LOCUS       sequence-126476-        8958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RSF1010-based vector with extensive polylinker, and resistance to 
            Strep and Spec. Shuttle vector suitable for transfer to, and 
            replication in, E. coli and Anabaena (Nostoc) sp. strain PCC 7120..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8958)
  AUTHORS   Wolk CP, Fan Q, Zhou R, Huang G, Lechno-Yossef S, Kuritz T, Wojciuch
            E
  TITLE     Paired cloning vectors for complementation of mutations in the 
            cyanobacterium Anabaena sp. strain PCC 7120.
  JOURNAL   Arch Microbiol. 2007 Dec;188(6):551-63. Epub 2007 Jul 17.
  PUBMED    17639350
REFERENCE   2  (bases 1 to 8958)
  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: "Arch 
            Microbiol."; date: "2007-12"; volume: "188(6)"; pages: "551-63. Epub
            2007 Jul 17"
COMMENT     Sequence Label: pRL1383a
FEATURES             Location/Qualifiers
     source          1..8958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      155..549
                     /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)"
     oriT            891..978
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             2217..3188
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDAPTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGHDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     CDS             3699..4538
                     /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             4525..5376
                     /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
                     TAECHVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRKRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     misc_feature    5758..6157
                     /label=cos
                     /note="lambda cos site; allows packaging into phage lambda 
                     particles"
     protein_bind    complement(6168..6201)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     6217..6236
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6217..6235
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6398..6415)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6590..6609)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             7447..8238
                     /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"
     terminator      8903..8946
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 tcctggccgc cacatgaggc atgttccgcg tccttgcaat actgtgttta catacagtct
       61 atcgcttagc ggaaagttct tttaccctca gccgaaatgc ctgccgttgc tagacattgc
      121 cagccagtgc ccgtcactcc cgtactaact gtcacgaacc cctgcaataa ctgtcacgcc
      181 cccctgcaat aactgtcacg aacccctgca ataactgtca cgcccccaaa cctgcaaacc
      241 cagcaggggc gggggctggc ggggtgttgg aaaaatccat ccatgattat ctaagaataa
      301 tccactaggc gcggttatca gcgcccttgt ggggcgctgc tgcccttgcc caatatgccc
      361 ggccagaggc cggatagctg gtctattcgc tgcgctaggc tacacaccgc cccaccgctg
      421 cgcggcaggg ggaaaggcgg gcaaagcccg ctaaacccca caccaaaccc cgcagaaata
      481 cgctggagcg cttttagccg ctttagcggc ctttccccct acccgaaggg tgggggcgcg
      541 tgtgcagccc cgcagggcct gtctcggtcg atcattcagc ccggctcatc cttctggcgt
      601 ggcggcagac cgaacaaggc gcggtcgtgg tcgcgttcaa ggtacgcatc cattgccgcc
      661 atgagccgat cctccggcca ctcgctgctg ttcaccttgg ccaaaatcat ggcccccacc
