LOCUS       sequence-357215-        7616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Kan resistant plasmid derived from pBTK402, a broad-host-range 
            plasmid previously shown to replicate in multiple species of bee gut
            bacteria. It encodes a low level RFP promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7616)
  AUTHORS   Li Y, Leonard SP, Powell JE, Moran NA
  TITLE     Species divergence in gut-restricted bacteria of social bees.
  JOURNAL   Proc Natl Acad Sci U S A. 2022 May 3;119(18):e2115013119. doi: 
            10.1073/pnas.2115013119. Epub 2022 Apr 25.
  PUBMED    35467987
REFERENCE   2  (bases 1 to 7616)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2022-05-3"; volume: "119(18):e2115013119. 
            doi"; pages: " 10.1073/pnas.2115013119. Epub 2022 Apr 25"
COMMENT     Sequence Label: pSL-1
FEATURES             Location/Qualifiers
     source          1..7616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            2118..2205
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      2547..2941
                     /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)"
     CDS             3478..4155
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      4175..4246
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4262..4289
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4346..5161)
                     /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(4346..5161)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5069..5088
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5153..5172
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(5336..6187)
                     /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(6174..7013)
                     /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(join(7524..7616,1..879))
                     /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"
ORIGIN
        1 cagacccatg accttgctga cggtgcgctc gatgtaatcc gcttcgtggc cgggcttgcg
       61 ctctgccagc gctgggctgg cctcggccat ggccttgccg atttcctcgg cactgcggcc
      121 ccggctggcc agcttctgcg cggcgataaa gtcgcacttg ctgaggtcat caccgaagcg
      181 cttgaccagc ccggccatct cgctgcggta ctcgtccagc gccgtgcgcc ggtggcggct
      241 aagctgccgc tcgggcagtt cgaggctggc cagcctgcgg gccttctcct gctgccgctg
      301 ggcctgctcg atctgctggc cagcctgctg caccagcgcc gggccagcgg tggcggtctt
      361 gcccttggat tcacgcagca gcacccacgg ctgataaccg gcgcgggtgg tgtgcttgtc
      421 cttgcggttg gtgaagcccg ccaagcggcc atagtggcgg ctgtcggcgc tggccgggtc
      481 ggcgtcgtac tcgctggcca gcgtccgggc aatctgcccc cgaagttcac cgcctgcggc
      541 gtcggccacc ttgacccatg cctgatagtt cttcgggctg gtttccacta ccagggcagg
      601 ctcccggccc tcggctttca tgtcatccag gtcaaactcg ctgaggtcgt ccaccagcac
      661 cagaccatgc cgctcctgct cggcgggcct gatatacacg tcattgccct gggcattcat
      721 ccgcttgagc catggcgtgt tctggagcac ttcggcggct gaccattccc ggttcatcat
      781 ctggccggtg gtggcgtccc tgacgccgat atcgaagcgc tcacagccca tggccttgag
      841 ctgtcggcct atggcctgca aagtcctgtc gttcttcatc gggccaccaa gcgcagccag
      901 atcgagccgt cctcggttgt cagtggcgtc aggtcgagca agagcaacga tgcgatcagc
      961 agcaccaccg taggcatcat ggaagccagc atcacggtta gccatagctt ccagtgccac
     1021 ccccgcgacg cgctccgggc gctctgcgcg gcgctgctca cctcggcggc tacctcccgc
     1081 aactctttgg ccagctccac ccatgccgcc cctgtctggc gctgggcttt cagccactcc
     1141 gccgcctgcg cctcgctggc ctgctgggtc tggctcatga cctgccgggc ttcgtcggcc
     1201 agtgtcgcca tgctctgggc cagcggttcg atctgctccg ctaactcgtt gatgcctctg
     1261 gatttcttca ctctgtcgat tgcgttcatg gtctattgcc tcccggtatt cctgtaagtc
     1321 gatgatctgg gcgttggcgg tgtcgatgtt cagggccacg tctgcccggt cggtgcggat
     1381 gccccggcct tccatctcca ccacgttcgg ccccaggtga acaccgggca ggcgctcgat
     1441 gccctgcgcc tcaagtgttc tgtggtcaat gcgggcgtcg tggccagccc gctctaatgc
