LOCUS       sequence-126468-        8416 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RSF1010-based vector with extensive polylinker, and resistance to Cm
            and Em. 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 8416)
  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 8416)
  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: pRL1342
FEATURES             Location/Qualifiers
     source          1..8416
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     8..27
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8..26
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(189..206)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             315..974
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(381..400)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2152..2195
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     rep_origin      2362..2756
                     /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            3098..3185
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             4424..5395
                     /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
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     CDS             5906..6745
                     /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             6732..7583
                     /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
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     misc_feature    7965..8364
                     /label=cos
                     /note="lambda cos site; allows packaging into phage lambda 
                     particles"
     protein_bind    complement(8375..8408)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 cggccgctaa tacgactcac tatagggaga agcttgcatg cctgcaggtc gactctaggg
       61 gtaccgagct cgaattcatc gatctcgtaa cccgggcccg cggatgcata tgatcagatc
      121 ttaaggccta ggtctagacg tcgtacgtgc actcgagccc gggcgcgccg gaccggccgg
      181 cagggatcct atagtgtcac ctaaatcgta tgcggccgct cgtaagaggt tccaactttc
      241 accataatga aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag
      301 ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat
      361 ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga
      421 ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt
      481 atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg
      541 caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc
      601 atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt
      661 ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta
      721 aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt
      781 ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat
      841 attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt
      901 gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc
      961 agggcggggc gtaatttttt taaggcagtt attggtgccc ttaaacgcct ggtgctacgc
     1021 ctgaataagt gataataagc ggatgaatgg cagaaattcg atgaattcgc gtgctataat
     1081 tatactaatt ttataaggag gaaaaaatat gggcattttt agtatttttg taatcagcac
