LOCUS       sequence-273042-        8632 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Conjugative vector derived from pSHDY. Constitutive expression of 
            the activator rhaS, as well as rhamnose-inducible expression of 
            mVenus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8632)
  AUTHORS   Behle A, Saake P, Germann AT, Dienst D, Axmann IM
  TITLE     Comparative Dose-Response Analysis of Inducible Promoters in 
            Cyanobacteria.
  JOURNAL   ACS Synth Biol. 2020 Apr 17;9(4):843-855. doi: 
            10.1021/acssynbio.9b00505. Epub 2020 Mar 17.
  PUBMED    32134640
REFERENCE   2  (bases 1 to 8632)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.9b00505"; journalName: "ACS Synth Biol"; date: 
            "2020-04-17- 17"; volume: "9"; issue: "4"; pages: "843-855"
COMMENT     Sequence Label: pSHDY-Prha-mVenus_rhaS
FEATURES             Location/Qualifiers
     source          1..8632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        365..399
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     RBS             400..411
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             418..1254
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter
                     "
                     /label=rhaS
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQLYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     terminator      1299..1370
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1386..1413
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1445..2236)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVMHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    2442..2463
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      3388..3459
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3475..3502
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3509..3529
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(3730..4581)
                     /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(4568..5407)
                     /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(5918..6889)
                     /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"
     oriT            8128..8215
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      join(8557..8632,1..319)
                     /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)"
ORIGIN
        1 gcggggtttg gtgtggggtt tagcgggctt tgcccgcctt tccccctgcc gcgcagcggt
       61 ggggcggtgt gtagcctagc gcagcgaata gaccagctat ccggcctctg gccgggcata
      121 ttgggcaagg gcagcagcgc cccacaaggg cgctgataac cgcgcctagt ggattattct
      181 tagataatca tggatggatt tttccaacac cccgccagcc cccgcccctg ctgggtttgc
      241 aggtttgggg gcgtgacagt tattgcaggg gttcgtgaca gttattgcag gggggcgtga
      301 cagttattgc aggggttcgt gacagttagt acgggagtga cgggggtacc gagtcgatgt
      361 cgacttgaca gctagctcag tcctaggtat aatgctagca aagaggagaa atactagatg
      421 accgtattac atagtgtgga tttttttccg tctggtaacg cgtccgtggc gatagaaccc
      481 cggctcccgc aggcggattt tcctgaacat catcatgatt ttcatgaaat tgtgattgtc
      541 gaacatggca cgggtattca tgtgtttaat gggcagccct ataccatcac cggtggcacg
      601 gtctgtttcg tacgcgatca tgatcggcat ctgtatgaac ataccgataa tctgtgtctg
      661 accaatgtgc tgtatcgctc gccggatcga tttcagtttc tcgccgggct gaatcagttg
