LOCUS       sequence-483779-        7577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the SIBR-Int4-Cas9 system and a NT sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7577)
  AUTHORS   Della Valle S, Orsi E, Creutzburg SCA, Jansen LFM, Pentari E, Beisel
            CL, Steel H, Nikel PI, Staals RHJ, Claassens NJ, van der Oost J, 
            Huang WE, Patinios C
  TITLE     Streamlined and efficient genome editing in Cupriavidus necator H16 
            using an optimised SIBR-Cas system
  JOURNAL   bioRxiv 2024.11.24.625072; doi: 10.1101/2024.11.24.625072
REFERENCE   2  (bases 1 to 7577)
  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: "bioRxiv 
            2024.11.24.625072; doi: 10.1101/2024.11.24.625072"
COMMENT     Sequence Label: pSIBR-Int4-Cas9-NT
FEATURES             Location/Qualifiers
     source          1..7577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4102..4119
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        join(4198..4203,4204..4221,4222..4228)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     misc_RNA        4264..4339
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(4613..4632)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4624..5439
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5510..6172)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      6173..6944
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      complement(7059..7145)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     7160..7182
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     promoter        join(7189..7194,7195..7212,7213..7219)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             join(7572..7577,1..4101)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MLDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIK
                     KNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEE
                     SFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMI
                     KFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRR
                     LENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLA
                     QIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALV
                     RQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDL
                     LRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLAR
                     GNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYE
                     YFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECF
                     DSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEE
                     RLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNF
                     MQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMG
                     RHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKL
                     YLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV
                     PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITK
                     HVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAY
                     LNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFK
                     TEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFS
                     KESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLG
                     ITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGN
                     ELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILA
                     DANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEV
                     LDATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(7572..7577,1..4101)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MLDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIK
                     KNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEE
                     SFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMI
                     KFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRR
                     LENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLA
                     QIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALV
                     RQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDL
                     LRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLAR
                     GNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYE
                     YFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECF
                     DSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEE
                     RLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNF
                     MQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMG
                     RHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKL
                     YLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV
                     PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITK
                     HVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAY
                     LNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFK
                     TEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFS
                     KESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLG
                     ITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGN
                     ELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILA
                     DANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEV
                     LDATLIHQSITGLYETRIDLSQLGGD"
ORIGIN
        1 gacaaaaagt actcgattgg gctcgacatc gggaccaact ccgttgggtg ggctgttatc
       61 accgacgagt acaaagtccc ttcgaagaaa tttaaagtct tggggaacac cgatcgtcat
      121 agcatcaaga agaaccttat tggagcgctt ctcttcgata gcggggaaac cgctgaggct
      181 acccggttga agcgaaccgc gcggaggagg tacacccggc gaaaaaaccg gatctgctac
      241 cttcaggaaa tcttctcgaa cgaaatggct aaggttgacg acagcttttt ccaccgtctt
      301 gaggaatcgt tccttgttga ggaggataaa aaacacgagc ggcaccccat cttcgggaac
      361 attgttgacg aggtcgcgta ccacgaaaag taccccacca tctaccactt gcgtaagaag
      421 cttgttgact cgaccgacaa ggctgacctt cgtctcatct accttgcact cgctcacatg
      481 atcaaattcc gggggcactt ccttatcgaa ggggacctca accccgacaa cagcgacgtt
      541 gataagcttt tcatccagct tgttcagacg tacaaccagc tcttcgagga gaaccccatc
      601 aacgctagcg gggttgacgc gaaggctatc ctttcggctc gtcttagcaa gtcgaggcgt
      661 ctcgagaact tgatcgcgca gttgcctggg gaaaaaaaga acgggctctt cggaaacttg
      721 atcgcgcttt cgcttgggct tacgcccaac ttcaagtcga acttcgacct tgctgaggac
      781 gcgaagctcc agctttcgaa ggacacctac gacgacgacc tcgacaacct ccttgctcag
      841 atcggggacc agtacgcgga ccttttcctt gctgcgaaaa acctctcgga cgcgatcctc
      901 ctttcggaca tccttagggt taacaccgag attaccaaag cgcctctttc ggcgtcgatg
      961 atcaagcgtt acgacgagca ccaccaggat cttacccttc tcaaggcgct cgtccgtcag
     1021 cagcttcccg agaaatacaa ggagatcttc ttcgaccagt cgaagaacgg gtacgctggg
     1081 tacatcgacg gaggggcgtc ccaggaggag ttctacaagt tcatcaagcc catcctcgag
     1141 aagatggacg ggaccgaaga gctccttgtt aagcttaacc gggaggacct tttgcgtaaa
     1201 cagcgaacgt tcgataacgg gtcgatccct caccagatcc atcttgggga attgcacgcg
     1261 atccttagga ggcaggagga tttctacccc ttcctcaagg ataaccggga aaaaatcgag
     1321 aagatcctta ccttccgtat cccctactac gtcgggcccc ttgctcgggg gaacagccgg
     1381 ttcgcttgga tgacccgaaa atcggaggag accatcacgc cctggaactt cgaggaggtc
     1441 gttgacaagg gggcgtcggc gcagtcgttc atcgagcgta tgaccaactt cgacaagaac
     1501 cttcccaacg agaaggttct tcccaagcac agccttcttt acgaatactt cacggtctac