     1141 agttcattat caaccaaaca aaaaataagt ggttataatg aatcgttaat aagcaaaatt
     1201 catataacca aattaaagag ggttataatg aacgagaaaa atataaaaca cagtcaaaac
     1261 tttattactt caaaacataa tatagataaa ataatgacaa atataagatt aaatgaacat
     1321 gataatatct ttgaaatcgg ctcaggaaaa ggccatttta cccttgaatt agtaaagagg
     1381 tgtaatttcg taactgccat tgaaatagac cataaattat gcaaaactac agaaaataaa
     1441 cttgttgatc acgataattt ccaagtttta aacaaggata tattgcagtt taaatttcct
     1501 aaaaaccaat cctataaaat atatggtaat ataccttata acataagtac ggatataata
     1561 cgcaaaattg tttttgatag tatagctaat gagatttatt taatcgtgga atacgggttt
     1621 gctaaaagat tattaaatac aaaacgctca ttggcattac ttttaatggc agaagttgat
     1681 atttctatat taagtatggt tccaagagaa tattttcatc ctaaacctaa agtgaatagc
     1741 tcacttatca gattaagtag aaaaaaatca agaatatcac acaaagataa acaaaagtat
     1801 aattatttcg ttatgaaatg ggttaacaaa gaatacaaga aaatatttac aaaaaatcaa
     1861 tttaacaatt ccttaaaaca tgcaggaatt gacgatttaa acaatattag ctttgaacaa
     1921 ttcttatctc ttttcaatag ctataaatta tttaataagt aagttaaggg atgcaggtcg
     1981 agtacccgta caagcctaat tgtgtagcat ctggcttact gaagcagacc ctatcatctc
     2041 tctcgtaaac tgccgtcaga gtcggtttgg ttggacgaac cttctgagtt tctggtaacg
     2101 ccgtcccgca cccggaaatg gtcagcgaac caatcagcag ggtcatcgct agaaatcatc
     2161 cttagcgaaa gctaaggatt ttttttatct gaattaattc tcgcgagtcc tggccgccac
     2221 atgaggcatg ttccgcgtcc ttgcaatact gtgtttacat acagtctatc gcttagcgga
     2281 aagttctttt accctcagcc gaaatgcctg ccgttgctag acattgccag ccagtgcccg
     2341 tcactcccgt actaactgtc acgaacccct gcaataactg tcacgccccc ctgcaataac
     2401 tgtcacgaac ccctgcaata actgtcacgc ccccaaacct gcaaacccag caggggcggg
     2461 ggctggcggg gtgttggaaa aatccatcca tgattatcta agaataatcc actaggcgcg
     2521 gttatcagcg cccttgtggg gcgctgctgc ccttgcccaa tatgcccggc cagaggccgg
     2581 atagctggtc tattcgctgc gctaggctac acaccgcccc accgctgcgc ggcaggggga
     2641 aaggcgggca aagcccgcta aaccccacac caaaccccgc agaaatacgc tggagcgctt
     2701 ttagccgctt tagcggcctt tccccctacc cgaagggtgg gggcgcgtgt gcagccccgc
     2761 agggcctgtc tcggtcgatc attcagcccg gctcatcctt ctggcgtggc ggcagaccga
     2821 acaaggcgcg gtcgtggtcg cgttcaaggt acgcatccat tgccgccatg agccgatcct
     2881 ccggccactc gctgctgttc accttggcca aaatcatggc ccccaccagc accttgcgcc
     2941 ttgtttcgtt cttgcgctct tgctgctgtt cccttgcccg cacccgctga atttcggcat
     3001 tgattcgcgc tcgttgttct tcgagcttgg ccagccgatc cgccgccttg ttgctcccct
     3061 taaccatctt gacaccccat tgttaatgtg ctgtctcgta ggctatcatg gaggcacagc
     3121 ggcggcaatc ccgaccctac tttgtagggg agggcgcact taccggtttc tcttcgagaa
     3181 actggcctaa cggccaccct tcgggcggtg cgctctccga gggccattgc atggagccga
     3241 aaagcaaaag caacagcgag gcagcatggc gatttatcac cttacggcga aaaccggcag
     3301 caggtcgggc ggccaatcgg ccagggccaa ggccgactac atccagcgcg aaggcaagta
     3361 tgcccgcgac atggatgaag tcttgcacgc cgaatccggg cacatgccgg agttcgtcga
     3421 gcggcccgcc gactactggg atgctgccga cctgtatgaa cgcgccaatg ggcggctgtt
     3481 caaggaggtc gaatttgccc tgccggtcga gctgaccctc gaccagcaga aggcgctggc
     3541 gtccgagttc gcccagcacc tgaccggtgc cgagcgcctg ccgtatacgc tggccatcca
