LOCUS       sequence-314921-        8618 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-Host range expression vector, RSF1010 origin, TetR repressor, 
            PTetA promoter, mRFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8618)
  AUTHORS   Schuster LA, Reisch CR
  TITLE     A plasmid toolbox for controlled gene expression across the 
            Proteobacteria.
  JOURNAL   Nucleic Acids Res. 2021 Jun 14. pii: 6298615. doi: 
            10.1093/nar/gkab496.
  PUBMED    34125913
REFERENCE   2  (bases 1 to 8618)
  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: "Nucleic 
            Acids Res. 2021 Jun 14. pii: 6298615. doi: 10.1093/nar/gkab496."
COMMENT     Sequence Label: pFTG5
FEATURES             Location/Qualifiers
     source          1..8618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      330..724
                     /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            1066..1153
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             2392..3363
                     /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             3874..4713
                     /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             4700..5551
                     /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"
     primer_bind     5975..5997
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5989..6006
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5990..6006
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(6111..6737)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     6642..6661
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        6753..6807
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    6759..6777
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    6789..6807
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             6829..6840
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             6847..7524
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     promoter        join(7801..7806,7807..7823,7824..7829)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             8018..8551
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 ggtggcactt ttcggggatc ttcaaattcc cgttgcacat agcccggcaa ttcctttccc
       61 tgctctgcca taagcgcagc gaatgccggg taatactcgt caacgatctg atagagaagg
      121 gtttgctcgg gtcggtggct ctggtaacga ccagtatccc gatcccggct ggccgtcctg
      181 gccgccacat gaggcatgtt ccgcgtcctt gcaatactgt gtttacatac agtctatcgc
      241 ttagcggaaa gttcttttac cctcagccga aatgcctgcc gttgctagac attgccagcc
      301 agtgcccgtc actcccgtac taactgtcac gaacccctgc aataactgtc acgcccccct
      361 gcaataactg tcacgaaccc ctgcaataac tgtcacgccc ccaaacctgc aaacccagca
      421 ggggcggggg ctggcggggt gttggaaaaa tccatccatg attatctaag aataatccac
      481 taggcgcggt tatcagcgcc cttgtggtgc gctgctgccc ttgcccaata tgcccggcca
      541 gaggccggat agctggtcta ttcgctgcgc taggctacac accgccccac cgctgcgcgg
      601 cagggggaaa ggcgggcaaa gcccgctaaa ccccacacca aaccccgcag aaatacgctg
      661 gagcgctttt agccgcttta gcggcctttc cccctacccg aagggtgggg gcgcgtgtgc
      721 agccccgcag ggcctgtctc ggtcgatcat tcagcccggc tcatccttct ggcgtggcgg
      781 cagaccgaac aaggcgcggt cgtggtcgcg ttcaaggtac gcatccattg ccgccatgag