      721 agcaccttgc gccttgtttc gttcttgcgc tcttgctgct gttcccttgc ccgctcccgc
      781 tgaatttcgg cattgattcg cgctcgttgt tcttcgagct tggccagccg atccgccgcc
      841 ttgttgctcc ccttaaccat cttgacaccc cattgttaat gtgctgtctc gtaggctatc
      901 atggaggcac agcggcggca atcccgaccc tactttgtag gggagggcgc acttaccggt
      961 ttctcttcga gaaactggcc taacggccac ccttcgggcg gtgcgctctc cgagggccat
     1021 tgcatggagc cgaaaagcaa aagcaacagc gaggcagcat ggcgatttat caccttacgg
     1081 cgaaaaccgg cagcaggtcg ggcggccaat cggccagggc caaggccgac tacatccagc
     1141 gcgaaggcaa gtatgcccgc gacatggatg aagtcttgca cgccgaatcc gggcacatgc
     1201 cggagttcgt cgagcggccc gccgactact gggatgctgc cgacctgtat gaacgcgcca
     1261 atgggcggct gttcaaggag gtcgaatttg ccctgccggt cgagctgacc ctcgaccagc
     1321 agaaggcgct ggcgtccgag ttcgcccagc acctgaccgg tgccgagcgc ctgccgtata
     1381 cgctggccat ccatgccggt ggcggcgaga acccgcactg ccacctgatg atctccgagc
     1441 ggatcaatga cggcatcgag cggcccgccg ctcagtggtt caagcggtac aacggcaaga
     1501 ccccggagaa gggcggggca cagaagaccg aagcgctcaa gcccaaggca tggcttgagc
     1561 agacccgcga ggcatgggcc gaccatgcca accgggcatt agagcgggct ggccacgacg
     1621 cccgcattga ccacagaaca cttgaggcgc agggcatcga gcgcctgccc ggtgttcacc
     1681 tggggccgaa cgtggtggag atggaaggcc ggggcatccg caccgaccgg gcagacgtgg
     1741 ccctgaacat cgacaccgcc aacgcccaga tcatcgactt acaggaatac cgggaggcaa
     1801 tagaccatga acgcaatcga cagagtgaag aaatccagag gcatcaacga gttagcggag
     1861 cagatcgaac cgctggccca gagcatggcg acactggccg acgaagcccg gcaggtcatg
     1921 agccagacca agcaggccag cgaggcgcag gcggcggagt ggctgaaagc ccagcgccag
     1981 acaggggcgg catgggtgga gctggccaaa gagttgcggg aggtagccgc cgaggtgagc
     2041 agcgccgcgc agagcgcccg gagcgcgtcg cgggggtggc actggaagct atggctaacc
     2101 gtgatgctgg cttccatgat gcctacggtg gtgctgctga tcgcatcgtt gctcttgctc
     2161 gacctgacgc cactgacaac cgaggacggc tcgatctggc tgcgcttggt ggcccgatga
     2221 agaacgacag gactttgcag gccataggcc gacagctcaa ggccatgggc tgtgagcgct
     2281 tcgatatcgg cgtcagggac gcacccaccg gccagatgat gaaccgggaa tggtcagccg
     2341 ccgaagtgct ccagaacacg ccatggctca agcggatgaa tgcccagggc aatgacgtgt
     2401 atatcaggcc cgccgagcag gagcggcatg gtctggtgct ggtggacgac ctcagcgagt
     2461 ttgacctgga tgacatgaaa gccgagggcc gggagcctgc cctggtagtg gaaaccagcc
     2521 cgaagaacta tcaggcatgg gtcaaggtgg ccgacgccgc aggcggtgaa cttcgggggc
     2581 agattgcccg gacgctggcc agcgagtacg acgccgaccc ggccagcgcc gacagccgcc
     2641 actatggccg cttggcgggc ttcaccaacc gcaaggacaa gcacaccacc cgcgccggtt
     2701 atcagccgtg ggtgctgctg cgtgaatcca agggcaagac cgccaccgct ggcccggcgc
     2761 tggtgcagca ggctggccag cagatcgagc aggcccagcg gcagcaggag aaggcccgca
     2821 ggctggccag cctcgaactg cccgagcggc agcttagccg ccaccggcgc acggcgctgg
     2881 acgagtaccg cagcgagatg gccgggctgg tcaagcgctt cggtcatgac ctcagcaagt
     2941 gcgactttat cgccgcgcag aagctggcca gccggggccg cagtgccgag gaaatcggca
     3001 aggccatggc cgaggccagc ccagcgctgg cagagcgcaa gcccggccac gaagcggatt
     3061 acatcgagcg caccgtcagc aaggtcatgg gtctgcccag cgtccagctt gcgcgggccg
     3121 agctggcacg ggcaccggca ccccgccagc gaggcatgga caggggcggg ccagatttca
     3181 gcatgtagtg cttgcgttgg tactcacgcc tgttatacta tgagtactca cgcacagaag
     3241 ggggttttat ggaatacgaa aaaagcgctt cagggtcggt ctacctgatc aaaagtgaca
     3301 agggctattg gttgcccggt ggctttggtt atacgtcaaa caaggccgag gctggccgct
     3361 tttcagtcgc tgatatggcc agccttaacc ttgacggctg caccttgtcc ttgttccgcg
