LOCUS       sequence-273652-        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"
     misc_feature    70..91
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    113..131
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      970..1041
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1057..1084
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1091..1111
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(1312..2163)
                     /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(2150..2989)
                     /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(3500..4471)
                     /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            5710..5797
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      6139..6533
                     /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        6579..6613
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             6630..7253
                     /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(6703..6722)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      7254..7281
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(7313..8104)
                     /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"
ORIGIN
        1 tcataacaag ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt
       61 ctggaccagg aattcgcggc cgcttctaga ggtacgcaat tggcatgcgc attccctatc
      121 agtgatagag attgacatcc ctatcagtga tagatataat ggccacttac tagagtagtg
      181 gaggttacta gatggctagc aaaggagagg aactgtttac cggtgtggta cccattttag
      241 tagaattgga tggtgatgtg aacggccaca agttcagcgt ttccggggaa ggcgaagggg
      301 atgcaaccta tggaaagcta actttgaaac tcatttgcac taccggtaaa ctccccgtcc
      361 cctggcctac gttagtcacg accctcggtt atggtctgca atgttttgcc cgttatcccg
      421 atcatatgaa acaacacgat ttctttaaat ctgccatgcc agagggatat gttcaagaac
      481 ggacaatttt ctttaaggat gatggtaatt ataaaacccg tgcagaagtc aaatttgaag
      541 gtgatacact cgtgaatcgc atcgaactaa aaggtattga ctttaaggag gatggaaaca
      601 ttctcggcca taagttagaa tacaattaca acagccataa tgtgtacatt actgcggata
      661 aacaaaaaaa cggtatcaaa gccaacttta aaattcgaca taacatcgaa gatggtggcg
      721 ttcagttagc ggatcactac caacaaaata ctcccattgg agacggacca gtcctgctgc
      781 ctgataacca ttatctgtct tatcaatcca aactgtccaa ggatccgaat gaaaagcggg
      841 atcacatggt tctgttggag tttgtgaccg cggcaggtat tactctcgga atggacgaac
      901 tatacaaaag gcctgccgcc aacgacgaaa actacgctgc atcagtttaa tagtactaga
      961 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     1021 tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc
     1081 tgcgtttata tactagtagc ggccgctgca ggagcagaag agcatacatc tggaagcaaa
     1141 gccaggaaag cggcctatgg agctgtgcgg cagcgctcag taggcaattt ttcaaaatat
     1201 tgttaagcct tttctgagca tggtattttt catggtatta ccaattagca ggaaaataag
     1261 ccattgaata taaaagataa aaatgtcttg tttacaatag agtggggggg gtcagcctgc
     1321 cgccttgggc cgggtgatgt cgtacttgcc cgccgcgaac tcggttaccg tccagcccag
     1381 cgcgaccagc tccggcaacg cctcgcgcac ccgctggcgg cgcttgcgca tggtcgaacc
     1441 actggcctct gacggccaga catagccgca caaggtatct atggaagcct tgccggtttt
     1501 gccggggtcg atccagccac acagccgctg gtgcagcagg cgggcggttt cgctgtccag
     1561 cgcccgcacc tcgtccatgc tgatgcgcac atgctggccg ccacccatga cggcctgcgc
     1621 gatcaagggg ttcagggcca cgtacaggcg cccgtccgcc tcgtcgctgg cgtactccga
     1681 cagcagccga aacccctgcc gcttgcggcc attctgggcg atgatggata ccttccaaag
     1741 gcgctcgatg cagtcctgta tgtgcttgag cgccccacca ctatcgacct ctgccccgat
     1801 ttcctttgcc agcgcccgat agctaccttt gaccaccatg gcatcagcgg tgacggcctc
     1861 ccacttgggt tccaggaaca gccggagctg ccgtccgcct tcggtcttgg gttccgggcc
     1921 aagcactagg ccattaggcc cagccatggc caccagccct tgcaggatgc gcagatcatc
     1981 agcgcccagc ggctccgggc cgctgaactc gatccgcttg ccgtcgccgt agtcatacgt
     2041 cacgtccagc ttgctgcgct tgcgctcgcc ccgcttgagg gcacggaaca ggccgggggc
