LOCUS       sequence-206271-        4946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG-inducible trc promoter expressing gfpuv on BBR1 origin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4946)
  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 4946)
  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: pBBR1k-GFPuv
FEATURES             Location/Qualifiers
     source          1..4946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..84
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             85..1167
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(104..123)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1399..1404,1405..1421,1422..1428)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1436..1452
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             1493..2209
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1523..1551)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2146..2166
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2242..2313
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2329..2356
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2423..3085)
                     /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      3086..3857
                     /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(3891..3985)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4016..4810)
                     /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(4127..4146)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4737..4756
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcgaca ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa
       61 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
      121 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
      181 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
      241 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
      301 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
      361 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
      421 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
      481 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
      541 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
      601 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
      661 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
      721 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
      781 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
      841 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
      901 tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat
      961 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     1021 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     1081 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     1141 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agcgcgaatt
     1201 gatctggttt gacagcttat catcgactgc acggtgcacc aatgcttctg gcgtcaggca
     1261 gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct
     1321 caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa
     1381 atattctgaa atgagctgtt gacaattaat catccggctc gtataatgtg tggaattgtg
     1441 agcggataac aatttcagaa ttcaaaagat cttttaagaa ggagatatac atatgagtaa
     1501 aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
     1561 tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac
     1621 ccttaaattt atttgcacta ctggaaaact acctgttccg tggccaacac ttgtcactac
     1681 tttctcttat ggtgttcaat gcttttcccg ttatccggat cacatgaaac ggcatgactt
     1741 tttcaagagt gccatgcccg aaggttatgt acaggaacgc actatatctt tcaaagatga
     1801 cgggaactac aagacgcgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatcgtat
     1861 cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt ctcggacaca aactggagta
     1921 caactataac tcacacaatg tatacatcac ggcagacaaa caaaagaatg gaatcaaagc
     1981 taacttcaaa attcgccaca acattgaaga tggctccgtt caactagcag accattatca
     2041 acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac
     2101 acaatctgcc ctttcgaaag atcccaacga aaagcgtgac cacatggtcc ttcttgagtt
     2161 tgtaactgct gctgggatta cacatggcat ggatgagctc tacaaataag gatccaaact
     2221 cgagtaagga tctccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     2281 cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg
     2341 ggtgggcctt tctgcgttta tacctaggct acagccgata gtctggaaca gcgcacttac
     2401 gggttgctgc gcaacccaag tgctaccggc gcggcagcgt gacccgtgtc ggcggctcca
     2461 acggctcgcc atcgtccaga aaacacggct catcgggcat cggcaggcgc tgctgcccgc
     2521 gccgttccca ttcctccgtt tcggtcaagg ctggcaggtc tggttccatg cccggaatgc
     2581 cgggctggct gggcggctcc tcgccggggc cggtcggtag ttgctgctcg cccggataca
     2641 gggtcgggat gcggcgcagg tcgccatgcc ccaacagcga ttcgtcctgg tcgtcgtgat
     2701 caaccaccac ggcggcactg aacaccgaca ggcgcaactg gtcgcggggc tggccccacg
     2761 ccacgcggtc attgaccacg taggccgaca cggtgccggg gccgttgagc ttcacgacgg
     2821 agatccagcg ctcggccacc aagtccttga ctgcgtattg gaccgtccgc aaagaacgtc
     2881 cgatgagctt ggaaagtgtc ttctggctga ccaccacggc gttctggtgg cccatctgcg
     2941 ccacgaggtg atgcagcagc attgccgccg tgggtttcct cgcaataagc ccggcccacg
     3001 cctcatgcgc tttgcgttcc gtttgcaccc agtgaccggg cttgttcttg gcttgaatgc
     3061 cgatttctct ggactgcgtg gccatgctta tctccatgcg gtaggggtgc cgcacggttg
     3121 cggcaccatg cgcaatcagc tgcaactttt cggcagcgcg acaacaatta tgcgttgcgt
     3181 aaaagtggca gtcaattaca gattttcttt aacctacgca atgagctatt gcggggggtg
     3241 ccgcaatgag ctgttgcgta cccccctttt ttaagttgtt gatttttaag tctttcgcat
     3301 ttcgccctat atctagttct ttggtgccca aagaagggca cccctgcggg gttcccccac
     3361 gccttcggcg cggctccccc tccggcaaaa agtggcccct ccggggcttg ttgatcgact
     3421 gcgcggcctt cggccttgcc caaggtggcg ctgccccctt ggaacccccg cactcgccgc
     3481 cgtgaggctc ggggggcagg cgggcgggct tcgcccttcg actgccccca ctcgcatagg
     3541 cttgggtcgt tccaggcgcg tcaaggccaa gccgctgcgc ggtcgctgcg cgagccttga
     3601 cccgccttcc acttggtgtc caaccggcaa gcgaagcgcg caggccgcag gccggaggct
     3661 tttccccaga gaaaattaaa aaaattgatg gggcaaggcc gcaggccgcg cagttggagc
     3721 cggtgggtat gtggtcgaag gctgggtagc cggtgggcaa tccctgtggt caagctcgtg
     3781 ggcaggcgca gcctgtccat cagcttgtcc agcagggttg tccacgggcc gagcgaagcg
     3841 agccagccgg tggccgctcg cggccatcgt ccacatatcc tagtgcttgg attctcacca
     3901 ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
     3961 attactggat ctatcaacag gagtccaagc gagctctcga accccagagt cccgctcaga
     4021 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt
     4081 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag
     4141 ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag
     4201 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga
     4261 gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc
     4321 cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg
     4381 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat
     4441 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg
     4501 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga
     4561 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg
     4621 cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc
     4681 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc
     4741 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt
     4801 gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agatcatgat cccctgcgcc
     4861 atcagatcct tggcggcaag aaagccatcc agtttacttt gcagggcttc ccaaccttac
     4921 cagagggcgc cccagctggc aattcc
//