     1561 aacgagctta ccaaagttaa gtacgtcacc gaggggatgc gtaagcccgc tttcctttcg
     1621 ggggagcaga aaaaggcaat cgtcgacctc ttgtttaaga ccaaccgtaa ggttacggtc
     1681 aaacagctca aggaggacta ctttaagaag attgagtgct tcgacagcgt cgagatctcg
     1741 ggggtcgagg acaggttcaa cgcgtcgctc gggacgtacc acgaccttct taagatcatc
     1801 aaggacaagg acttccttga caacgaggag aacgaggaca tcctcgaaga catcgtcctc
     1861 acccttacgc tcttcgagga cagggaaatg atcgaagaga ggcttaagac ctacgcgcat
     1921 ttgttcgacg acaaagttat gaagcagctt aagcggcgtc ggtacaccgg gtgggggcgg
     1981 ctttcgcgta agcttatcaa cgggatcagg gacaaacagt cggggaagac cattctcgac
     2041 ttccttaagt cggacgggtt cgcaaaccgt aacttcatgc agttgatcca cgacgacagc
     2101 cttaccttca aggaggatat ccagaaggct caggtttcgg ggcaggggga cagcctccac
     2161 gagcacatcg ctaacctcgc ggggtccccc gcgatcaaga aggggatcct ccagaccgtt
     2221 aaggtcgtcg acgagcttgt taaggttatg ggacgacaca aacccgagaa catcgtcatc
     2281 gagatggctc gggagaacca gaccacccag aaagggcaga agaactcccg tgaacggatg
     2341 aagcgtatcg aggaggggat caaggagctc gggagccaga tccttaagga acaccccgtc
     2401 gagaacaccc agcttcagaa cgagaaattg tacttgtact acttgcagaa cgggagggat
     2461 atgtacgttg atcaggagct cgacatcaac cggctcagcg actacgacgt tgaccatatc
     2521 gtcccccaga gctttcttaa ggatgactcg attgataaca aagttctcac gcggtcggac
     2581 aagaaccggg ggaagtccga caacgtcccc agcgaggagg ttgttaagaa aatgaagaac
     2641 tactggaggc agcttcttaa cgcaaaactc atcacccagc ggaaattcga caacctcacg
     2701 aaggcggagc ggggggggct tagcgagctt gacaaggcgg ggttcatcaa gcgtcagctt
     2761 gtcgagaccc gtcagatcac caaacacgtt gctcagatcc ttgacagccg tatgaacacc
     2821 aagtacgacg agaacgacaa gcttatccgt gaagtcaagg ttatcacgct caagtcgaag
     2881 ctcgtctcgg attttcgtaa ggactttcag ttttacaagg ttcgggaaat caacaactac
     2941 caccacgcac acgacgctta ccttaacgca gttgtcggga ccgcgcttat caaaaagtac
     3001 cccaagcttg agtccgaatt cgtttacggg gactacaagg tctacgacgt ccggaagatg
     3061 atcgcgaaat cggaacagga gattgggaag gctacggcta agtacttttt ctactcgaac
     3121 atcatgaact tttttaagac cgagatcacc cttgctaacg gggaaatccg taagcgtccc
     3181 cttatcgaga ccaacggaga gaccggggag atcgtttggg acaagggacg tgacttcgca
     3241 accgttcgta aggttctttc catgcctcag gttaacatcg ttaaaaagac cgaggttcag
     3301 accggggggt tttccaaaga atcgatcctc cccaagagga actccgataa acttatcgcg
     3361 cggaagaagg attgggaccc caagaagtac ggggggttcg acagccccac ggttgcgtac
     3421 tcggttcttg ttgtcgcgaa agttgagaag ggaaagtcca aaaaactcaa gtccgtcaag
     3481 gaactccttg gaatcaccat catggagagg agctccttcg agaagaaccc tatcgatttc
     3541 ctcgaggcga aggggtacaa agaggtcaag aaggatctca tcatcaagct tcccaagtac
     3601 agccttttcg aactcgagaa cgggcggaag cgaatgttgg cgagcgcagg ggagctccag
     3661 aaggggaacg aattggcgtt gccctccaag tacgttaact tcctctacct cgcgagccac
     3721 tacgagaaac ttaaagggag ccccgaggac aacgagcaga agcagctttt cgttgaacag
     3781 cacaaacact acctcgacga aatcatcgaa cagatcagcg agttctcgaa acgggtcatc
     3841 ctcgctgacg caaacctcga caaggtcctt agcgcttaca acaagcacag ggataagccc
     3901 attcgggagc aggctgagaa catcatccac ctcttcacgc ttacgaacct tggggcgcct
     3961 gcggcgttca agtacttcga caccaccatc gaccggaagc gttacacgtc gaccaaggag
     4021 gtcctcgacg caacccttat ccaccagtcc atcaccgggc tttacgagac ccgtatcgac
     4081 cttagccagc ttggggggga tcaccatcac catcaccatt agcggccgca taatgatgtg
     4141 ttatcattga tgcgaggtcg cctatacctc cccgcttcgg cggggttttt tcccgggttt
     4201 acactttatg cttccggctc gtataatgga tggcaggtgc cccaacccta cccacctgct
     4261 agagttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
     4321 agtggcaccg agtcggtgct ttgatccggt ggatgacctt ttgaatgacc tttaatagat
     4381 tatattacta attaattggg gaccctagag gtcccctttt ttattttaaa aattttttca
     4441 caaaacggtt tacaagcata aagcttgctc aatcaatcac cgtcgacctc gtctccccgc
     4501 ttcggcgggg ttttttccta ggttgtgtct caaaatctct gatgttacat tgcacaagat