     2101 cagacagtgc gccgggtcgt gccggacgtg gctgaggctg tgcttgttct taggcttcac
     2161 cacggggcac ccccttgctc ttgcgctgcc tctccagcac ggcgggcttg agcaccccgc
     2221 cgtcatgccg cctgaaccac cgatcagcga acggtgcgcc atagttggcc ttgctcacac
     2281 cgaagcggac gaagaaccgg cgctggtcgt cgtccacacc ccattcctcg gcctcggcgc
     2341 tggtcatgct cgacaggtag gactgccagc ggatgttatc gaccagtacc gagctgcccc
     2401 ggctggcctg ctgctggtcg cctgcgccca tcatggccgc gcccttgctg gcatggtgca
     2461 ggaacacgat agagcacccg gtatcggcgg cgatggcctc catgcgaccg atgacctggg
     2521 ccatggggcc gctggcgttt tcttcctcga tgtggaaccg gcgcagcgtg tccagcacca
     2581 tcaggcggcg gccctcggcg gcgcgcttga ggccgtcgaa ccactccggg gccatgatgt
     2641 tgggcaggct gccgatcagc ggctggatca gcaggccgtc agccacggct tgccgttcct
     2701 cggcgctgag gtgcgcccca agggcgtgca ggcggtgatg aatggcggtg ggcgggtctt
     2761 cggcgggcag gtagatcacc gggccggtgg gcagttcgcc cacctccagc agatccggcc
     2821 cgcctgcaat ctgtgcggcc agttgcaggg ccagcatgga tttaccggca ccaccgggcg
     2881 acaccagcgc cccgaccgta ccggccacca tgttgggcaa aacgtagtcc agcggtggcg
     2941 gcgctgctgc gaacgcctcc agaatattga taggcttatg ggtagccatt gattgcctcc
     3001 tttgcaggca gttggtggtt aggcgctggc ggggtcacta cccccgccct gcgccgctct
     3061 gagttcttcc aggcactcgc gcagcgcctc gtattcgtcg tcggtcagcc agaacttgcg
     3121 ctgacgcatc cctttggcct tcatgcgctc ggcatatcgc gcttggcgta cagcgtcagg
     3181 gctggccagc aggtcgccgg tctgcttgtc cttttggtct ttcatatcag tcaccgagaa
     3241 acttgccggg gccgaaaggc ttgtcttcgc ggaacaagga caaggtgcag ccgtcaaggt
     3301 taaggctggc catatcagcg actgaaaagc ggccagcctc ggccttgttt gacgtataac
     3361 caaagccacc gggcaaccaa tagcccttgt cacttttgat caggtagacc gaccctgaag
     3421 cgcttttttc gtattccata aaaccccctt ctgtgcgtga gtactcatag tataacaggc
     3481 gtgagtacca acgcaagcac tacatgctga aatctggccc gcccctgtcc atgcctcgct
     3541 ggcggggtgc cggtgcccgt gccagctcgg cccgcgcaag ctggacgctg ggcagaccca
     3601 tgaccttgct gacggtgcgc tcgatgtaat ccgcttcgtg gccgggcttg cgctctgcca
     3661 gcgctgggct ggcctcggcc atggccttgc cgatttcctc ggcactgcgg ccccggctgg
     3721 ccagcttctg cgcggcgata aagtcgcact tgctgaggtc atcaccgaag cgcttgacca
     3781 gcccggccat ctcgctgcgg tactcgtcca gcgccgtgcg ccggtggcgg ctaagctgcc
     3841 gctcgggcag ttcgaggctg gccagcctgc gggccttctc ctgctgccgc tgggcctgct
     3901 cgatctgctg gccagcctgc tgcaccagcg ccgggccagc ggtggcggtc ttgcccttgg
     3961 attcacgcag cagcacccac ggctgataac cggcgcgggt ggtgtgcttg tccttgcggt
     4021 tggtgaagcc cgccaagcgg ccatagtggc ggctgtcggc gctggccggg tcggcgtcgt
     4081 actcgctggc cagcgtccgg gcaatctgcc cccgaagttc accgcctgcg gcgtcggcca
     4141 ccttgaccca tgcctgatag ttcttcgggc tggtttccac taccagggca ggctcccggc
     4201 cctcggcttt catgtcatcc aggtcaaact cgctgaggtc gtccaccagc accagaccat
     4261 gccgctcctg ctcggcgggc ctgatataca cgtcattgcc ctgggcattc atccgcttga
     4321 gccatggcgt gttctggagc acttcggcgg ctgaccattc ccggttcatc atctggccgg
     4381 tggtggcgtc cctgacgccg atatcgaagc gctcacagcc catggccttg agctgtcggc
     4441 ctatggcctg caaagtcctg tcgttcttca tcgggccacc aagcgcagcc agatcgagcc
     4501 gtcctcggtt gtcagtggcg tcaggtcgag caagagcaac gatgcgatca gcagcaccac
     4561 cgtaggcatc atggaagcca gcatcacggt tagccatagc ttccagtgcc acccccgcga
     4621 cgcgctccgg gcgctctgcg cggcgctgct cacctcggcg gctacctccc gcaactcttt
     4681 ggccagctcc acccatgccg cccctgtctg gcgctgggct ttcagccact ccgccgcctg
     4741 cgcctcgctg gcctgctggg tctggctcat gacctgccgg gcttcgtcgg ccagtgtcgc
     4801 catgctctgg gccagcggtt cgatctgctc cgctaactcg ttgatgcctc tggatttctt
     4861 cactctgtcg attgcgttca tggtctattg cctcccggta ttcctgtaag tcgatgatct
     4921 gggcgttggc ggtgtcgatg ttcagggcca cgtctgcccg gtcggtgcgg atgccccggc
     4981 cttccatctc caccacgttc ggccccaggt gaacaccggg caggcgctcg atgccctgcg
     5041 cctcaagtgt tctgtggtca atgcgggcgt cgtggccagc ccgctctaat gcccggttgg
     5101 catggtcggc ccatgcctcg cgggtctgct caagccatgc cttgggcttg agcgcttcgg
