LOCUS       sequence-265765-        8240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Conjugative vector derived from pSHDY. Constitutive expression of 
            the repressor tetR, as well as aTc-inducible expression of mVenus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8240)
  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 8240)
  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"; 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-PL03-mVenus_tetR
FEATURES             Location/Qualifiers
     source          1..8240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(6..797)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    1003..1024
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    1046..1064
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      1903..1974
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1990..2017
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    2024..2044
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(2245..3096)
                     /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
                     TAECHVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(3083..3922)
                     /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(4433..5404)
                     /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            6643..6730
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      7072..7466
                     /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)"
     promoter        7512..7546
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             7563..8186
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /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="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(7636..7655)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      8187..8214
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 agacattatt tgccgactac cttggtgatc tcgcctttca cgtagtggac aaattcttcc
       61 aactgatctg cgcgcgaggc caagcgatct tcttcttgtc caagataagc ctgtctagct
      121 tcaagtatga cgggctgata ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca
      181 tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact
      241 acatttcgct catcgccagc ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt
      301 agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga gttcctccgc cgctggacct
      361 accaaggcaa cgctatgttc tcttgctttt gtcagcaaga tagccagatc aatgtcgatc
      421 gtggctggct cgaagatacc tgcaagaatg tcattgcgct gccattctcc aaattgcagt
      481 tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct
      541 acagcgcgga gaatctcgct ctctccaggg gaagccgaag tttccaaaag gtcgttgatc
      601 aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg taaccagcaa atcaatatca
      661 ctgtgtggct tcaggccgcc atccactgcg gagccgtaca aatgtacggc cagcaacgtc
      721 ggttcgagat ggcgctcgat gacgccaact acctctgata gttgagtcga tacttcggcg
      781 atcaccgctt ccctcatgat gtttaacttt gttttagggc gactgccctg ctgcgtaaca
      841 tcgttgctgc tccataacat caaacatcga cccacggcgt aacgcgcttg ctgcttggat