      841 ccgatcctcc ggccactcgc tgctgttcac cttggccaaa atcatggccc ccaccagcac
      901 cttgcgcctt gtttcgttct tgcgctcttg ctgctgttcc cttgcccgca cccgctgaat
      961 ttcggcattg attcgcgctc gttgttcttc gagcttggcc agccgatccg ccgccttgtt
     1021 gctcccctta accatcttga caccccattg ttaatgtgct gtctcgtagg ctatcatgga
     1081 ggcacagcgg cggcaatccc gaccctactt tgtaggggag ggcgcactta ccggtttctc
     1141 ttcgagaaac tggcctaacg gccacccttc gggcggtgcg ctctccgagg gccattgcat
     1201 ggagccgaaa agcaaaagca acagcgaggc agcatggcga tttatcacct tacggcgaaa
     1261 accggcagca ggtcgggcgg ccaatcggcc agggccaagg ccgactacat ccagcgcgaa
     1321 ggcaagtatg cccgcgacat ggatgaagtc ttgcacgccg aatccgggca catgccggag
     1381 ttcgtcgagc ggcccgccga ctactgggat gctgccgacc tgtatgaacg cgccaatggg
     1441 cggctgttca aggaggtcga atttgccctg ccggtcgagc tgaccctcga ccagcagaag
     1501 gcgctggcgt ccgagttcgc ccagcacctg accggtgccg agcgcctgcc gtatacgctg
     1561 gccatccatg ccggtggcgg cgagaacccg cactgccacc tgatgatctc cgagcggatc
     1621 aatgacggca tcgagcggcc cgccgctcag tggttcaagc ggtacaacgg caagaccccg
     1681 gagaagggcg gggcacagaa gaccgaagcg ctcaagccca aggcatggct tgagcagacc
     1741 cgcgaggcat gggccgacca tgccaaccgg gcattagagc gggctggcca cgacgcccgc
     1801 attgaccaca gaacacttga ggcgcagggc atcgagcgcc tgcccggtgt tcacctgggg
     1861 ccgaacgtgg tggagatgga aggccggggc atccgcaccg accgggcaga cgtggccctg
     1921 aacatcgaca ccgccaacgc ccagatcatc gacttacagg aataccggga ggcaatagac
     1981 catgaacgca atcgacagag tgaagaaatc cagaggcatc aacgagttag cggagcagat
     2041 cgaaccgctg gcccagagca tggcgacact ggccgacgaa gcccggcagg tcatgagcca
     2101 gacccagcag gccagcgagg cgcaggcggc ggagtggctg aaagcccagc gccagacagg
     2161 ggcggcatgg gtggagctgg ccaaagagtt gcgggaggta gccgccgagg tgagcagcgc
     2221 cgcgcatagc gcccggagcg cgtcgcgggg gtggcactgg aagctatggc taaccgtgat
     2281 gctggcttcc atgatgccta cggtggtgct gctgatcgca tcgttgctct tgctcgacct
     2341 gacgccactg acaaccgagg acggctcgat ctggctgcgc ttggtggccc gatgaagaac
     2401 gacaggactt tgcaggccat aggccgacag ctcaaggcca tgggctgtga gcgcttcgat
     2461 atcggcgtca gggacgccac caccggccag atgatgaacc gggaatggtc agccgccgaa
     2521 gtgctccaga acacgccatg gctcaagcgg atgaatgccc agggcaatga cgtgtatatc
     2581 aggcccgccg agcaggagcg gcatggtctg gtgctggtgg acgacctcag cgagtttgac
     2641 ctggatgaca tgaaagccga gggccgggag cctgccctgg tagtggaaac cagcccgaag
     2701 aactatcagg catgggtcaa ggtggccgac gccgcaggcg gtgaacttcg ggggcagatt
     2761 gcccggacgc tggccagcga gtacgacgcc gacccggcca gcgccgacag ccgccactat
     2821 ggccgcttgg cgggcttcac caaccgcaag gacaagcaca ccacccgcgc cggttatcag
     2881 ccgtgggtgc tgctgcgtga atccaagggc aagaccgcca ccgctggccc ggcgctggtg
     2941 cagcaggctg gccagcagat cgagcaggcc cagcggcagc aggagaaggc ccgcaggctg
     3001 gccagcctcg aactgcccga gcggcagctt agccgccacc ggcgcacggc gctggacgag
     3061 taccgcagcg agatggccgg gctggtcaag cgcttcggtg atgacctcag caagtgcgac
     3121 tttatcgccg cgcagaagct ggccagccgg ggccgcagtg ccgaggaaat cggcaaggcc
     3181 atggccgagg ccagcccagc gctggcagag cgcaagcccg gccacgaagc ggattacatc
     3241 gagcgcaccg tcagcaaggt catgggtctg cccagcgtcc agcttgcgcg ggccgagctg
     3301 gcacgggcac cggcaccccg ccagcgaggc atggacaggg gcgggccaga tttcagcatg
     3361 tagtgcttgc gttggtactc acgcctgtta tactatgagt actcacgcac agaagggggt