      721 ctgccacaag agctggatgg gcagtatccg tctcactggc gcgttaacca cagcgtattg
      781 cagcaggtgc gacagctggt tgcacagatg gaacagcagg aaggggaaaa tgatttaccc
      841 tcgaccgcca gtcgcgagat cttgtttatg caattactgc tcttgctgcg taaaagcagt
      901 ttgcaggaga acctggaaaa cagcgcatca cgtctcaact tgcttctggc ctggctggag
      961 gaccattttg ccgatgaggt gaattgggat gccgtggcgg atcaattttc tctttcactg
     1021 cgtacgctac atcggcagct taagcagcaa acgggactga cgcctcagct atacctgaac
     1081 cgcctgcgac tgatgaaagc ccgacatctg ctacgccaca gcgaggccag cgttactgac
     1141 atcgcctatc gctgtggatt cagcgacagt aaccactttt cgacgctttt tcgccgagag
     1201 tttaactggt caccgcgtga tattcgccag ggacgggatg gctttctgca ataataacac
     1261 tgatagtgct agtgtagatc actactagag ccaggcatca aataaaacga aaggctcagt
     1321 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc
     1381 acactggctc accttcgggt gggcctttct gcgtttatac tcgagagcgg ccgaagctta
     1441 gacattattt gccgactacc ttggtgatct cgcctttcac gtagtggaca aattcttcca
     1501 actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc tgtctagctt
     1561 caagtatgac gggctgatac tgggccggca ggcgctccat tgcccagtcg gcagcgacat
     1621 ccttcggcgc gattttgccg gttactgcgc tgtaccaaat gcgggacaac gtaagcacta
     1681 catttcgctc atcgccagcc cagtcgggcg gcgagttcca tagcgttaag gtttcattta
     1741 gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc gctggaccta
     1801 ccaaggcaac gctatgttct cttgcttttg tcagcaagat agccagatca atgtcgatcg
     1861 tggctggctc gaagatacct gcaagaatgt cattgcgctg ccattctcca aattgcagtt
     1921 cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg cataacaatg gtgacttcta
     1981 cagcgcggag aatctcgctc tctccagggg aagccgaagt ttccaaaagg tcgttgatca
     2041 aagctcgccg cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac
     2101 tgtgtggctt caggccgcca tccactgcgg agccgtacaa atgtacggcc agcaacgtcg
     2161 gttcgagatg gcgctcgatg acgccaacta cctctgatag ttgagtcgat acttcggcga
     2221 tcaccgcttc cctcatgatg tttaactttg ttttagggcg actgccctgc tgcgtaacat
     2281 cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg
     2341 cccgaggcat agactgtacc ccaaaaaaac agtcataaca agccatgaaa accgccactg
     2401 cgccgttacc accgctgcgt tcggtcaagg ttctggacca ggaattcgcg gccgcttcta
     2461 gagccacaat tcagcaaatt gtgaacatca tcacgttcat ctttccctgg ttgccaatgg
     2521 cccattttcc tgtcagtaac gagaaggtcg cgaattcagg cgctttttag actggtcgta
     2581 atgaatacta gagtagtgga ggttactaga tggctagcaa aggagaggaa ctgtttaccg
     2641 gtgtggtacc cattttagta gaattggatg gtgatgtgaa cggccacaag ttcagcgttt
     2701 ccggggaagg cgaaggggat gcaacctatg gaaagctaac tttgaaactc atttgcacta
     2761 ccggtaaact ccccgtcccc tggcctacgt tagtcacgac cctcggttat ggtctgcaat
     2821 gttttgcccg ttatcccgat catatgaaac aacacgattt ctttaaatct gccatgccag
     2881 agggatatgt tcaagaacgg acaattttct ttaaggatga tggtaattat aaaacccgtg
     2941 cagaagtcaa atttgaaggt gatacactcg tgaatcgcat cgaactaaaa ggtattgact
     3001 ttaaggagga tggaaacatt ctcggccata agttagaata caattacaac agccataatg
     3061 tgtacattac tgcggataaa caaaaaaacg gtatcaaagc caactttaaa attcgacata
     3121 acatcgaaga tggtggcgtt cagttagcgg atcactacca acaaaatact cccattggag
     3181 acggaccagt cctgctgcct gataaccatt atctgtctta tcaatccaaa ctgtccaagg
     3241 atccgaatga aaagcgggat cacatggttc tgttggagtt tgtgaccgcg gcaggtatta
     3301 ctctcggaat ggacgaacta tacaaaaggc ctgccgccaa cgacgaaaac tacgctgcat
