LOCUS       sequence-260955-        9018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RSF1010-based broad host-range plasmid with RK2 bom site and pUC 
            origin expressing YFP and Sp/Sm resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9018)
  AUTHORS   Bishe B, Taton A, Golden JW
  TITLE     Modification of RSF1010-Based Broad-Host-Range Plasmids for Improved
            Conjugation and Cyanobacterial Bioprospecting.
  JOURNAL   iScience. 2019 Oct 25;20:216-228. doi: 10.1016/j.isci.2019.09.002. 
            Epub 2019 Sep 10.
  PUBMED    31585408
REFERENCE   2  (bases 1 to 9018)
  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: "iScience.";
            date: "2019-10-25"; volume: "20:216-228. doi"; pages: " 
            10.1016/j.isci.2019.09.002. Epub 2019 Sep 10"
COMMENT     Sequence Label: pAM5411
FEATURES             Location/Qualifiers
     source          1..9018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(397..1248)
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(1235..2074)
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             complement(2585..3556)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELSERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     oriT            4795..4882
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      5224..5618
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     terminator      complement(5781..5824)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     oriT            5920..6029
                     /note="incP origin of transfer"
     rep_origin      6206..6794
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6695..6714
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6904..7692)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        join(8128..8133,8134..8149,8150..8156)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             join(8187..8189,8190..8192,8193..8906)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVRFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8493..8512)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8840..8861
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(8922..8946)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      8956..8983
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tctagaacaa tggaagtcct cgtctcgccc tcgaattagc ccgcttaatg agcgggcttt
       61 ttttgaatta attctcgcga gctggcacga caggtttccc gactggaaag cgggcagtga
      121 gcgcaacgca attaatgtaa gttagcgcga attgcaagct ggccgacgcg ctgggctacg
      181 tcttgctggc gttcgggagc agaagagcat acatctggaa gcaaagccag gaaagcggcc
      241 tatggagctg tgcggcagcg ctcagtaggc aatttttcaa aatattgtta agccttttct
      301 gagcatggta tttttcatgg tattaccaat tagcaggaaa ataagccatt gaatataaaa
      361 gataaaaatg tcttgtttac aatagagtgg ggggggtcag cctgccgcct tgggccgggt
      421 gatgtcgtac ttgcccgccg cgaactcggt taccgtccag cccagcgcga ccagctccgg
      481 caacgcctcg cgcacccgct ggcggcgctt gcgcatggtc gaaccactgg cctctgacgg
      541 ccagacatag ccgcacaagg tatctatgga agccttgccg gttttgccgg ggtcgatcca
      601 gccacacagc cgctggtgca gcaggcgggc ggtttcgctg tccagcgccc gcacctcgtc
      661 catgctgatg cgcacatgct ggccgccacc catgacggcc tgcgcgatca aggggttcag
      721 ggccacgtac aggcgcccgt ccgcctcgtc gctggcgtac tccgacagca gccgaaaccc
      781 ctgccgcttg cggccattct gggcgatgat ggataccttc caaaggcgct cgatgcagtc
