaa acgaactaag tacattgatg aaaaggagct gacagaagag
     2461 atttataatc cagtcgtggc taaatccgtc cgtcaggcta ttaagatcat taacgaggca
     2521 acgaaaaagt acggaatctt cgataacatt gtgatcgaaa tggcccgtga gaacaatgaa
     2581 gaagatgcta agaaggagta catcaagcgg cagaaggcaa acttggatga gaagaacgcc
     2641 gcaatggaga aagctgcttt tcaatacaac ggtaagaagg aactcccgga taacgtcttc
     2701 cacggccata aggagctcgc cacaaaaata cggttgtggc accagcaggg ggaaaagtgc
     2761 ctctacactg gaaaaaatat ccctatctcc gaccttattc aaaatcagta caagtatgaa
     2821 atcgaccaca tcctcccact gtccctcagt ttcgacgata gcctgtccaa caaggtcctc
     2881 gtgctggcta ccgccaatca ggaaaagggc caaagaactc cttttcaagc tctcgattca
     2941 atggatgacg cctggagcta tcgggagttc aaatcctatg tgaaggattc taaactcttg
     3001 gggaataaaa agaaggaata cttgttaaca gaggaggata tcagtaagat cgaggtgaaa
     3061 cagaagttta tcgaacggaa tttagttgac acaaggtact caagtcgcgt ggttcttaac
     3121 gcccttcagg acttctacaa ggaacaccag ttcgatacca ccattagcgt ggttagagga
     3181 caatttacat cccagctgcg gcggaaatgg ggactggaga agagccgaga aacctaccat
     3241 caccacgccg tggacgcgct gattattgcc gccagtagcc agctccgcct ctggaaaaaa
     3301 cagaacaatc ctcttatcag ttacaccgaa ggccagtttg tggatcaggt gaccggcgag
     3361 attatatccc tttctgacga cgaatacaaa gagctggttt tcaaagctcc atatgaccac
     3421 tttgttgaca cactgaagtc aaagaagttt gaggattcta tcctcttttc ataccaggtg
     3481 gattctaagt acaaccggaa gattagcgac gcaaccatat atgcaactcg taaagccaaa
     3541 ttggacaagg agaataaaga gtatacttac actctgggta aaatcaaaga tatctatgct
     3601 cttggcacga aaagcccttc taaaaccggc ttttacaagt ttctggacct gtataataaa
     3661 gacaagtctc agttcctgat gtttcagaaa gatcgcaaga cctgggacga ggtgattgaa
     3721 aagatcattg agcaataccg gccttttaaa gaatacgacg aaaacggaaa agaagtagac
     3781 ttcaaccctt ttgaaaagta ccggatcgaa aatggcccca tccgaaaata ttccaaaaag
     3841 ggcaatggcc cagaaataaa gagtctcaag tattacgata acttactggg taagtttgtg
     3901 gatatcacac cttcagagtc caagaacccc gtagctctgc tgtccctgaa cccgtggagg
     3961 actgacgtct actacaacac cgaaacatcc aagtatgagt tccttgggct gaagtatgct
     4021 gatttgtgct tcgaaaaggg gggtgcatat ggtatctctg aggtgaaata caataaaata
     4081 agagagaaag aaggtatcgg aaaggagtct gaattcaaat ttacccttta taagaatgac
     4141 ctgattctga tcaaagatac agagacaaac tgccaacaga tttttcgctt ctggtctcga
     4201 actggaaaag acaatcctaa atcatttgag aagcataaaa ttgagcttaa accgtatgaa
     4261 aaggctcgtt tcgagaaggg tgaagaactg gaggtgctgg gaaaggtgcc acctagttca
     4321 aaccaactgc agaaaaatat gcaaattgag aacctctcca tctacaaggt gaagactgat
     4381 gtgcttggca ataagcattt tatcaaaaag gaaggagaag aacctaagct caaatttacc
     4441 ggtcctaaga aaaagcggaa agtggctgtt taaaccgctg atcagcctcg actgtgcctt
     4501 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     4561 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     4621 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     4681 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     4741 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4801 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4861 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4921 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4981 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5041 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5101 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5161 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     5221 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     5281 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5341 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     5401 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     5461 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     5521 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     5581 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     5641 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     5701 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     5761 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     5821 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     5881 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5941 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     6001 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     6061 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     6121 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     6181 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     6241 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     6301 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     6361 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     6421 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     6481 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     6541 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     6601 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6661 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     6721 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     6781 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6841 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6901 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6961 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     7021 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     7081 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     7141 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     7201 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     7261 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7321 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7381 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7441 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7501 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7561 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7621 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7681 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7741 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7801 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7861 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7921 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7981 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     8041 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     8101 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     8161 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     8221 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     8281 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     8341 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     8401 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8461 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8521 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8581 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8641 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8701 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8761 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8821 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8881 gtgccacctg acgtc
//