     1501 ccggttggca tggtcggccc atgcctcgcg ggtctgctca agccatgcct tgggcttgag
     1561 cgcttcggtc ttctgtgccc cgcccttctc cggggtcttg ccgttgtacc gcttgaacca
     1621 ctgagcggcg ggccgctcga tgccgtcatt gatccgctcg gagatcatca ggtggcagtg
     1681 cgggttctcg ccgccaccgg catggatggc cagcgtatac ggcaggcgct cggcaccggt
     1741 caggtgctgg gcgaactcgg acgccagcgc cttctgctgg tcgagggtca gctcgaccgg
     1801 cagggcaaat tcgacctcct tgaacagccg cccattggcg cgttcataca ggtcggcagc
     1861 atcccagtag tcggcgggcc gctcgacgaa ctccggcatg tgcccggatt cggcgtgcaa
     1921 gacttcatcc atgtcgcggg catacttgcc ttcgcgctgg atgtagtcgg ccttggccct
     1981 ggccgattgg ccgcccgacc tgctgccggt tttcgccgta aggtgataaa tcgccatgct
     2041 gcctcgctgt tgcttttgct tttcggctcc atgcaatggc cctcggagag cgcaccgccc
     2101 gaagggtggc cgttaggcca gtttctcgaa gagaaaccgg taagtgcgcc ctcccctaca
     2161 aagtagggtc gggattgccg ccgctgtgcc tccatgatag cctacgagac agcacattaa
     2221 caatggggtg tcaagatggt taaggggagc aacaaggcgg cggatcggct ggccaagctc
     2281 gaagaacaac gagcgcgaat caatgccgaa attcagcggg tgcgggcaag ggaacagcag
     2341 caagagcgca agaacgaaac aaggcgcaag gtgctggtgg gggccatgat tttggccaag
     2401 gtgaacagca gcgagtggcc ggaggatcgg ctcatggcgg caatggatgc gtaccttgaa
     2461 cgcgaccacg accgcgcctt gttcggtctg ccgccacgcc agaaggatga gccgggctga
     2521 atgatcgacc gagacaggcc ctgcggggct gcacacgcgc ccccaccctt cgggtagggg
     2581 gaaaggccgc taaagcggct aaaagcgctc cagcgtattt ctgcggggtt tggtgtgggg
     2641 tttagcgggc tttgcccgcc tttccccctg ccgcgcagcg gtggggcggt gtgtagccta
     2701 gcgcagcgaa tagaccagct atccggcctc tggccgggca tattgggcaa gggcagcagc
     2761 gccccacaag ggcgctgata accgcgccta gtggattatt cttagataat catggatgga
     2821 tttttccaac accccgccag cccccgcccc tgctgggttt gcaggtttgg gggcgtgaca
     2881 gttattgcag gggttcgtga cagttattgc aggggggcgt gacagttatt gcaggggttc
     2941 gtgacagtta gtacgggagt gacgggcact ggctggcaat gtctagcaac ggcaggcatt
     3001 tcggctgagg gtaaaagaac tttccgctaa gcgatagact gtatgtaaac acagtattgc
     3061 aaggacgcgg aacatgcctc atgtggcggc caggacggcc agccgggatc gggatactgg
     3121 tcgttaccag agccaccgac ccgagcaaac ccttctctat cagatcgttg acgagtatta
     3181 cccggcattc gctgcgctta tggcagagca gggaaaggaa ttgccgggct atgtgcaacg
     3241 ggaatttgaa gaatttctcc aatgcgggcg gctggagcat ggctttctac gggttcgctg
     3301 cgagtcttgc cacgccgagc acctggtcgc tttcagaaat caatctaaag tatatatgag
     3361 taaacttggt ctgacaggcc ccttgagacc agctttggca gtttattctt gacatgtagt
     3421 gagggggctg gtataatcac atagtactgt tatacagaaa cagaggagat attacatatg
     3481 gcttcctccg aagacgttat caaagagttc atgcgtttca aagttcgtat ggaaggttcc
     3541 gttaacggtc acgagttcga aatcgaaggt gaaggtgaag gtcgtccgta cgaaggtacc
     3601 cagaccgcta aactgaaagt taccaaaggt ggtccgctgc cgttcgcttg ggacatcctg
     3661 tccccgcagt tccagtacgg ttccaaagct tacgttaaac acccggctga catcccggac
     3721 tacctgaaac tgtccttccc ggaaggtttc aaatgggaac gtgttatgaa cttcgaagac
     3781 ggtggtgttg ttaccgttac ccaggactcc tccctgcaag acggtgagtt catctacaaa
     3841 gttaaactgc gtggtaccaa cttcccgtcc gacggtccgg ttatgcagaa aaaaaccatg
     3901 ggttgggaag cttccaccga acgtatgtac ccggaagacg gtgctctgaa aggtgaaatc
     3961 aaaatgcgtc tgaaactgaa agacggtggt cactacgacg ctgaagttaa aaccacctac
     4021 atggctaaaa aaccggttca gctgccgggt gcttacaaaa ccgacatcaa actggacatc
     4081 acctcccaca acgaagacta caccatcgtt gaacagtacg aacgtgctga aggtcgtcac
     4141 tccaccggcg cttaatataa ggatctccag gcatcaaata aaacgaaagg ctcagtcgaa
     4201 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact agagtcacac
     4261 tggctcacct tcgggtgggc ctttctgcgt ttataagtcg gtctcaccga gcggccgcgt
     4321 gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca
     4381 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     4441 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     4501 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     4561 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt
