LOCUS       sequence-260869-        7374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RSF1010-based broad host-range plasmid with additional RK2 bom site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7374)
  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 7374)
  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: pAM5404
FEATURES             Location/Qualifiers
     source          1..7374
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(121..912,913..981))
                     /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     744..763
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(982..1086)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1154..1172
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(1647..2498)
                     /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(2485..3324)
                     /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(3835..4806)
                     /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            6045..6132
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      6474..6868
                     /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(7031..7074)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     oriT            7170..7279
                     /note="incP origin of transfer"
ORIGIN
        1 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
       61 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
      121 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
      181 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      241 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
      301 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      361 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      421 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      481 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      541 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      601 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      661 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      721 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      781 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      841 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      901 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      961 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     1021 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     1081 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     1141 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     1201 cggtgaaaac ctctgacaca tgcagcgacg tcgggggccc ccggggggat tctagaacaa
     1261 tggaagtcct cgtctcgccc tcgaattagc ccgcttaatg agcgggcttt ttttgaatta
     1321 attctcgcga gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     1381 attaatgtaa gttagcgcga attgcaagct ggccgacgcg ctgggctacg tcttgctggc
     1441 gttcgggagc agaagagcat acatctggaa gcaaagccag gaaagcggcc tatggagctg
     1501 tgcggcagcg ctcagtaggc aatttttcaa aatattgtta agccttttct gagcatggta
     1561 tttttcatgg tattaccaat tagcaggaaa ataagccatt gaatataaaa gataaaaatg
     1621 tcttgtttac aatagagtgg ggggggtcag cctgccgcct tgggccgggt gatgtcgtac
     1681 ttgcccgccg cgaactcggt taccgtccag cccagcgcga ccagctccgg caacgcctcg
     1741 cgcacccgct ggcggcgctt gcgcatggtc gaaccactgg cctctgacgg ccagacatag
     1801 ccgcacaagg tatctatgga agccttgccg gttttgccgg ggtcgatcca gccacacagc