     3421 aagacaagcc tttcggcccc ggcaagtttc tcggtgactg atatgaaaga ccaaaaggac
     3481 aagcagaccg gcgacctgct ggccagccct gacgctgtac gccaagcgcg atatgccgag
     3541 cgcatgaagg ccaaagggat gcgtcagcgc aagttctggc tgaccgacga cgaatacgag
     3601 gcgctgcgcg agtgcctgga agaactcaga gcggcgcagg gcgggggtag tgaccccgcc
     3661 agcgcctaac caccaactgc ctgcaaagga ggcaatcaat ggctacccat aagcctatca
     3721 atattctgga ggcgttcgca gcagcgccgc caccgctgga ctacgttttg cccaacatgg
     3781 tggccggtac ggtcggggcg ctggtgtcgc ccggtggtgc cggtaaatcc atgctggccc
     3841 tgcaactggc cgcacagatt gcaggcgggc cggatctgct ggaggtgggc gaactgccca
     3901 ccggcccggt gatctacctg cccgccgaag acccgcccac cgccattcat caccgcctgc
     3961 acgcccttgg ggcgcacctc agcgccgagg aacggcaagc cgtggctgac ggcctgctga
     4021 tccagccgct gatcggcagc ctgcccaaca tcatggcccc ggagtggttc gacggcctca
     4081 agcgcgccgc cgagggccgc cgcctgatgg tgctggacac gctgcgccgg ttccacatcg
     4141 aggaagaaaa cgccagcggc cccatggccc aggtcatcgg tcgcatggag gccatcgccg
     4201 ccgataccgg gtgctctatc gtgttcctgc accatgccag caagggcgcg gccatgatgg
     4261 gcgcaggcga ccagcagcag gccagccggg gcagctcggt actggtcgat aacatccgct
     4321 ggcagtccta cctgtcgagc atgaccagcg ccgaggccga ggaatggggt gtggacgacg
     4381 accagcgccg gttcttcgtc cgcttcggtg tgagcaaggc caactatggc gcaccgttcg
     4441 ctgatcggtg gttcaggcgg catgacggcg gggtgctcaa gcccgccgtg ctggagaggc
     4501 agcgcaagag caagggggtg ccccgtggtg aagcctaaga acaagcacag cctcagccac
     4561 gtccggcacg acccggcgca ctgtctggcc cccggcctgt tccgtgccct caagcggggc
     4621 gagcgcaagc gcagcaagct ggacgtgacg tatgactacg gcgacggcaa gcggatcgag
     4681 ttcagcggcc cggagccgct gggcgctgat gatctgcgca tcctgcaagg gctggtggcc
     4741 atggctgggc ctaatggcct agtgcttggc ccggaaccca agaccgaagg cggacggcag
     4801 ctccggctgt tcctggaacc caagtgggag gccgtcaccg ctgaatgcca tgtggtcaaa
     4861 ggtagctatc gggcgctggc aaaggaaatc ggggcagagg tcgatagtgg tggggcgctc
     4921 aagcacatac aggactgcat cgagcgcctt tggaaggtat ccatcatcgc ccagaatggc
     4981 cgcaagcggc aggggtttcg gctgctgtcg gagtacgcca gcgacgaggc ggacgggcgc
     5041 ctgtacgtgg ccctgaaccc cttgatcgcg caggccgtca tgggtggcgg ccagcatgtg
     5101 cgcatcagca tggacgaggt gcgggcgctg gacagcgaaa ccgcccgcct gctgcaccag
     5161 cggctgtgtg gctggatcga ccccggcaaa accggcaagg cttccataga taccttgtgc
     5221 ggctatgtct ggccgtcaga ggccagtggt tcgaccatgc gcaagcgccg caagcgggtg
     5281 cgcgaggcgt tgccggagct ggtcgcgctg ggctggacgg taaccgagtt cgcggcgggc
     5341 aagtacgaca tcacccggcc caaggcggca ggctgacccc ccccactcta ttgtaaacaa
     5401 gacattttta tcttttatat tcaatggctt attttcctgc taattggtaa taccatgaaa
     5461 aataccatgc tcagaaaagg cttaacaata ttttgaaaaa ttgcctactg agcgctgccg
     5521 cacagctcca taggccgctt tcctggcttt gcttccagat gtatgctctt ctgctcccga
     5581 acgccagcaa gacgtagccc agcgcgtcgg ccagcttgca attcgcgcta acttacatta
     5641 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctcgcgaga
     5701 attaattcaa aaaaagcccg ctcattaggc gggctaattc gagggcgaga cgaggacttc
     5761 cattgttcat tccacggaca aaaacagaga aaggaaacga cagaggccaa aaagcctcgc
     5821 tttcagcacc tgtcgtttcc tttcttttca gagggtattt taaataaaaa cattaagtta
     5881 tgacgaagaa gaacggaaac gccttaaacc ggaaaatttt cataaatagc gaaaacccgc
     5941 gaggtcgccg ccccgtaacc tgtcggatca ccggaaagga cccgtaaagt gataatgatt
     6001 atcatctaca tatcacaacg tgcgtggagg ccatcaaacc acgtcaaata atcaattatg
     6061 acgcaggtat cgtattaatt gatctgcatc aacttaacgt aaaaacaact tcagacaata
