LOCUS       sequence-206289-        4195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Anhydrotetracycline-inducible tet promoter expressing gfpuv on BBR1 
            origin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4195)
  AUTHORS   Cook TB, Rand JM, Nurani W, Courtney DK, Liu SA, Pfleger BF
  TITLE     Genetic tools for reliable gene expression and recombineering in 
            Pseudomonas putida.
  JOURNAL   J Ind Microbiol Biotechnol. 2018 Jan 3. pii: 
            10.1007/s10295-017-2001-5. doi: 10.1007/s10295-017-2001-5.
  PUBMED    29299733
REFERENCE   2  (bases 1 to 4195)
  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: "J Ind 
            Microbiol Biotechnol. 2018 Jan 3. pii: 10.1007/s10295-017-2001-5. 
            doi: 10.1007/s10295-017-2001-5."
COMMENT     Sequence Label: pBBR2k-GFPuv
FEATURES             Location/Qualifiers
     source          1..4195
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(7..633)
                     /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     538..557
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        649..704
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    685..703
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             742..1458
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(772..800)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1395..1415
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1491..1562
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1578..1605
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1672..2334)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      2335..3106
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      complement(3140..3234)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3265..4059)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3376..3395)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3986..4005
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcttaa gacccacttt cacatttaag ttgtttttct aatccgcata tgatcaattc
       61 aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga tagcttgtcg
      121 taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag cgacttgatg
      181 ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga gtgcatataa
      241 tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt cgagagtttc
      301 atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc gatgacttag
      361 taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc tttccccttc
      421 taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg cgtcgagcaa
      481 agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta gcttctgggc
      541 gagtttacgg gttgttaaac cttcgattcc gacctcatta agcagctcta atgcgctgtt
      601 aatcacttta cttttatcta atctagacat cattaattcc taatttttgt tgacactcta
      661 tcgttgatag agttatttta ccactcccta tcagtgatag agaaaagaat tcaaaagatc
      721 ttttaagaag gagatataca tatgagtaaa ggagaagaac ttttcactgg agttgtccca
      781 attcttgttg aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt
      841 gaaggtgatg caacatacgg aaaacttacc cttaaattta tttgcactac tggaaaacta
      901 cctgttccgt ggccaacact tgtcactact ttctcttatg gtgttcaatg cttttcccgt
      961 tatccggatc acatgaaacg gcatgacttt ttcaagagtg ccatgcccga aggttatgta
     1021 caggaacgca ctatatcttt caaagatgac gggaactaca agacgcgtgc tgaagtcaag
     1081 tttgaaggtg atacccttgt taatcgtatc gagttaaaag gtattgattt taaagaagat
     1141 ggaaacattc tcggacacaa actggagtac aactataact cacacaatgt atacatcacg
     1201 gcagacaaac aaaagaatgg aatcaaagct aacttcaaaa ttcgccacaa cattgaagat
     1261 ggctccgttc aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc
     1321 cttttaccag acaaccatta cctgtccaca caatctgccc tttcgaaaga tcccaacgaa
     1381 aagcgtgacc acatggtcct tcttgagttt gtaactgctg ctgggattac acatggcatg
     1441 gatgagctct acaaataagg atccaaactc gagtaaggat ctccaggcat caaataaaac
     1501 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     1561 tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat acctaggcta
     1621 cagccgatag tctggaacag cgcacttacg ggttgctgcg caacccaagt gctaccggcg
     1681 cggcagcgtg acccgtgtcg gcggctccaa cggctcgcca tcgtccagaa aacacggctc
     1741 atcgggcatc ggcaggcgct gctgcccgcg ccgttcccat tcctccgttt cggtcaaggc
     1801 tggcaggtct ggttccatgc ccggaatgcc gggctggctg ggcggctcct cgccggggcc
     1861 ggtcggtagt tgctgctcgc ccggatacag ggtcgggatg cggcgcaggt cgccatgccc
     1921 caacagcgat tcgtcctggt cgtcgtgatc aaccaccacg gcggcactga acaccgacag
     1981 gcgcaactgg tcgcggggct ggccccacgc cacgcggtca ttgaccacgt aggccgacac
     2041 ggtgccgggg ccgttgagct tcacgacgga gatccagcgc tcggccacca agtccttgac
     2101 tgcgtattgg accgtccgca aagaacgtcc gatgagcttg gaaagtgtct tctggctgac
     2161 caccacggcg ttctggtggc ccatctgcgc cacgaggtga tgcagcagca ttgccgccgt
     2221 gggtttcctc gcaataagcc cggcccacgc ctcatgcgct ttgcgttccg tttgcaccca
     2281 gtgaccgggc ttgttcttgg cttgaatgcc gatttctctg gactgcgtgg ccatgcttat
     2341 ctccatgcgg taggggtgcc gcacggttgc ggcaccatgc gcaatcagct gcaacttttc
     2401 ggcagcgcga caacaattat gcgttgcgta aaagtggcag tcaattacag attttcttta
     2461 acctacgcaa tgagctattg cggggggtgc cgcaatgagc tgttgcgtac cccccttttt
     2521 taagttgttg atttttaagt ctttcgcatt tcgccctata tctagttctt tggtgcccaa
     2581 agaagggcac ccctgcgggg ttcccccacg ccttcggcgc ggctccccct ccggcaaaaa
     2641 gtggcccctc cggggcttgt tgatcgactg cgcggccttc ggccttgccc aaggtggcgc
     2701 tgcccccttg gaacccccgc actcgccgcc gtgaggctcg gggggcaggc gggcgggctt
     2761 cgcccttcga ctgcccccac tcgcataggc ttgggtcgtt ccaggcgcgt caaggccaag
     2821 ccgctgcgcg gtcgctgcgc gagccttgac ccgccttcca cttggtgtcc aaccggcaag
     2881 cgaagcgcgc aggccgcagg ccggaggctt ttccccagag aaaattaaaa aaattgatgg
     2941 ggcaaggccg caggccgcgc agttggagcc ggtgggtatg tggtcgaagg ctgggtagcc
     3001 ggtgggcaat ccctgtggtc aagctcgtgg gcaggcgcag cctgtccatc agcttgtcca
     3061 gcagggttgt ccacgggccg agcgaagcga gccagccggt ggccgctcgc ggccatcgtc
     3121 cacatatcct agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc
     3181 tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg
     3241 agctctcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
     3301 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
     3361 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
     3421 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
     3481 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag
     3541 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
     3601 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
     3661 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     3721 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     3781 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     3841 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga
     3901 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     3961 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     4021 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     4081 ctcttgatca gatcatgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca
     4141 gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca attcc
//