     1861 cgctggtgca gcaggcgggc ggtttcgctg tccagcgccc gcacctcgtc catgctgatg
     1921 cgcacatgct ggccgccacc catgacggcc tgcgcgatca aggggttcag ggccacgtac
     1981 aggcgcccgt ccgcctcgtc gctggcgtac tccgacagca gccgaaaccc ctgccgcttg
     2041 cggccattct gggcgatgat ggataccttc caaaggcgct cgatgcagtc ctgtatgtgc
     2101 ttgagcgccc caccactatc gacctctgcc ccgatttcct ttgccagcgc ccgatagcta
     2161 cctttgacca ccatggcatc agcggtgacg gcctcccact tgggttccag gaacagccgg
     2221 agctgccgtc cgccttcggt cttgggttcc gggccaagca ctaggccatt aggcccagcc
     2281 atggccacca gcccttgcag gatgcgcaga tcatcagcgc ccagcggctc cgggccgctg
     2341 aactcgatcc gcttgccgtc gccgtagtca tacgtcacgt ccagcttgct gcgcttgcgc
     2401 tcgccccgct tgagggcacg gaacaggccg ggggccagac agtgcgccgg gtcgtgtcgg
     2461 acgtggctga ggctgtgctt gttcttaggc ttcaccacgg ggcaccccct tgctcttgcg
     2521 ctgcctctcc agcacggcgg gcttgagcac cccgccgtca tgccgcctga accaccgatc
     2581 agcgaacggt gcgccatagt tggccttgct cacaccgaag cggacgaaga accggcgctg
     2641 gtcgtcgtcc acaccccatt cctcggcctc ggcgctggtc atgctcgaca ggtaggactg
     2701 ccagcggatg ttatcgacca gtaccgagct gccccggctg gcctgctgct ggtcgcctgc
     2761 gcccatcatg gccgcgccct tgctggcatg gtgcaggaac acgatagagc acccggtatc
     2821 ggcggcgatg gcctccatgc gaccgatgac ctgggccatg gggccgctgg cgttttcttc
     2881 ctcgatgtgg aaccggcgca gcgtgtccag caccatcagg cggcggccct cggcggcgcg
     2941 cttgaggccg tcgaaccact ccggggccat gatgttgggc aggctgccga tcagcggctg
     3001 gatcagcagg ccgtcagcca cggcttgccg ttcctcggcg ctgaggtgcg ccccaagggc
     3061 gtgcaggcgg tgatgaatgg cggtgggcgg gtcttcggcg ggcaggtaga tcaccgggcc
     3121 ggtgggcagt tcgcccacct ccagcagatc cggcccgcct gcaatctgtg cggccagttg
     3181 cagggccagc atggatttac cggcaccacc gggcgacacc agcgccccga ccgtaccggc
     3241 caccatgttg ggcaaaacgt agtccagcgg tggcggcgct gctgcgaacg cctccagaat
     3301 attgataggc ttatgggtag ccattgattg cctcctttgc aggcagttgg tggttaggcg
     3361 ctggcggggt cactaccccc gccctgcgcc gctctgagtt cttccaggca ctcgcgcagc
     3421 gcctcgtatt cgtcgtcggt cagccagaac ttgcgctgac gcatcccttt ggccttcatg
     3481 cgctcggcat atcgcgcttg gcgtacagcg tcagggctgg ccagcaggtc gccggtctgc
     3541 ttgtcctttt ggtctttcat atcagtcacc gagaaacttg ccggggccga aaggcttgtc
     3601 ttcgcggaac aaggacaagg tgcagccgtc aaggttaagg ctggccatat cagcgactga
     3661 aaagcggcca gcctcggcct tgtttgacgt ataaccaaag ccaccgggca accaatagcc
     3721 cttgtcactt ttgatcaggt agaccgaccc tgaagcgctt ttttcgtatt ccataaaacc
     3781 cccttctgtg cgtgagtact catagtataa caggcgtgag taccaacgca agcactacat
     3841 gctgaaatct ggcccgcccc tgtccatgcc tcgctggcgg ggtgccggtg cccgtgccag
     3901 ctcggcccgc gcaagctgga cgctgggcag acccatgacc ttgctgacgg tgcgctcgat
     3961 gtaatccgct tcgtggccgg gcttgcgctc tgccagcgct gggctggcct cggccatggc
     4021 cttgccgatt tcctcggcac tgcggccccg gctggccagc ttctgcgcgg cgataaagtc
     4081 gcacttgctg aggtcatcac cgaagcgctt gaccagcccg gccatctcgc tgcggtactc
     4141 gtccagcgcc gtgcgccggt ggcggctaag ctgccgctcg gacagttcga ggctggccag
     4201 cctgcgggcc ttctcctgct gccgctgggc ctgctcgatc tgctggccag cctgctgcac
     4261 cagcgccggg ccagcggtgg cggtcttgcc cttggattca cgcagcagca cccacggctg
     4321 ataaccggcg cgggtggtgt gcttgtcctt gcggttggtg aagcccgcca agcggccata
     4381 gtggcggctg tcggcgctgg ccgggtcggc gtcgtactcg ctggccagcg tccgggcaat
     4441 ctgcccccga agttcaccgc ctgcggcgtc ggccaccttg acccatgcct gatagttctt
     4501 cgggctggtt tccactacca gggcaggctc ccggccctcg gctttcatgt catccaggtc
     4561 aaactcgctg aggtcgtcca ccagcaccag accatgccgc tcctgctcgg cgggcctgat
