LOCUS       sequence-357217-        7911 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 GFP promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7911)
  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 7911)
  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: pSL1-GFP
FEATURES             Location/Qualifiers
     source          1..7911
                     /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)"
     terminator      complement(4212..4246)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(4641..5456)
                     /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(4641..5456)
                     /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     5364..5383
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5448..5467
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(5631..6482)
                     /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(6469..7308)
                     /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(7819..7911,1..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 agtaaaggag aagagctttt cacaggagtt gtcccaatcc tcgtggaatt agacggtgat
     3541 gttaatgggc acaagttctc tgtcagtgga gagggtgaag gcgacgcaac atatggcaag
     3601 ctgaccctta aatttatttg caccacgggt aaactacctg ttccatggcc aacactggtc
     3661 actacgttcg ggtatggggt tcagtgcttt gcgcgctacc cagatcacat gaaacagcac
     3721 gactttttca agagtgcaat gcccgaaggc tatgtacagg agagaaccat cttttttaag
     3781 gatgacggca actataagac acgcgccgaa gtgaagttcg agggtgatac ccttgttaat
     3841 agaatcgagt taaagggtat tgactttaag gaagatggaa atattttagg ccacaaactg
     3901 gaatataact ataactccca taatgtgtac attatggccg acaagcaaaa gaacggtatc
     3961 aaggttaact tcaagatcag acacaacatt gaggatggaa gcgttcaact agccgaccat
     4021 taccaacaaa acaccccaat tggcgatggg cctgtgctgt taccagacaa ccattacctg
     4081 tccactcaat ctgccctttc gaaagatccc aacgaaaagc gcgaccacat ggtccttctt
     4141 gagtttgtca cggctgctgg gattacacac ggcatggatg aactatacaa ataaatcctg
     4201 agaccagacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga
     4261 gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcgat atcaaattac
     4321 gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg
     4381 aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct
     4441 tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg
     4501 tttaaatcaa aactggtgaa actcacccag ggattggctg aaacgaaaaa catattctca
     4561 ataaaccctt tagggaaaga gcatttgtca agtcggtctc accgagcggc cgcgtgttac
     4621 aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     4681 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     4741 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     4801 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     4861 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc
     4921 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     4981 ccgttattca ttcgtgattg cgcctgagcg aggcgaaata cgcgatcgct gttaaaagga
     5041 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     5101 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
     5161 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     5221 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     5281 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     5341 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     5401 atgttggaat ttaatcgcgg cctggagcaa gacgtttccc gttgaatatg gctcataaca
     5461 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta
     5521 tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttgttg aataaatcga
     5581 acttttgctg agttgaagga tcagtcatga ccaaaatccc ttaacgtgag tcagcctgcc
     5641 gccttgggcc gggtgatgtc gtacttgccc gccgcgaact cggttaccgt ccagcccagc
     5701 gcgaccagct ccggcaacgc ctcgcgcacc cgctggcggc gcttgcgcat ggtcgaacca
     5761 ctggcctctg acggccagac atagccgcac aaggtatcta tggaagcctt gccggttttg
     5821 ccggggtcga tccagccaca cagccgctgg tgcagcaggc gggcggtttc gctgtccagc
     5881 gcccgcacct cgtccatgct gatgcgcaca tgctggccgc cacccatgac ggcctgcgcg
     5941 atcaaggggt tcagggccac gtacaggcgc ccgtccgcct cgtcgctggc gtactccgac
     6001 agcagccgaa acccctgccg cttgcggcca ttctgggcga tgatggatac cttccaaagg
     6061 cgctcgatgc agtcctgtat gtgcttgagc gccccaccac tatcgacctc tgccccgatt
     6121 tcctttgcca gcgcccgata gctacctttg accaccatgg catcagcggt gacggcctcc
     6181 cacttgggtt ccaggaacag ccggagctgc cgtccgcctt cggtcttggg ttccgggcca
     6241 agcactaggc cattaggccc agccatggcc accagccctt gcaggatgcg cagatcatca
     6301 gcgcccagcg gctccgggcc gctgaactcg atccgcttgc cgtcgccgta gtcatacgtc
     6361 acgtccagct tgctgcgctt gcgctcgccc cgcttgaggg cacggaacag gccgggggcc
     6421 agacagtgcg ccgggtcgtg ccggacgtgg ctgaggctgt gcttgttctt aggcttcacc
     6481 acggggcacc cccttgctct tgcgctgcct ctccagcacg gcgggcttga gcaccccgcc
     6541 gtcatgccgc ctgaaccacc gatcagcgaa cggtgcgcca tagttggcct tgctcacacc
     6601 gaagcggacg aagaaccggc gctggtcgtc gtccacaccc cattcctcgg cctcggcgct
     6661 ggtcatgctc gacaggtagg actgccagcg gatgttatcg accagtaccg agctgccccg
     6721 gctggcctgc tgctggtcgc ctgcgcccat catggccgcg cccttgctgg catggtgcag
     6781 gaacacgata gagcacccgg tatcggcggc gatggcctcc atgcgaccga tgacctgggc
     6841 catggggccg ctggcgtttt cttcctcgat gtggaaccgg cgcagcgtgt ccagcaccat
     6901 caggcggcgg ccctcggcgg cgcgcttgag gccgtcgaac cactccgggg ccatgatgtt
     6961 gggcaggctg ccgatcagcg gctggatcag caggccgtca gccacggctt gccgttcctc
     7021 ggcgctgagg tgcgccccaa gggcgtgcag gcggtgatga atggcggtgg gcgggtcttc
     7081 ggcgggcagg tagatcaccg ggccggtggg cagttcgccc acctccagca gatccggccc
     7141 gcctgcaatc tgtgcggcca gttgcagggc cagcatggat ttaccggcac caccgggcga
     7201 caccagcgcc ccgaccgtac cggccaccat gttgggcaaa acgtagtcca gcggtggcgg
     7261 cgctgctgcg aacgcctcca gaatattgat aggcttatgg gtagccattg attgcctcct
     7321 ttgcaggcag ttggtggtta ggcgctggcg gggtcactac ccccgccctg cgccgctctg
     7381 agttcttcca ggcactcgcg cagcgcctcg tattcgtcgt cggtcagcca gaacttgcgc
     7441 tgacgcatcc ctttggcctt catgcgctcg gcatatcgcg cttggcgtac agcgtcaggg
     7501 ctggccagca ggtcgccggt ctgcttgtcc ttttggtctt tcatatcagt caccgagaaa
     7561 cttgccgggg ccgaaaggct tgtcttcgcg gaacaaggac aaggtgcagc cgtcaaggtt
     7621 aaggctggcc atatcagcga ctgaaaagcg gccagcctcg gccttgtttg acgtataacc
     7681 aaagccaccg ggcaaccaat agcccttgtc acttttgatc aggtagaccg accctgaagc
     7741 gcttttttcg tattccataa aacccccttc tgtgcgtgag tactcatagt ataacaggcg
     7801 tgagtaccaa cgcaagcact acatgctgaa atctggcccg cccctgtcca tgcctcgctg
     7861 gcggggtgcc ggtgcccgtg ccagctcggc ccgcgcaagc tggacgctgg g
//