     3421 tttatggaat acgaaaaaag cgcttcaggg tcggtctacc tgatcaaaag tgacaagggc
     3481 tattggttgc ccggtggctt tggttatacg tcaaacaagg ccgaggctgg ccgcttttca
     3541 gtcgctgata tggccagcct taaccttgac ggctgcacct tgtccttgtt ccgcgaagac
     3601 aagcctttcg gccccggcaa gtttctcggt gactgatatg aaagaccaaa aggacaagca
     3661 gaccggcgac ctgctggcca gccctgacgc tgtacgccaa gcgcgatatg ccgagcgcat
     3721 gaaggccaaa gggatgcgtc agcgcaagtt ctggctgacc gacgacgaat acgaggcgct
     3781 gcgcgagtgc ctggaagaac tcagagcggc gcagggcggg ggtagtgacc ccgccagcgc
     3841 ctaaccacca actgcctgca aaggaggcaa tcaatggcta cccataagcc tatcaatatt
     3901 ctggaggcgt tcgcagcagc gccgccaccg ctggactacg ttttgcccaa catggtggcc
     3961 ggtacggtcg gggcgctggt gtcgcccggt ggtgccggta aatccatgct ggccctgcaa
     4021 ctggccgcac agattgcagg cgggccggat ctgctggagg tgggcgaact gcccaccggc
     4081 ccggtgatct acctgcccgc cgaagacccg cccaccgcca ttcatcaccg cctgcacgcc
     4141 cttggggcgc acctcagcgc cgaggaacgg caagccgtgg ctgacggcct gctgatccag
     4201 ccgctgatcg gcagcctgcc caacatcatg gccccggagt ggttcgacgg cctcaagcgc
     4261 gccgccgagg gccgccgcct gatggtgctg gacacgctgc gccggttcca catcgaggaa
     4321 gaaaacgcca gcggccccat ggcccaggtc atcggtcgca tggaggccat cgccgccgat
     4381 accgggtgct ctatcgtgtt cctgcaccat gccagcaagg gcgcggccat gatgggcgca
     4441 ggcgaccagc agcaggccag ccggggcagc tcggtactgg tcgataacat ccgctggcag
     4501 tcctacctgt cgagcatgac cagcgccgag gccgaggaat ggggtgtgga cgacgaccag
     4561 cgccggttct tcgtccgctt cggtgtgagc aaggccaact atggcgcacc gttcgctgat
     4621 cggtggttca ggcggcatga cggcggggtg ctcaagcccg ccgtgctgga gaggcagcgc
     4681 aagagcaagg gggtgccccg tggtgaagcc taagaacaag cacagcctca gccacgtccg
     4741 gcacgacccg gcgcactgtc tggcccccgg cctgttccgt gccctcaagc ggggcgagcg
     4801 caagcgcagc aagctggacg tgacgtatga ctacggcgac ggcaagcgga tcgagttcag
     4861 cggcccggag ccgctgggcg ctgatgatct gcgcatcctg caagggctgg tggccatggc
     4921 tgggcctaat ggcctagtgc ttggcccgga acccaagacc gaaggcggac ggcagctccg
     4981 gctgttcctg gaacccaagt gggaggccgt caccgctgat gccatggtgg tcaaaggtag
     5041 ctatcgggcg ctggcaaagg aaatcggggc agaggtcgat agtggtgggg cgctcaagca
     5101 catacaggac tgcatcgagc gcctttggaa ggtatccatc atcgcccaga atggccgcaa
     5161 gcggcagggg tttcggctgc tgtcggagta cgccagcgac gaggcggacg ggcgcctgta
     5221 cgtggccctg aaccccttga tcgcgcaggc cgtcatgggt ggcggccagc atgtgcgcat
     5281 cagcatggac gaggtgcggg cgctggacag cgaaaccgcc cgcctgctgc accagcggct
     5341 gtgtggctgg atcgaccccg gcaaaaccgg caaggcttcc atagatacct tgtgcggcta
     5401 tgtctggccg tcagaggcca gtggttcgac catgcgcaag cgccgccagc gggtgcgcga
     5461 ggcgttgccg gagctggtcg cgctgggctg gacggtaacc gagttcgcgg cgggcaagta
     5521 cgacatcacc cggcccaagg cggcaggctg acccccccca ctctattgta aacaagacat
     5581 ttttatcttt tatattcaat ggcttatttt cctgctaatt ggtaatacca tgaaaaatac
     5641 catgctcaga aaaggcttaa caatattttg aaaaattgcc tactgagcgc tgccgcacag
     5701 ctccatagcg tttaagggca ccaataactg ccttaaaaaa attacgcccc gccctgccac
     5761 tcatcgcagt acggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5821 ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg caactgttgg
     5881 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     5941 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     6001 gccagtgact tctgttgagt taagaacgag tatcgagatg gcacatagcc ttgctcaaat