      841 ctgtatgtgc ttgagcgccc caccactatc gacctctgcc ccgatttcct ttgccagcgc
      901 ccgatagcta cctttgacca ccatggcatc agcggtgacg gcctcccact tgggttccag
      961 gaacagccgg agctgccgtc cgccttcggt cttgggttcc gggccaagca ctaggccatt
     1021 aggcccagcc atggccacca gcccttgcag gatgcgcaga tcatcagcgc ccagcggctc
     1081 cgggccgctg aactcgatcc gcttgccgtc gccgtagtca tacgtcacgt ccagcttgct
     1141 gcgcttgcgc tcgccccgct tgagggcacg gaacaggccg ggggccagac agtgcgccgg
     1201 gtcgtgtcgg acgtggctga ggctgtgctt gttcttaggc ttcaccacgg ggcaccccct
     1261 tgctcttgcg ctgcctctcc agcacggcgg gcttgagcac cccgccgtca tgccgcctga
     1321 accaccgatc agcgaacggt gcgccatagt tggccttgct cacaccgaag cggacgaaga
     1381 accggcgctg gtcgtcgtcc acaccccatt cctcggcctc ggcgctggtc atgctcgaca
     1441 ggtaggactg ccagcggatg ttatcgacca gtaccgagct gccccggctg gcctgctgct
     1501 ggtcgcctgc gcccatcatg gccgcgccct tgctggcatg gtgcaggaac acgatagagc
     1561 acccggtatc ggcggcgatg gcctccatgc gaccgatgac ctgggccatg gggccgctgg
     1621 cgttttcttc ctcgatgtgg aaccggcgca gcgtgtccag caccatcagg cggcggccct
     1681 cggcggcgcg cttgaggccg tcgaaccact ccggggccat gatgttgggc aggctgccga
     1741 tcagcggctg gatcagcagg ccgtcagcca cggcttgccg ttcctcggcg ctgaggtgcg
     1801 ccccaagggc gtgcaggcgg tgatgaatgg cggtgggcgg gtcttcggcg ggcaggtaga
     1861 tcaccgggcc ggtgggcagt tcgcccacct ccagcagatc cggcccgcct gcaatctgtg
     1921 cggccagttg cagggccagc atggatttac cggcaccacc gggcgacacc agcgccccga
     1981 ccgtaccggc caccatgttg ggcaaaacgt agtccagcgg tggcggcgct gctgcgaacg
     2041 cctccagaat attgataggc ttatgggtag ccattgattg cctcctttgc aggcagttgg
     2101 tggttaggcg ctggcggggt cactaccccc gccctgcgcc gctctgagtt cttccaggca
     2161 ctcgcgcagc gcctcgtatt cgtcgtcggt cagccagaac ttgcgctgac gcatcccttt
     2221 ggccttcatg cgctcggcat atcgcgcttg gcgtacagcg tcagggctgg ccagcaggtc
     2281 gccggtctgc ttgtcctttt ggtctttcat atcagtcacc gagaaacttg ccggggccga
     2341 aaggcttgtc ttcgcggaac aaggacaagg tgcagccgtc aaggttaagg ctggccatat
     2401 cagcgactga aaagcggcca gcctcggcct tgtttgacgt ataaccaaag ccaccgggca
     2461 accaatagcc cttgtcactt ttgatcaggt agaccgaccc tgaagcgctt ttttcgtatt
     2521 ccataaaacc cccttctgtg cgtgagtact catagtataa caggcgtgag taccaacgca
     2581 agcactacat gctgaaatct ggcccgcccc tgtccatgcc tcgctggcgg ggtgccggtg
     2641 cccgtgccag ctcggcccgc gcaagctgga cgctgggcag acccatgacc ttgctgacgg
     2701 tgcgctcgat gtaatccgct tcgtggccgg gcttgcgctc tgccagcgct gggctggcct
     2761 cggccatggc cttgccgatt tcctcggcac tgcggccccg gctggccagc ttctgcgcgg
     2821 cgataaagtc gcacttgctg aggtcatcac cgaagcgctt gaccagcccg gccatctcgc
     2881 tgcggtactc gtccagcgcc gtgcgccggt ggcggctaag ctgccgctcg gacagttcga
     2941 ggctggccag cctgcgggcc ttctcctgct gccgctgggc ctgctcgatc tgctggccag
     3001 cctgctgcac cagcgccggg ccagcggtgg cggtcttgcc cttggattca cgcagcagca
     3061 cccacggctg ataaccggcg cgggtggtgt gcttgtcctt gcggttggtg aagcccgcca
     3121 agcggccata gtggcggctg tcggcgctgg ccgggtcggc gtcgtactcg ctggccagcg
     3181 tccgggcaat ctgcccccga agttcaccgc ctgcggcgtc ggccaccttg acccatgcct
     3241 gatagttctt cgggctggtt tccactacta gggcaggctc ccggccctcg gctttcatgt
     3301 catccaggtc aaactcgctg aggtcgtcca ccagcaccag accatgccgc tcctgctcgg
     3361 cgggcctgat atacacgtca ttgccctggg cattcatccg cttgagccat ggcgtgttct
     3421 ggagcacttc ggcggctgac cattcccggt tcatcatctg gccggtggtg gcgtccctga