     3361 cagtttaata gtactagagc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     3421 gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca
     3481 ccttcgggtg ggcctttctg cgtttatata ctagtagcgg ccgctgcagg agcagaagag
     3541 catacatctg gaagcaaagc caggaaagcg gcctatggag ctgtgcggca gcgctcagta
     3601 ggcaattttt caaaatattg ttaagccttt tctgagcatg gtatttttca tggtattacc
     3661 aattagcagg aaaataagcc attgaatata aaagataaaa atgtcttgtt tacaatagag
     3721 tggggggggt cagcctgccg ccttgggccg ggtgatgtcg tacttgcccg ccgcgaactc
     3781 ggttaccgtc cagcccagcg cgaccagctc cggcaacgcc tcgcgcaccc gctggcggcg
     3841 cttgcgcatg gtcgaaccac tggcctctga cggccagaca tagccgcaca aggtatctat
     3901 ggaagccttg ccggttttgc cggggtcgat ccagccacac agccgctggt gcagcaggcg
     3961 ggcggtttcg ctgtccagcg cccgcacctc gtccatgctg atgcgcacat gctggccgcc
     4021 acccatgacg gcctgcgcga tcaaggggtt cagggccacg tacaggcgcc cgtccgcctc
     4081 gtcgctggcg tactccgaca gcagccgaaa cccctgccgc ttgcggccat tctgggcgat
     4141 gatggatacc ttccaaaggc gctcgatgca gtcctgtatg tgcttgagcg ccccaccact
     4201 atcgacctct gccccgattt cctttgccag cgcccgatag ctacctttga ccaccatggc
     4261 atcagcggtg acggcctccc acttgggttc caggaacagc cggagctgcc gtccgccttc
     4321 ggtcttgggt tccgggccaa gcactaggcc attaggccca gccatggcca ccagcccttg
     4381 caggatgcgc agatcatcag cgcccagcgg ctccgggccg ctgaactcga tccgcttgcc
     4441 gtcgccgtag tcatacgtca cgtccagctt gctgcgcttg cgctcgcccc gcttgagggc
     4501 acggaacagg ccgggggcca gacagtgcgc cgggtcgtgc cggacgtggc tgaggctgtg
     4561 cttgttctta ggcttcacca cggggcaccc ccttgctctt gcgctgcctc tccagcacgg
     4621 cgggcttgag caccccgccg tcatgccgcc tgaaccaccg atcagcgaac ggtgcgccat
     4681 agttggcctt gctcacaccg aagcggacga agaaccggcg ctggtcgtcg tccacacccc
     4741 attcctcggc ctcggcgctg gtcatgctcg acaggtagga ctgccagcgg atgttatcga
     4801 ccagtaccga gctgccccgg ctggcctgct gctggtcgcc tgcgcccatc atggccgcgc
     4861 ccttgctggc atggtgcagg aacacgatag agcacccggt atcggcggcg atggcctcca
     4921 tgcgaccgat gacctgggcc atggggccgc tggcgttttc ttcctcgatg tggaaccggc
     4981 gcagcgtgtc cagcaccatc aggcggcggc cctcggcggc gcgcttgagg ccgtcgaacc
     5041 actccggggc catgatgttg ggcaggctgc cgatcagcgg ctggatcagc aggccgtcag
     5101 ccacggcttg ccgttcctcg gcgctgaggt gcgccccaag ggcgtgcagg cggtgatgaa
     5161 tggcggtggg cgggtcttcg gcgggcaggt agatcaccgg gccggtgggc agttcgccca
     5221 cctccagcag atccggcccg cctgcaatct gtgcggccag ttgcagggcc agcatggatt
     5281 taccggcacc accgggcgac accagcgccc cgaccgtacc ggccaccatg ttgggcaaaa
     5341 cgtagtccag cggtggcggc gctgctgcga acgcctccag aatattgata ggcttatggg
     5401 tagccattga ttgcctcctt tgcaggcagt tggtggttag gcgctggcgg ggtcactacc
     5461 cccgccctgc gccgctctga gttcttccag gcactcgcgc agcgcctcgt attcgtcgtc
     5521 ggtcagccag aacttgcgct gacgcatccc tttggccttc atgcgctcgg catatcgcgc
     5581 ttggcgtaca gcgtcagggc tggccagcag gtcgccggtc tgcttgtcct tttggtcttt
     5641 catatcagtc accgagaaac ttgccggggc cgaaaggctt gtcttcgcgg aacaaggaca
     5701 aggtgcagcc gtcaaggtta aggctggcca tatcagcgac tgaaaagcgg ccagcctcgg
     5761 ccttgtttga cgtataacca aagccaccgg gcaaccaata gcccttgtca cttttgatca
     5821 ggtagaccga ccctgaagcg cttttttcgt attccataaa acccccttct gtgcgtgagt
     5881 actcatagta taacaggcgt gagtaccaac gcaagcacta catgctgaaa tctggcccgc
     5941 ccctgtccat gcctcgctgg cggggtgccg gtgcccgtgc cagctcggcc cgcgcaagct