      901 gcccgaggca tagactgtac cccaaaaaaa cagtcataac aagccatgaa aaccgccact
      961 gcgccgttac caccgctgcg ttcggtcaag gttctggacc aggaattcgc ggccgcttct
     1021 agaggtacgc aattggcatg cgcattccct atcagtgata gagattgaca tccctatcag
     1081 tgatagatat aatggccact tactagagta gtggaggtta ctagatggct agcaaaggag
     1141 aggaactgtt taccggtgtg gtacccattt tagtagaatt ggatggtgat gtgaacggcc
     1201 acaagttcag cgtttccggg gaaggcgaag gggatgcaac ctatggaaag ctaactttga
     1261 aactcatttg cactaccggt aaactccccg tcccctggcc tacgttagtc acgaccctcg
     1321 gttatggtct gcaatgtttt gcccgttatc ccgatcatat gaaacaacac gatttcttta
     1381 aatctgccat gccagaggga tatgttcaag aacggacaat tttctttaag gatgatggta
     1441 attataaaac ccgtgcagaa gtcaaatttg aaggtgatac actcgtgaat cgcatcgaac
     1501 taaaaggtat tgactttaag gaggatggaa acattctcgg ccataagtta gaatacaatt
     1561 acaacagcca taatgtgtac attactgcgg ataaacaaaa aaacggtatc aaagccaact
     1621 ttaaaattcg acataacatc gaagatggtg gcgttcagtt agcggatcac taccaacaaa
     1681 atactcccat tggagacgga ccagtcctgc tgcctgataa ccattatctg tcttatcaat
     1741 ccaaactgtc caaggatccg aatgaaaagc gggatcacat ggttctgttg gagtttgtga
     1801 ccgcggcagg tattactctc ggaatggacg aactatacaa aaggcctgcc gccaacgacg
     1861 aaaactacgc tgcatcagtt taatagtact agagccaggc atcaaataaa acgaaaggct
     1921 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag
     1981 agtcacactg gctcaccttc gggtgggcct ttctgcgttt atatactagt agcggccgct
     2041 gcaggagcag aagagcatac atctggaagc aaagccagga aagcggccta tggagctgtg
     2101 cggcagcgct cagtaggcaa tttttcaaaa tattgttaag ccttttctga gcatggtatt
     2161 tttcatggta ttaccaatta gcaggaaaat aagccattga atataaaaga taaaaatgtc
     2221 ttgtttacaa tagagtgggg ggggtcagcc tgccgccttg ggccgggtga tgtcgtactt
     2281 gcccgccgcg aactcggtta ccgtccagcc cagcgcgacc agctccggca acgcctcgcg
     2341 cacccgctgg cggcgcttgc gcatggtcga accactggcc tctgacggcc agacatagcc
     2401 gcacaaggta tctatggaag ccttgccggt tttgccgggg tcgatccagc cacacagccg
     2461 ctggtgcagc aggcgggcgg tttcgctgtc cagcgcccgc acctcgtcca tgctgatgcg
     2521 cacatgctgg ccgccaccca tgacggcctg cgcgatcaag gggttcaggg ccacgtacag
     2581 gcgcccgtcc gcctcgtcgc tggcgtactc cgacagcagc cgaaacccct gccgcttgcg
     2641 gccattctgg gcgatgatgg ataccttcca aaggcgctcg atgcagtcct gtatgtgctt
     2701 gagcgcccca ccactatcga cctctgcccc gatttccttt gccagcgccc gatagctacc
     2761 tttgaccaca tggcattcag cggtgacggc ctcccacttg ggttccagga acagccggag
     2821 ctgccgtccg ccttcggtct tgggttccgg gccaagcact aggccattag gcccagccat
     2881 ggccaccagc ccttgcagga tgcgcagatc atcagcgccc agcggctccg ggccgctgaa
     2941 ctcgatccgc ttgccgtcgc cgtagtcata cgtcacgtcc agcttgctgc gcttgcgctc
     3001 gccccgcttg agggcacgga acaggccggg ggccagacag tgcgccgggt cgtgccggac
     3061 gtggctgagg ctgtgcttgt tcttaggctt caccacgggg cacccccttg ctcttgcgct
     3121 gcctctccag cacggcgggc ttgagcaccc cgccgtcatg ccgcctgaac caccgatcag
     3181 cgaacggtgc gccatagttg gccttgctca caccgaagcg gacgaagaac cggcgctggt
     3241 cgtcgtccac accccattcc tcggcctcgg cgctggtcat gctcgacagg taggactgcc
     3301 agcggatgtt atcgaccagt accgagctgc cccggctggc ctgctgctgg tcgcctgcgc