     3481 cgccgatatc gaagcgctca cagcccatgg ccttgagctg tcggcctatg gcctgcaaag
     3541 tcctgtcgtt cttcatcggg ccaccaagcg cagccagatc gagccgtcct cggttgtcag
     3601 tggcgtcagg tcgagcaaga gcaacgatgc gatcagcagc accaccgtag gcatcatgga
     3661 agccagcatc acggttagcc atagcttcca gtgccacccc cgcgacgcgc tccgggcgct
     3721 ctgcgcggcg ctgctcacct cggcggctac ctcccgcaac tctttggcca gctccaccca
     3781 tgccgcccct gtctggcgct gggctttcag ccactccgcc gcctgcgcct cgctggcctg
     3841 ctgggtctgg ctcatgacct gccgggcttc gtcggccagt gtcgccatgc tctgggccag
     3901 cggttcgatc tgctccgcta actcgttgat gcctctggat ttcttcactc tgtcgattgc
     3961 gttcatggtc tattgcctcc cggtattcct gtaagtcgat gatctgggcg ttggcggtgt
     4021 cgatgttcag ggccacgtct gcccggtcgg tgcggatgcc ccggccttcc atctccacca
     4081 cgttcggccc caggtgaaca ccgggcaggc gctcgatgcc ctgcgcctca agtgttctgt
     4141 ggtcaatgcg ggcgtcgtgg cctgcccgct ctaatgcccg gttggcatgg tcggcccatg
     4201 cctcgcgggt ctgctcaagc catgccttgg gcttgagcgc ttcggtcttc tgtgccccgc
     4261 ccttctccgg ggtcttgccg ttgtaccgct tgaaccactg agcggcgggc cgctcgatgc
     4321 cgtcattgat ccgctcggag atcatcaggt ggcagtgcgg gttctcgccg ccaccggcat
     4381 ggatggccag cgtatacggc aggcgctcgg caccggtcag gtgctgggcg aactcggacg
     4441 ccagcgcctt ctgctggtcg agggtcagct cgaccggcag ggcaaattcg acctccttga
     4501 acagccgccc attggcgcgt tcatacaggt cggcagcatc ccagtagtcg gcgggccgct
     4561 cgacgaactc cggcatgtgc ccggattcgg cgtgcaagac ttcatccatg tcgcgggcat
     4621 acttgccttc gcgctggatg aagtcggcct tggccctggc cgattggccg cccgacctgc
     4681 tgccggtttt cgccgtaagg tgataaatcg ccatgctgcc tcgctgttgc ttttgctttt
     4741 cggctccatg caatggccct cggagagcgc accgcccgaa gggtggccgt taggccagtt
     4801 tctcgaagag aaaccggtaa gtgcgccctc ccctacaaag tagggtcggg attgccgccg
     4861 ctgtgcctcc atgatagcct acgagacagc acattaacaa tggggtgtca agatggttaa
     4921 ggggagcaac aaggcggcgg atcggctggc caagctcgaa gaacaacgag cgcgaatcaa
     4981 tgccgaaatt cagcgggtgc gggcaaggga acagcagcaa gagcgcaaga acgaaacaag
     5041 gcgcaaggtg ctggtggggg ccatgatttt ggccaaggtg aacagcagcg agtggccgga
     5101 ggatcggctc atggcggcaa tggatgcgta ccttgaacgc gaccacgacc gcgccttgtt
     5161 cggtctgccg ccacgccaga aggatgagcc gggctgaatg atcgaccgag acaggccctg
     5221 cggggctgca cacgcgcccc cacccttcgg gtagggggaa aggccgctaa agcggctaaa
     5281 agcgctccag cgtatttctg cggggtttgg tgtggggttt agcgggcttt gcccgccttt
     5341 ccccctgccg cgcagcggtg gggcggtgtg tagcctagcg cagcgaatag accagctatc
     5401 cggcctctgg ccgggcatat tgggcaaggg cagcagcgcc ccacaagggc gctgataacc
     5461 gcgcctagtg gattattctt agataatcat ggatggattt ttccaacacc ccgccagccc
     5521 ccgcccctgc tgggtttgca ggtttggggg cgtgacagtt attgcagggg ttcgtgacag
     5581 ttattgcagg ggggcgtgac agttattgca ggggttcgtg acagttagta cgggagtgac
     5641 gggcactggc tggcaatgtc tagcaacggc aggcatttcg gctgagggta aaagaacttt
     5701 ccgctaagcg atagactgta tgtaaacaca gtattgcaag gacgcggaac atgcctcatg
     5761 tggcggccct cgcgagaatt aattcagata aaaaaaatcc ttagctttcg ctaaggatga
     5821 tttctagcga tgaccctgct gattggttcg ctgaccgtag actggttggc ttggtttcat
     5881 cagccatccg cttgccctca tctgttacgc cggcggtagc cggccagcct cgcagagcag
     5941 gattcccgtt gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa cacccgctcg
     6001 cgggtgggcc tacttcacct atcctgcccg gctgacgccg ttggatacac caaggaaagt
     6061 ctacacgaac cctttggcaa aatcctgtat atcgtgcgag gatctaggtg aagatccttt