     6001 ggacgctggg cagacccatg accttgctga cggtgcgctc gatgtaatcc gcttcgtggc
     6061 cgggcttgcg ctctgccagc gctgggctgg cctcggccat ggccttgccg atttcctcgg
     6121 cactgcggcc ccggctggcc agcttctgcg cggcgataaa gtcgcacttg ctgaggtcat
     6181 caccgaagcg cttgaccagc ccggccatct cgctgcggta ctcgtccagc gccgtgcgcc
     6241 ggtggcggct aagctgccgc tcgggcagtt cgaggctggc cagcctgcgg gccttctcct
     6301 gctgccgctg ggcctgctcg atctgctggc cagcctgctg caccagcgcc gggccagcgg
     6361 tggcggtctt gcccttggat tcacgcagca gcacccacgg ctgataaccg gcgcgggtgg
     6421 tgtgcttgtc cttgcggttg gtgaagcccg ccaagcggcc atagtggcgg ctgtcggcgc
     6481 tggccgggtc ggcgtcgtac tcgctggcca gcgtccgggc aatctgcccc cgaagttcac
     6541 cgcctgcggc gtcggccacc ttgacccatg cctgatagtt cttcgggctg gtttccacta
     6601 ccagggcagg ctcccggccc tcggctttca tgtcatccag gtcaaactcg ctgaggtcgt
     6661 ccaccagcac cagaccatgc cgctcctgct cggcgggcct gatatacacg tcattgccct
     6721 gggcattcat ccgcttgagc catggcgtgt tctggagcac ttcggcggct gaccattccc
     6781 ggttcatcat ctggccggtg gtggcgtccc tgacgccgat atcgaagcgc tcacagccca
     6841 tggccttgag ctgtcggcct atggcctgca aagtcctgtc gttcttcatc gggccaccaa
     6901 gcgcagccag atcgagccgt cctcggttgt cagtggcgtc aggtcgagca agagcaacga
     6961 tgcgatcagc agcaccaccg taggcatcat ggaagccagc atcacggtta gccatagctt
     7021 ccagtgccac ccccgcgacg cgctccgggc gctctgcgcg gcgctgctca cctcggcggc
     7081 tacctcccgc aactctttgg ccagctccac ccatgccgcc cctgtctggc gctgggcttt
     7141 cagccactcc gccgcctgcg cctcgctggc ctgctgggtc tggctcatga cctgccgggc
     7201 ttcgtcggcc agtgtcgcca tgctctgggc cagcggttcg atctgctccg ctaactcgtt
     7261 gatgcctctg gatttcttca ctctgtcgat tgcgttcatg gtctattgcc tcccggtatt
     7321 cctgtaagtc gatgatctgg gcgttggcgg tgtcgatgtt cagggccacg tctgcccggt
     7381 cggtgcggat gccccggcct tccatctcca ccacgttcgg ccccaggtga acaccgggca
     7441 ggcgctcgat gccctgcgcc tcaagtgttc tgtggtcaat gcgggcgtcg tggccagccc
     7501 gctctaatgc ccggttggca tggtcggccc atgcctcgcg ggtctgctca agccatgcct
     7561 tgggcttgag cgcttcggtc ttctgtgccc cgcccttctc cggggtcttg ccgttgtacc
     7621 gcttgaacca ctgagcggcg ggccgctcga tgccgtcatt gatccgctcg gagatcatca
     7681 ggtggcagtg cgggttctcg ccgccaccgg catggatggc cagcgtatac ggcaggcgct
     7741 cggcaccggt caggtgctgg gcgaactcgg acgccagcgc cttctgctgg tcgagggtca
     7801 gctcgaccgg cagggcaaat tcgacctcct tgaacagccg cccattggcg cgttcataca
     7861 ggtcggcagc atcccagtag tcggcgggcc gctcgacgaa ctccggcatg tgcccggatt
     7921 cggcgtgcaa gacttcatcc atgtcgcggg catacttgcc ttcgcgctgg atgaagtcgg
     7981 ccttggccct ggccgattgg ccgcccgacc tgctgccggt tttcgccgta aggtgataaa
     8041 tcgccatgct gcctcgctgt tgcttttgct tttcggctcc atgcaatggc cctcggagag
     8101 cgcaccgccc gaagggtggc cgttaggcca gtttctcgaa gagaaaccgg taagtgcgcc
     8161 ctcccctaca aagtagggtc gggattgccg ccgctgtgcc tccatgatag cctacgagac
     8221 agcacattaa caatggggtg tcaagatggt taaggggagc aacaaggcgg cggatcggct
     8281 ggccaagctc gaagaacaac gagcgcgaat caatgccgaa attcagcggg tgcgggcaag
     8341 ggaacagcag caagagcgca agaacgaaac aaggcgcaag gtgctggtgg gggccatgat
     8401 tttggccaag gtgaacagca gcgagtggcc ggaggatcgg ctcatggcgg caatggatgc
     8461 gtaccttgaa cgcgaccacg accgcgcctt gttcggtctg ccgccacgcc agaaggatga
     8521 gccgggctga atgatcgacc gagacaggcc ctgcggggct gcacacgcgc ccccaccctt
     8581 cgggtagggg gaaaggccgc taaagcggct aaaagcgctc cagcgtattt ct
//