     6061 tggaatcagg tttgtgccaa taccagtaga aacagacgaa gaatccatgg ttaagaccca
     6121 ctttcacatt taagttgttt ttctaatccg catatgatca attcaaggcc gaataagaag
     6181 gctggctctg caccttggtg atcaaataat tcgatagctt gtcgtaataa tggcggcata
     6241 ctatcagtag taggtgtttc cctttcttct ttagcgactt gatgctcttg atcttccaat
     6301 acgcaaccta aagtaaaatg ccccacagcg ctgagtgcat ataatgcatt ctctagtgaa
     6361 aaaccttgtt ggcataaaaa ggctaattga ttttcgagag tttcatactg tttttctgta
     6421 ggccgtgtac ctaaatgtac ttttgctcca tcgcgatgac ttagtaaagc acatctaaaa
     6481 cttttagcgt tattacgtaa aaaatcttgc cagctttccc cttctaaagg gcaaaagtga
     6541 gtatggtgcc tatctaacat ctcaatggct aaggcgtcga gcaaagcccg cttatttttt
     6601 acatgccaat acaatgtagg ctgctctaca cctagcttct gggcgagttt acgggttgtt
     6661 aaaccttcga ttccgacctc attaagcagc tctaatgcgc tgttaatcac tttactttta
     6721 tctaatctag acatcattaa ttcctaattt ttgttgacac tctatcattg atagagttat
     6781 tttaccactc cctatcagtg atagagagaa ctgggaagag cgcagattaa agaggagaaa
     6841 tactagatgg cttcctccga agacgttatc aaagagttca tgcgtttcaa agttcgtatg
     6901 gaaggttccg ttaacggtca cgagttcgaa atcgaaggtg aaggtgaagg tcgtccgtac
     6961 gaaggtaccc agaccgctaa actgaaagtt accaaaggtg gtccgctgcc gttcgcttgg
     7021 gacatcctgt ccccgcagtt ccagtacggt tccaaagctt acgttaaaca cccggctgac
     7081 atcccggact acctgaaact gtccttcccg gaaggtttca aatgggaacg tgttatgaac
     7141 ttcgaagacg gtggtgttgt taccgttacc caggactcct ccctgcaaga cggtgagttc
     7201 atctacaaag ttaaactgcg tggtaccaac ttcccgtccg acggtccggt tatgcagaaa
     7261 aaaaccatgg gttgggaagc ttccaccgaa cgtatgtacc cggaagacgg tgctctgaaa
     7321 ggtgaaatca aaatgcgtct gaaactgaaa gacggtggtc actacgacgc tgaagttaaa
     7381 accacctaca tggctaaaaa accggttcag ctgccgggtg cttacaaaac cgacatcaaa
     7441 ctggacatca cctcccacaa cgaagactac accatcgttg aacagtacga acgtgctgaa
     7501 ggtcgtcact ccaccggtgc ttaataacgc tgatagtgct agtgtagatc gcgctttcca
     7561 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7621 tttgcgtatt gggcgcatgc ataaaaactg ttgtaattca ttaagcattc tgccgacatg
     7681 gaatccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     7741 ttgcgtataa tatttgccca tggacgcaca ccgtggaaac ggatgaaggc acgaacccag
     7801 ttgacataag cctgttcggt tcgtaaactg taatgcaagt agcgtatgcg ctcacgcaac
     7861 tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct
     7921 tgttatgact gtttttttgt acagtctatg cctcgggcat ccaagcagca agcgcgttac
     7981 gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca gcaacgatgt
     8041 tacgcagcag ggcagtcgcc ctaaaacaaa gttaggtggc tcaagtatgg gcatcattcg
     8101 cacatgtagg ctcggccctg accaagtcaa atccatgcgg gctgctcttg atcttttcgg
     8161 tcgtgagttc ggagacgtag ccacctactc ccaacatcag ccggactccg attacctcgg
     8221 gaacttgctc cgtagtaaga cattcatcgc gcttgctgcc ttcgaccaag aagcggttgt
     8281 tggcgctctc gcggcttacg ttctgcccaa gtttgagcag ccgcgtagtg agatctatat
     8341 ctatgatctc gcagtctccg gagagcaccg gaggcagggc attgccaccg cgctcatcaa
     8401 tctcctcaag catgaggcca acgcgcttgg tgcttatgtg atctacgtgc aagcagatta
     8461 cggtgacgat cccgcagtgg ctctctatac aaagttgggc atacgggaag aagtgatgca
     8521 ctttgatatc gacccaagta ccgccaccta acaattcgtt caagccgaga tcggcttccc
     8581 ggccgcggag ttgttcggta aattgtcaca acgccgca
//