     6121 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6181 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6241 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6301 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6361 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6421 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6481 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6541 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6601 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6661 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6721 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6781 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc tttcatcgcg
     6841 ccgcgcgcgc ggatgctagc ggcgcgccaa aaaaaagccc gctcattagg cgggctggcc
     6901 ggccttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg
     6961 atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag
     7021 tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt
     7081 cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt
     7141 tcgctcatcg ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc
     7201 ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa
     7261 ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt cgatcgtggc
     7321 tggctcgaag atacctgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg
     7381 cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc
     7441 gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc
     7501 tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg
     7561 tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt acggccagca acgtcggttc
     7621 gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt cggcgatcac
     7681 cgcttccctc atgatgttta actttgtttt agggcgactg ccctgctgcg taacatcgtt
     7741 gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct tggatgcccg
     7801 aggcatagac tgtaccccaa aaaaacagtc ataacaagcc atgaaaaccg ccactgcgcc
     7861 gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca tacgctactt
     7921 gcattacagc ttacgaaccg aacaggctta tgtccactgg gttcgtgcct tcatccgttt
     7981 ccacggtgtg cgtcacccgg caaccttggg cagcagcgaa gtcgaggcat ttctgtcctg
     8041 gctggcgaac gagcgcaagg tttcggtctc cacgcattcg aaaccggtat cccgccgccg
     8101 ccgccggata ccggtttcga attgagattg acaattaatc atcggctcgt ataatggtac
     8161 cctatttaaa taaaggaggt cttaagatgg tgagcaaggg cgaggagctg ttcaccgggg
     8221 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     8281 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     8341 gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc ctgcagtgct
     8401 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     8461 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     8521 aggtgaggtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     8581 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     8641 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     8701 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     8761 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc
     8821 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     8881 tcggcatgga cgagctgtac aagtaaaggg tgggcgcgcc gacccagctt tcttgtacaa
     8941 actcggccct gcaggagaag gccatcctga cggatggcct ttttgaagct ttctagaatc
     9001 gggggccccc ggggggat
//