     3361 ccatcatggc cgcgcccttg ctggcatggt gcaggaacac gatagagcac ccggtatcgg
     3421 cggcgatggc ctccatgcga ccgatgacct gggccatggg gccgctggcg ttttcttcct
     3481 cgatgtggaa ccggcgcagc gtgtccagca ccatcaggcg gcggccctcg gcggcgcgct
     3541 tgaggccgtc gaaccactcc ggggccatga tgttgggcag gctgccgatc agcggctgga
     3601 tcagcaggcc gtcagccacg gcttgccgtt cctcggcgct gaggtgcgcc ccaagggcgt
     3661 gcaggcggtg atgaatggcg gtgggcgggt cttcggcggg caggtagatc accgggccgg
     3721 tgggcagttc gcccacctcc agcagatccg gcccgcctgc aatctgtgcg gccagttgca
     3781 gggccagcat ggatttaccg gcaccaccgg gcgacaccag cgccccgacc gtaccggcca
     3841 ccatgttggg caaaacgtag tccagcggtg gcggcgctgc tgcgaacgcc tccagaatat
     3901 tgataggctt atgggtagcc attgattgcc tcctttgcag gcagttggtg gttaggcgct
     3961 ggcggggtca ctacccccgc cctgcgccgc tctgagttct tccaggcact cgcgcagcgc
     4021 ctcgtattcg tcgtcggtca gccagaactt gcgctgacgc atccctttgg ccttcatgcg
     4081 ctcggcatat cgcgcttggc gtacagcgtc agggctggcc agcaggtcgc cggtctgctt
     4141 gtccttttgg tctttcatat cagtcaccga gaaacttgcc ggggccgaaa ggcttgtctt
     4201 cgcggaacaa ggacaaggtg cagccgtcaa ggttaaggct ggccatatca gcgactgaaa
     4261 agcggccagc ctcggccttg tttgacgtat aaccaaagcc accgggcaac caatagccct
     4321 tgtcactttt gatcaggtag accgaccctg aagcgctttt ttcgtattcc ataaaacccc
     4381 cttctgtgcg tgagtactca tagtataaca ggcgtgagta ccaacgcaag cactacatgc
     4441 tgaaatctgg cccgcccctg tccatgcctc gctggcgggg tgccggtgcc cgtgccagct
     4501 cggcccgcgc aagctggacg ctgggcagac ccatgacctt gctgacggtg cgctcgatgt
     4561 aatccgcttc gtggccgggc ttgcgctctg ccagcgctgg gctggcctcg gccatggcct
     4621 tgccgatttc ctcggcactg cggccccggc tggccagctt ctgcgcggcg ataaagtcgc
     4681 acttgctgag gtcatcaccg aagcgcttga ccagcccggc catctcgctg cggtactcgt
     4741 ccagcgccgt gcgccggtgg cggctaagct gccgctcggg cagttcgagg ctggccagcc
     4801 tgcgggcctt ctcctgctgc cgctgggcct gctcgatctg ctggccagcc tgctgcacca
     4861 gcgccgggcc agcggtggcg gtcttgccct tggattcacg cagcagcacc cacggctgat
     4921 aaccggcgcg ggtggtgtgc ttgtccttgc ggttggtgaa gcccgccaag cggccatagt
     4981 ggcggctgtc ggcgctggcc gggtcggcgt cgtactcgct ggccagcgtc cgggcaatct
     5041 gcccccgaag ttcaccgcct gcggcgtcgg ccaccttgac ccatgcctga tagttcttcg
     5101 ggctggtttc cactaccagg gcaggctccc ggccctcggc tttcatgtca tccaggtcaa
     5161 actcgctgag gtcgtccacc agcaccagac catgccgctc ctgctcggcg ggcctgatat
     5221 acacgtcatt gccctgggca ttcatccgct tgagccatgg cgtgttctgg agcacttcgg
     5281 cggctgacca ttcccggttc atcatctggc cggtggtggc gtccctgacg ccgatatcga
     5341 agcgctcaca gcccatggcc ttgagctgtc ggcctatggc ctgcaaagtc ctgtcgttct
     5401 tcatcgggcc accaagcgca gccagatcga gccgtcctcg gttgtcagtg gcgtcaggtc
     5461 gagcaagagc aacgatgcga tcagcagcac caccgtaggc atcatggaag ccagcatcac
     5521 ggttagccat agcttccagt gccacccccg cgacgcgctc cgggcgctct gcgcggcgct
     5581 gctcacctcg gcggctacct cccgcaactc tttggccagc tccacccatg ccgcccctgt
     5641 ctggcgctgg gctttcagcc actccgccgc ctgcgcctcg ctggcctgct gggtctggct
     5701 catgacctgc cgggcttcgt cggccagtgt cgccatgctc tgggccagcg gttcgatctg
     5761 ctccgctaac tcgttgatgc ctctggattt cttcactctg tcgattgcgt tcatggtcta