     4621 atacacgtca ttgccctggg cattcatccg cttgagccat ggcgtgttct ggagcacttc
     4681 ggcggctgac cattcccggt tcatcatctg gccggtggtg gcgtccctga cgccgatatc
     4741 gaagcgctca cagcccatgg ccttgagctg tcggcctatg gcctgcaaag tcctgtcgtt
     4801 cttcatcggg ccaccaagcg cagccagatc gagccgtcct cggttgtcag tggcgtcagg
     4861 tcgagcaaga gcaacgatgc gatcagcagc accaccgtag gcatcatgga agccagcatc
     4921 acggttagcc atagcttcca gtgccacccc cgcgacgcgc tccgggcgct ctgcgcggcg
     4981 ctgctcacct cggcggctac ctcccgcaac tctttggcca gctccaccca tgccgcccct
     5041 gtctggcgct gggctttcag ccactccgcc gcctgcgcct cgctggcctg ctgggtctgg
     5101 ctcatgacct gccgggcttc gtcggccagt gtcgccatgc tctgggccag cggttcgatc
     5161 tgctccgcta actcgttgat gcctctggat ttcttcactc tgtcgattgc gttcatggtc
     5221 tattgcctcc cggtattcct gtaagtcgat gatctgggcg ttggcggtgt cgatgttcag
     5281 ggccacgtct gcccggtcgg tgcggatgcc ccggccttcc atctccacca cgttcggccc
     5341 caggtgaaca ccgggcaggc gctcgatgcc ctgcgcctca agtgttctgt ggtcaatgcg
     5401 ggcgtcgtgg cctgcccgct ctaatgcccg gttggcatgg tcggcccatg cctcgcgggt
     5461 ctgctcaagc catgccttgg gcttgagcgc ttcggtcttc tgtgccccgc ccttctccgg
     5521 ggtcttgccg ttgtaccgct tgaaccactg agcggcgggc cgctcgatgc cgtcattgat
     5581 ccgctcggag atcatcaggt ggcagtgcgg gttctcgccg ccaccggcat ggatggccag
     5641 cgtatacggc aggcgctcgg caccggtcag gtgctgggcg aactcggacg ccagcgcctt
     5701 ctgctggtcg agggtcagct cgaccggcag ggcaaattcg acctccttga acagccgccc
     5761 attggcgcgt tcatacaggt cggcagcatc ccagtagtcg gcgggccgct cgacgaactc
     5821 cggcatgtgc ccggattcgg cgtgcaagac ttcatccatg tcgcgggcat acttgccttc
     5881 gcgctggatg aagtcggcct tggccctggc cgattggccg cccgacctgc tgccggtttt
     5941 cgccgtaagg tgataaatcg ccatgctgcc tcgctgttgc ttttgctttt cggctccatg
     6001 caatggccct cggagagcgc accgcccgaa gggtggccgt taggccagtt tctcgaagag
     6061 aaaccggtaa gtgcgccctc ccctacaaag tagggtcggg attgccgccg ctgtgcctcc
     6121 atgatagcct acgagacagc acattaacaa tggggtgtca agatggttaa ggggagcaac
     6181 aaggcggcgg atcggctggc caagctcgaa gaacaacgag cgcgaatcaa tgccgaaatt
     6241 cagcgggtgc gggcaaggga acagcagcaa gagcgcaaga acgaaacaag gcgcaaggtg
     6301 ctggtggggg ccatgatttt ggccaaggtg aacagcagcg agtggccgga ggatcggctc
     6361 atggcggcaa tggatgcgta ccttgaacgc gaccacgacc gcgccttgtt cggtctgccg
     6421 ccacgccaga aggatgagcc gggctgaatg atcgaccgag acaggccctg cggggctgca
     6481 cacgcgcccc cacccttcgg gtagggggaa aggccgctaa agcggctaaa agcgctccag
     6541 cgtatttctg cggggtttgg tgtggggttt agcgggcttt gcccgccttt ccccctgccg
     6601 cgcagcggtg gggcggtgtg tagcctagcg cagcgaatag accagctatc cggcctctgg
     6661 ccgggcatat tgggcaaggg cagcagcgcc ccacaagggc gctgataacc gcgcctagtg
     6721 gattattctt agataatcat ggatggattt ttccaacacc ccgccagccc ccgcccctgc
     6781 tgggtttgca ggtttggggg cgtgacagtt attgcagggg ttcgtgacag ttattgcagg
     6841 ggggcgtgac agttattgca ggggttcgtg acagttagta cgggagtgac gggcactggc
     6901 tggcaatgtc tagcaacggc aggcatttcg gctgagggta aaagaacttt ccgctaagcg
     6961 atagactgta tgtaaacaca gtattgcaag gacgcggaac atgcctcatg tggcggccct
     7021 cgcgagaatt aattcagata aaaaaaatcc ttagctttcg ctaaggatga tttctagcga
     7081 tgaccctgct gattggttcg ctgaccgtag actggttggc ttggtttcat cagccatccg
     7141 cttgccctca tctgttacgc cggcggtagc cggccagcct cgcagagcag gattcccgtt
     7201 gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa cacccgctcg cgggtgggcc
     7261 tacttcacct atcctgcccg gctgacgccg ttggatacac caaggaaagt ctacacgaac
     7321 cctttggcaa aatcctgtat atcgtgcgac atcgcgcgcg cgcgcgcgga cgtc
//