     4621 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     4681 ccaaaccgtt attcattcgt gattgcgcct gagcgaggcg aaatacgcga tcgctgttaa
     4741 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     4801 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga
     4861 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     4921 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     4981 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     5041 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     5101 catccatgtt ggaatttaat cgcggcctgg agcaagacgt ttcccgttga atatggctca
     5161 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat
     5221 ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt tgttgaataa
     5281 atcgaacttt tgctgagttg aaggatcagt catgaccaaa atcccttaac gtgagtcagc
     5341 ctgccgcctt gggccgggtg atgtcgtact tgcccgccgc gaactcggtt accgtccagc
     5401 ccagcgcgac cagctccggc aacgcctcgc gcacccgctg gcggcgcttg cgcatggtcg
     5461 aaccactggc ctctgacggc cagacatagc cgcacaaggt atctatggaa gccttgccgg
     5521 ttttgccggg gtcgatccag ccacacagcc gctggtgcag caggcgggcg gtttcgctgt
     5581 ccagcgcccg cacctcgtcc atgctgatgc gcacatgctg gccgccaccc atgacggcct
     5641 gcgcgatcaa ggggttcagg gccacgtaca ggcgcccgtc cgcctcgtcg ctggcgtact
     5701 ccgacagcag ccgaaacccc tgccgcttgc ggccattctg ggcgatgatg gataccttcc
     5761 aaaggcgctc gatgcagtcc tgtatgtgct tgagcgcccc accactatcg acctctgccc
     5821 cgatttcctt tgccagcgcc cgatagctac ctttgaccac catggcatca gcggtgacgg
     5881 cctcccactt gggttccagg aacagccgga gctgccgtcc gccttcggtc ttgggttccg
     5941 ggccaagcac taggccatta ggcccagcca tggccaccag cccttgcagg atgcgcagat
     6001 catcagcgcc cagcggctcc gggccgctga actcgatccg cttgccgtcg ccgtagtcat
     6061 acgtcacgtc cagcttgctg cgcttgcgct cgccccgctt gagggcacgg aacaggccgg
     6121 gggccagaca gtgcgccggg tcgtgccgga cgtggctgag gctgtgcttg ttcttaggct
     6181 tcaccacggg gcaccccctt gctcttgcgc tgcctctcca gcacggcggg cttgagcacc
     6241 ccgccgtcat gccgcctgaa ccaccgatca gcgaacggtg cgccatagtt ggccttgctc
     6301 acaccgaagc ggacgaagaa ccggcgctgg tcgtcgtcca caccccattc ctcggcctcg
     6361 gcgctggtca tgctcgacag gtaggactgc cagcggatgt tatcgaccag taccgagctg
     6421 ccccggctgg cctgctgctg gtcgcctgcg cccatcatgg ccgcgccctt gctggcatgg
     6481 tgcaggaaca cgatagagca cccggtatcg gcggcgatgg cctccatgcg accgatgacc
     6541 tgggccatgg ggccgctggc gttttcttcc tcgatgtgga accggcgcag cgtgtccagc
     6601 accatcaggc ggcggccctc ggcggcgcgc ttgaggccgt cgaaccactc cggggccatg
     6661 atgttgggca ggctgccgat cagcggctgg atcagcaggc cgtcagccac ggcttgccgt
     6721 tcctcggcgc tgaggtgcgc cccaagggcg tgcaggcggt gatgaatggc ggtgggcggg
     6781 tcttcggcgg gcaggtagat caccgggccg gtgggcagtt cgcccacctc cagcagatcc
     6841 ggcccgcctg caatctgtgc ggccagttgc agggccagca tggatttacc ggcaccaccg
     6901 ggcgacacca gcgccccgac cgtaccggcc accatgttgg gcaaaacgta gtccagcggt
     6961 ggcggcgctg ctgcgaacgc ctccagaata ttgataggct tatgggtagc cattgattgc
     7021 ctcctttgca ggcagttggt ggttaggcgc tggcggggtc actacccccg ccctgcgccg
     7081 ctctgagttc ttccaggcac tcgcgcagcg cctcgtattc gtcgtcggtc agccagaact
     7141 tgcgctgacg catccctttg gccttcatgc gctcggcata tcgcgcttgg cgtacagcgt
     7201 cagggctggc cagcaggtcg ccggtctgct tgtccttttg gtctttcata tcagtcaccg
     7261 agaaacttgc cggggccgaa aggcttgtct tcgcggaaca aggacaaggt gcagccgtca
     7321 aggttaaggc tggccatatc agcgactgaa aagcggccag cctcggcctt gtttgacgta
     7381 taaccaaagc caccgggcaa ccaatagccc ttgtcacttt tgatcaggta gaccgaccct
     7441 gaagcgcttt tttcgtattc cataaaaccc ccttctgtgc gtgagtactc atagtataac
     7501 aggcgtgagt accaacgcaa gcactacatg ctgaaatctg gcccgcccct gtccatgcct
     7561 cgctggcggg gtgccggtgc ccgtgccagc tcggcccgcg caagctggac gctggg
//