     3601 tgccggtggc ggcgagaacc cgcactgcca cctgatgatc tccgagcgga tcaatgacgg
     3661 catcgagcgg cccgccgctc agtggttcaa gcggtacaac ggcaagaccc cggagaaggg
     3721 cggggcacag aagaccgaag cgctcaagcc caaggcatgg cttgagcaga cccgcgaggc
     3781 atgggccgac catgccaacc gggcattaga gcgggctggc cacgacgccc gcattgacca
     3841 cagaacactt gaggcgcagg gcatcgagcg cctgcccggt gttcacctgg ggccgaacgt
     3901 ggtggagatg gaaggccggg gcatccgcac cgaccgggca gacgtggccc tgaacatcga
     3961 caccgccaac gcccagatca tcgacttaca ggaataccgg gaggcaatag accatgaacg
     4021 caatcgacag agtgaagaaa tccagaggca tcaacgagtt agcggagcag atcgaaccgc
     4081 tggcccagag catggcgaca ctggccgacg aagcccggca ggtcatgagc cagacccagc
     4141 aggccagcga ggcgcaggcg gcggagtggc tgaaagccca gcgccagaca ggggcggcat
     4201 gggtggagct ggccaaagag ttgcgggagg tagccgccga ggtgagcagc gccgcgcaga
     4261 gcgcccggag cgcgtcgcgg gggtggcact ggaagctatg gctaaccgtg atgctggctt
     4321 ccatgatgcc tacggtggtg ctgctgatcg catcgttgct cttgctcgac ctgacgccac
     4381 tgacaaccga ggacggctcg atctggctgc gcttggtggc ccgatgaaga acgacaggac
     4441 tttgcaggcc ataggccgac agctcaaggc catgggctgt gagcgcttcg atatcggcgt
     4501 cagggacgcc accaccggcc agatgatgaa ccgggaatgg tcagccgccg aagtgctcca
     4561 gaacacgcca tggctcaagc ggatgaatgc ccagggcaat gacgtgtata tcaggcccgc
     4621 cgagcaggag cggcatggtc tggtgctggt ggacgacctc agcgagtttg acctggatga
     4681 catgaaagcc gagggccggg agcctgccct ggtagtggaa accagcccga agaactatca
     4741 ggcatgggtc aaggtggccg acgccgcagg cggtgaactt cgggggcaga ttgcccggac
     4801 gctggccagc gagtacgacg ccgacccggc cagcgccgac agccgccact atggccgctt
     4861 ggcgggcttc accaaccgca aggacaagca caccacccgc gccggttatc agccgtgggt
     4921 gctgctgcgt gaatccaagg gcaagaccgc caccgctggc ccggcgctgg tgcagcaggc
     4981 tggccagcag atcgagcagg cccagcggca gcaggagaag gcccgcaggc tggccagcct
     5041 cgaactgccc gagcggcagc ttagccgcca ccggcgcacg gcgctggacg agtaccgcag
     5101 cgagatggcc gggctggtca agcgcttcgg tgatgacctc agcaagtgcg actttatcgc
     5161 cgcgcagaag ctggccagcc ggggccgcag tgccgaggaa atcggcaagg ccatggccga
     5221 ggccagccca gcgctggcag agcgcaagcc cggccacgaa gcggattaca tcgagcgcac
     5281 cgtcagcaag gtcatgggtc tgcccagcgt ccagcttgcg cgggccgagc tggcacgggc
     5341 accggcaccc cgccagcgag gcatggacag gggcgggcca gatttcagca tgtagtgctt
     5401 gcgttggtac tcacgcctgt tatactatga gtactcacgc acagaagggg gttttatgga
     5461 atacgaaaaa agcgcttcag ggtcggtcta cctgatcaaa agtgacaagg gctattggtt
     5521 gcccggtggc tttggttata cgtcaaacaa ggccgaggct ggccgctttt cagtcgctga
     5581 tatggccagc cttaaccttg acggctgcac cttgtccttg ttccgcgaag acaagccttt
     5641 cggccccggc aagtttctcg gtgactgata tgaaagacca aaaggacaag cagaccggcg
     5701 acctgctggc cagccctgac gctgtacgcc aagcgcgata tgccgagcgc atgaaggcca
     5761 aagggatgcg tcagcgcaag ttctggctga ccgacgacga atacgaggcg ctgcgcgagt
     5821 gcctggaaga actcagagcg gcgcagggcg ggggtagtga ccccgccagc gcctaaccac
     5881 caactgcctg caaaggaggc aatcaatggc tacccataag cctatcaata ttctggaggc
     5941 gttcgcagca gcgccgccac cgctggacta cgttttgccc aacatggtgg ccggtacggt