     6121 caaatcagcg acactgaata cggggcaacc tcatgtcctc atcgctaata acttcgtata
     6181 gcatacatta tacgaagtta tattcgatgc ggccgctaat acgactcact atagggagaa
     6241 gcttgcatgc ctgcaggtcg actctagggg taccgagctc gaattcatcg atctcgtaac
     6301 ccgggcccgc ggatgcatat gatcagatct taaggcctag gtctagacgt cgtacgtgca
     6361 ctcgagcccg ggcgcgccgg accggccggc agggatccta tagtgtcacc taaatcgtat
     6421 gcggccgctc gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt
     6481 attttttgag ttatcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac
     6541 tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca
     6601 gtcagttgct caatgtacct ataaccagac cgttcagaaa ataaaaaagg ggacctctag
     6661 ggtccccaat taattagtaa tataatctat taaaggtcat tcaaaaggtc atccaccgga
     6721 tcaattcccc tgctcgcgca ggctgggtgc caagctctcg ggtaacatca aggcccgatc
     6781 cttggagccc ttgccctccc gcacgatgat cgtgccgtga tcgaaatcca gatccttgac
     6841 ccgcagttgc aaaccctcac tgatccgccc gttccataca gaagctgggc gaacaaacga
     6901 tgctcgcctt ccagaaaacc gaggatgcga accacttcat ccggggtcag caccaccggc
     6961 aagcgccgcg acggccgagg tcttccgatc tcctgaagcc agggcagatc cgtgcacagc
     7021 accttgccgt agaagaacag caaggccgcc aatgcctgac gatgcgtgga gaccgaaacc
     7081 ttgcgctcgt tcgccagcca ggacagaaat gcctcgactt cgctgctgcc caaggttgcc
     7141 gggtgacgca caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg
     7201 gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc
     7261 gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt
     7321 ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt
     7381 ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta
     7441 aacatcatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc
     7501 gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg
     7561 gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt
     7621 gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga
     7681 gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg
     7741 tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt
     7801 gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca
     7861 agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct
     7921 gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac
     7981 tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta
     8041 accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc
     8101 cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc
     8161 ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc
     8221 aaggtagtcg gcaaataatg tctaacaatt cgttcaagcc gacgccgctt cgcggcgcgg
     8281 cttaactcaa gcgttagatg cactaagcac ataattgctc acagccaaac tatcaggtca
     8341 agtctgcttt tattattttt aagcgtgcat aataagccct acacaaattg ggagatatat
     8401 catgaaaggc tggctttttc ttgttatcgc aatagttggc gaagtaatcg caacatccgc
     8461 attaaaatct agcgagggct ttactaagct gatccggtgg atgacctttt gaatgacctt
     8521 taatagatta tattactaat taattgggga ccctagaggt cccctttttt attttaacaa
     8581 agaatacaag aaaatattta caaaaaatca atttaacaat tccttaaaac atgcaggaat
     8641 tgacgattta aacaatatta gctttgaaca attcttatct cttttcaata gctataaatt
     8701 atttaataag taagttaagg gatgcaggtc gagtacccgt acaagcctaa ttgtgtagca
     8761 tctggcttac tgaagcagac cctatcatct ctctcgtaaa ctgccgtcag agtcggtttg
     8821 gttggacgaa ccttctgagt ttctggtaac gccgtcccgc acccggaaat ggtcagcgaa
     8881 ccaatcagca gggtcatcgc tagaaatcat ccttagcgaa agctaaggat tttttttatc
     8941 tgaattaatt ctcgcgag
//