     5821 ttgcctcccg gtattcctgt aagtcgatga tctgggcgtt ggcggtgtcg atgttcaggg
     5881 ccacgtctgc ccggtcggtg cggatgcccc ggccttccat ctccaccacg ttcggcccca
     5941 ggtgaacacc gggcaggcgc tcgatgccct gcgcctcaag tgttctgtgg tcaatgcggg
     6001 cgtcgtggcc agcccgctct aatgcccggt tggcatggtc ggcccatgcc tcgcgggtct
     6061 gctcaagcca tgccttgggc ttgagcgctt cggtcttctg tgccccgccc ttctccgggg
     6121 tcttgccgtt gtaccgcttg aaccactgag cggcgggccg ctcgatgccg tcattgatcc
     6181 gctcggagat catcaggtgg cagtgcgggt tctcgccgcc accggcatgg atggccagcg
     6241 tatacggcag gcgctcggca ccggtcaggt gctgggcgaa ctcggacgcc agcgccttct
     6301 gctggtcgag ggtcagctcg accggcaggg caaattcgac ctccttgaac agccgcccat
     6361 tggcgcgttc atacaggtcg gcagcatccc agtagtcggc gggccgctcg acgaactccg
     6421 gcatgtgccc ggattcggcg tgcaagactt catccatgtc gcgggcatac ttgccttcgc
     6481 gctggatgaa gtcggccttg gccctggccg attggccgcc cgacctgctg ccggttttcg
     6541 ccgtaaggtg ataaatcgcc atgctgcctc gctgttgctt ttgcttttcg gctccatgca
     6601 atggccctcg gagagcgcac cgcccgaagg gtggccgtta ggccagtttc tcgaagagaa
     6661 accggtaagt gcgccctccc ctacaaagta gggtcgggat tgccgccgct gtgcctccat
     6721 gatagcctac gagacagcac attaacaatg gggtgtcaag atggttaagg ggagcaacaa
     6781 ggcggcggat cggctggcca agctcgaaga acaacgagcg cgaatcaatg ccgaaattca
     6841 gcgggtgcgg gcaagggaac agcagcaaga gcgcaagaac gaaacaaggc gcaaggtgct
     6901 ggtgggggcc atgattttgg ccaaggtgaa cagcagcgag tggccggagg atcggctcat
     6961 ggcggcaatg gatgcgtacc ttgaacgcga ccacgaccgc gccttgttcg gtctgccgcc
     7021 acgccagaag gatgagccgg gctgaatgat cgaccgagac aggccctgcg gggctgcaca
     7081 cgcgccccca cccttcgggt agggggaaag gccgctaaag cggctaaaag cgctccagcg
     7141 tatttctgcg gggtttggtg tggggtttag cgggctttgc ccgcctttcc ccctgccgcg
     7201 cagcggtggg gcggtgtgta gcctagcgca gcgaatagac cagctatccg gcctctggcc
     7261 gggcatattg ggcaagggca gcagcgcccc acaagggcgc tgataaccgc gcctagtgga
     7321 ttattcttag ataatcatgg atggattttt ccaacacccc gccagccccc gcccctgctg
     7381 ggtttgcagg tttgggggcg tgacagttat tgcaggggtt cgtgacagtt attgcagggg
     7441 ggcgtgacag ttattgcagg ggttcgtgac agttagtacg ggagtgacgg gggtaccgag
     7501 tcgatgtcga cttgacagct agctcagtcc taggtataat gctagcaaga gagaaatact
     7561 agatgtccag attagataaa agtaaagtga ttaacagcgc attagagctg cttaatgagg
     7621 tcggaatcga aggtttaaca acccgtaaac tcgcccagaa gctaggtgta gagcagccta
     7681 cattgtattg gcatgtaaaa aataagcggg ctttgctcga cgccttagcc attgagatgt
     7741 tagataggca ccatactcac ttttgccctt tagaagggga aagctggcaa gattttttac
     7801 gtaataacgc taaaagtttt agatgtgctt tactaagtca tcgcgatgga gcaaaagtac
     7861 atttaggtac acggcctaca gaaaaacagt atgaaactct cgaaaatcaa ttagcctttt
     7921 tatgccaaca aggtttttca ctagagaatg cattatatgc actcagcgct gtggggcatt
     7981 ttactttagg ttgcgtattg gaagatcaag agcatcaagt cgctaaagaa gaaagggaaa
     8041 cacctactac tgatagtatg ccgccattat tacgacaagc tatcgaatta tttgatcacc
     8101 aaggtgcaga gccagccttc ttattcggcc ttgaattgat catatgcgga ttagaaaaac
     8161 aacttaaatg tgaaagtggg tcctaaggct caccttcggg tgggcctttc tgcgtttata
     8221 ctcgagagcg gccgaagctt
//