     6001 cggggcgctg gtgtcgcccg gtggtgccgg taaatccatg ctggccctgc aactggccgc
     6061 acagattgca ggcgggccgg atctgctgga ggtgggcgaa ctgcccaccg gcccggtgat
     6121 ctacctgccc gccgaagacc cgcccaccgc cattcatcac cgcctgcacg cccttggggc
     6181 gcacctcagc gccgaggaac ggcaagccgt ggctgacggc ctgctgatcc agccgctgat
     6241 cggcagcctg cccaacatca tggccccgga gtggttcgac ggcctcaagc gcgccgccga
     6301 gggccgccgc ctgatggtgc tggacacgct gcgccggttc cacatcgagg aagaaaacgc
     6361 cagcggcccc atggcccagg tcatcggtcg catggaggcc atcgccgccg ataccgggtg
     6421 ctctatcgtg ttcctgcacc atgccagcaa gggcgcggcc atgatgggcg caggcgacca
     6481 gcagcaggcc agccggggca gctcggtact ggtcgataac atccgctggc agtcctacct
     6541 gtcgagcatg accagcgccg aggccgagga atggggtgtg gacgacgacc agcgccggtt
     6601 cttcgtccgc ttcggtgtga gcaaggccaa ctatggcgca ccgttcgctg atcggtggtt
     6661 caggcggcat gacggcgggg tgctcaagcc cgccgtgctg gagaggcagc gcaagagcaa
     6721 gggggtgccc cgtggtgaag cctaagaaca agcacagcct cagccacgtc cggcacgacc
     6781 cggcgcactg tctggccccc ggcctgttcc gtgccctcaa gcggggcgag cgcaagcgca
     6841 gcaagctgga cgtgacgtat gactacggcg acggcaagcg gatcgagttc agcggcccgg
     6901 agccgctggg cgctgatgat ctgcgcatcc tgcaagggct ggtggccatg gctgggccta
     6961 atggcctagt gcttggcccg gaacccaaga ccgaaggcgg acggcagctc cggctgttcc
     7021 tggaacccaa gtgggaggcc gtcaccgctg aatgccatgt ggtcaaaggt agctatcggg
     7081 cgctggcaaa ggaaatcggg gcagaggtcg atagtggtgg ggcgctcaag cacatacagg
     7141 actgcatcga gcgcctttgg aaggtatcca tcatcgccca gaatggccgc aagcggcagg
     7201 ggtttcggct gctgtcggag tacgccagcg acgaggcgga cgggcgcctg tacgtggccc
     7261 tgaacccctt gatcgcgcag gccgtcatgg gtggcggcca gcatgtgcgc atcagcatgg
     7321 acgaggtgcg ggcgctggac agcgaaaccg cccgcctgct gcaccagcgg ctgtgtggct
     7381 ggatcgaccc cggcaaaacc ggcaaggctt ccatagatac cttgtgcggc tatgtctggc
     7441 cgtcagaggc cagtggttcg accatgcgca agcgccgcca gcgggtgcgc gaggcgttgc
     7501 cggagctggt cgcgctgggc tggacggtaa ccgagttcgc ggcgggcaag tacgacatca
     7561 cccggcccaa ggcggcaggc tgaccccccc cactctattg taaacaagac atttttatct
     7621 tttatattca atggcttatt ttcctgctaa ttggtaatac catgaaaaat accatgctca
     7681 gaaaaggctt aacaatattt tgaaaaattg cctactgagc gctgccgcac agctccatag
     7741 gccgctttcc tggctttgct tccagatgta tgctcttctg ctcccgaacg ccagcaagac
     7801 gtagcccagc gcgtcggcca gcttgcaatt cgcgctaact tacattaatt gcgttgcgct
     7861 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct cgcgagaatt aattcaaaaa
     7921 aagcccgctc attaggcggg ctaattcgag ggcgagacga ggacttccat tgttcattcc
     7981 acggacaaaa acagagaaag gaaacgacag aggccaaaaa gcctcgcttt cagcacctgt
     8041 cgtttccttt cttttcagag ggtattttaa ataaaaacat taagttatga cgaagaagaa
     8101 cggaaacgcc ttaaaccgga aaattttcat aaatagcgaa aacccgcgag gtcgccgccc
     8161 cgtaacctgt cggatcaccg gaaaggaccc gtaaagtgat aatgattatc atctacatat
     8221 cacaacgtgc gtggaggcca tcaaaccacg tcaaataatc aattatgacg caggtatcgt
     8281 attaattgat ctgcatcaac ttaacgtaaa aacaacttca gacaatacaa atcagcgaca
     8341 ctgaatacgg ggcaacctca tgtcctcatc gctaataact tcgtatagca tacattatac
     8401 gaagttatat tcgatg
//