     5161 tcttctgtgc cccgcccttc tccggggtct tgccgttgta ccgcttgaac cactgagcgg
     5221 cgggccgctc gatgccgtca ttgatccgct cggagatcat caggtggcag tgcgggttct
     5281 cgccgccacc ggcatggatg gccagcgtat acggcaggcg ctcggcaccg gtcaggtgct
     5341 gggcgaactc ggacgccagc gccttctgct ggtcgagggt cagctcgacc ggcagggcaa
     5401 attcgacctc cttgaacagc cgcccattgg cgcgttcata caggtcggca gcatcccagt
     5461 agtcggcggg ccgctcgacg aactccggca tgtgcccgga ttcggcgtgc aagacttcat
     5521 ccatgtcgcg ggcatacttg ccttcgcgct ggatgtagtc ggccttggcc ctggccgatt
     5581 ggccgcccga cctgctgccg gttttcgccg taaggtgata aatcgccatg ctgcctcgct
     5641 gttgcttttg cttttcggct ccatgcaatg gccctcggag agcgcaccgc ccgaagggtg
     5701 gccgttaggc cagtttctcg aagagaaacc ggtaagtgcg ccctccccta caaagtaggg
     5761 tcgggattgc cgccgctgtg cctccatgat agcctacgag acagcacatt aacaatgggg
     5821 tgtcaagatg gttaagggga gcaacaaggc ggcggatcgg ctggccaagc tcgaagaaca
     5881 acgagcgcga atcaatgccg aaattcagcg ggtgcgggca agggaacagc agcaagagcg
     5941 caagaacgaa acaaggcgca aggtgctggt gggggccatg attttggcca aggtgaacag
     6001 cagcgagtgg ccggaggatc ggctcatggc ggcaatggat gcgtaccttg aacgcgacca
     6061 cgaccgcgcc ttgttcggtc tgccgccacg ccagaaggat gagccgggct gaatgatcga
     6121 ccgagacagg ccctgcgggg ctgcacacgc gcccccaccc ttcgggtagg gggaaaggcc
     6181 gctaaagcgg ctaaaagcgc tccagcgtat ttctgcgggg tttggtgtgg ggtttagcgg
     6241 gctttgcccg cctttccccc tgccgcgcag cggtggggcg gtgtgtagcc tagcgcagcg
     6301 aatagaccag ctatccggcc tctggccggg catattgggc aagggcagca gcgccccaca
     6361 agggcgctga taaccgcgcc tagtggatta ttcttagata atcatggatg gatttttcca
     6421 acaccccgcc agcccccgcc cctgctgggt ttgcaggttt gggggcgtga cagttattgc
     6481 aggggttcgt gacagttatt gcaggggggc gtgacagtta ttgcaggggt tcgtgacagt
     6541 tagtacggga gtgacggggg taccgagtcg atgtcgactt gacagctagc tcagtcctag
     6601 gtataatgct agcaagagag aaatactaga tgtccagatt agataaaagt aaagtgatta
     6661 acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg
     6721 cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat aagcgggctt
     6781 tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt tgccctttag
     6841 aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga tgtgctttac
     6901 taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa aaacagtatg
     6961 aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta gagaatgcat
     7021 tatatgcact cagcgctgtg gggcatttta ctttaggttg cgtattggaa gatcaagagc
     7081 atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg ccattattac
     7141 gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta ttcggccttg
     7201 aattgatcat atgcggatta gaaaaacaac ttaaatgtga aagtgggtcc taaggctcac
     7261 cttcgggtgg gcctttctgc gtttatactc gagagcggcc gaagcttaga cattatttgc
     7321 cgactacctt ggtgatctcg cctttcacgt agtggacaaa ttcttccaac tgatctgcgc
     7381 gcgaggccaa gcgatcttct tcttgtccaa gataagcctg tctagcttca agtatgacgg
     7441 gctgatactg ggccggcagg cgctccattg cccagtcggc agcgacatcc ttcggcgcga
     7501 ttttgccggt tactgcgctg taccaaatgc gggacaacgt aagcactaca tttcgctcat
     7561 cgccagccca gtcgggcggc gagttccata gcgttaaggt ttcatttagc gcctcaaata
     7621 gatcctgttc aggaaccgga tcaaagagtt cctccgccgc tggacctacc aaggcaacgc
     7681 tatgttctct tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg gctggctcga
     7741 agatacctgc aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg
     7801 gataacgcca cggaatgatg tcgtcgtgca taacaatggt gacttctaca gcgcggagaa
     7861 tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg
     7921 ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc aatatcactg tgtggcttca
     7981 ggccgccatc cactgcggag ccgtacaaat gtacggccag caacgtcggt tcgagatggc
     8041 gctcgatgac gccaactacc tctgatagtt gagtcgatac ttcggcgatc accgcttccc
     8101 tcatgatgtt taactttgtt ttagggcgac tgccctgctg cgtaacatcg ttgctgctcc
     8161 ataacatcaa acatcgaccc acggcgtaac gcgcttgctg cttggatgcc cgaggcatag
     8221 actgtacccc aaaaaaacag
//