     4561 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     4621 gttatgagcc atattcagcg tgaaacgagc tgtagccgtc cgcgtctgaa cagcaacatg
     4681 gatgcggatc tgtatggcta taaatgggcg cgtgataacg tgggtcagag cggcgcgacc
     4741 atttatcgtc tgtatggcaa accggatgcg ccggaactgt ttctgaaaca tggcaaaggc
     4801 agcgtggcga acgatgtgac cgatgaaatg gtgcgtctga actggctgac cgaatttatg
     4861 ccgctgccga ccattaaaca ttttattcgc accccggatg atgcgtggct gctgaccacc
     4921 gcgattccgg gcaaaaccgc gtttcaggtg ctggaagaat atccggatag cggcgaaaac
     4981 attgtggatg cgctggccgt gtttctgcgt cgtctgcata gcattccggt gtgcaactgc
     5041 ccgtttaaca gcgatcgtgt gtttcgtctg gcccaggcgc agagccgtat gaacaacggc
     5101 ctggtggatg cgagcgattt tgatgatgaa cgtaacggct ggccggtgga acaggtgtgg
     5161 aaagaaatgc ataaactgct gccgtttagc ccggatagcg tggtgaccca cggcgatttt
     5221 agcctggata acctgatttt cgatgaaggc aaactgattg gctgcattga tgtgggccgt
     5281 gtgggcattg cggatcgtta tcaggatctg gccattctgt ggaactgcct gggcgaattt
     5341 agcccgagcc tgcaaaaacg tctgtttcag aaatatggca ttgataatcc ggatatgaac
     5401 aaactgcaat ttcatctgat gctggatgaa tttttctaat aattaattgg accgcggtcc
     5461 gagagacgca tggcagtatc tcagactgtg tatcggcacg tctcactact taccggcgcg
     5521 gcagcgttac ccgtgtcggc ggctccaacg gctcgccatc gtccagaaaa cacggctcat
     5581 cgggcatcgg caggcgctgc tgcccgcgcc gttcccattc ctccgtttcg gtcaaggctg
     5641 gcaggtctgg ttccatgccc ggaatgccgg gctggctggg cggctcctcg ccggggccgg
     5701 tcggtagttg ctgctcgccc ggatacaggg tcgggatgcg gcgcaggtcg ccatgcccca
     5761 acagcgattc gtcctggtcg tcgtgatcaa ccaccacggc ggcactgaac accgacaggc
     5821 gcaactggtc gcggggctgg ccccacgcca cgcggtcatt gaccacgtag gccgacacgg
     5881 tgccggggcc gttgagcttc acgacggaga tccagcgctc ggccaccaag tccttgactg
     5941 cgtattggac cgtccgcaaa gaacgtccga tgagcttgga aagtgttttc tggctgacca
     6001 ccacggcgtt ctggtggccc atctgcgcca cgaggtgatg cagcagcatt gccgccgtgg
     6061 gtttcctcgc aataagcccg gcccacgcct catgcgcttt gcgttccgtt tgcacccagt
     6121 gaccgggctt gttcttggct tgaatgccga tttctctgga ctgcgtggcc atgcttatct
     6181 ccatgcggta ggggtgccgc acggttgcgg caccatgcgc aatcagctgc aacttttcgg
     6241 cagcgcgaca acaattatgc gttgcgtaaa agtggcagtc aattacagat tttctttaac
     6301 ctacgcaatg agctattgcg gggggtgccg caatgagctg ttgcgtaccc ccctttttta
     6361 agttgttgat ttttaagtct ttcgcatttc gccctatatc tagttctttg gtgcccaaag
     6421 aagggcaccc ctgcggggtt cccccacgcc ttcggcgcgg ctccccctcc ggcaaaaagt
     6481 ggcccctccg gggcttgttg atcgactgcg cggccttcgg ccttgcccaa ggtggcgctg
     6541 cccccttgga acccccgcac tcgccgccgt gaggctcggg gggcaggcgg gcgggcttcg
     6601 cccttcgact gcccccactc gcataggctt gggtcgttcc aggcgcgtca aggccaagcc
     6661 gctgcgcggt cgctgcgcga gccttgaccc gccttccact tggtgtccaa ccggcaagcg
     6721 aagcgcgcag gccgcaggcc ggaggctttt ccccagagaa aattaaaaaa attgatgggg
     6781 caaggccgca ggccgcgcag ttggagccgg tgggtatgtg gtcgaaggct gggtagccgg
     6841 tgggcaatcc ctgtggtcaa gctcgtgggc aggcgcagcc tgtccatcag cttgtccagc
     6901 agggttgtcc acgggccgag cgaagcgagc cagccggtgg ccgctcgcgg ccatcgtcca
     6961 catatccacg ggctggcaag ggagcgcagc gaccgcgcag ggcgaagccc ggagagcaag
     7021 cccgtagggg gggcgcgccc agctgtctag ggcggcggat ttgtcctact caggagagcg
     7081 ttcaccgaca aacaacagat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt
     7141 atttgatgcc tttaattaaa gcggataaca atttcacaca ggaggccgtt tacactttat
     7201 gcttccggct cgtataatgt gtggggaaac cacaacggtt tccctctaga aataattttg
     7261 tttaactata agaaggagat atacatatgt cctctggtta attgaggcct gagtataagg
     7321 tgacttatac ttgtaatcta tctaaacggg gaacctctct agtagacaat cccgtgctaa
     7381 attgatacca gcatcgtctt gatgcccttg gcagcataaa tgcctaacga ctatcccttt
     7441 ggggagtagg gtcaagtgac tcgaaacgat agacaacttg ctttaacaag ttggagatat
     7501 agtctgctct gcatggtgac atgcagctgg atataattcc ggggtaagat taacgacctt
     7561 atctgaacat aatgcta
//
