LOCUS       sequence-206283-        4693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG-inducible trc promoter expressing gfpuv on BBR1-B5 origin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4693)
  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 4693)
  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: pBb(B5)1k-GFPuv
FEATURES             Location/Qualifiers
     source          1..4693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(166..260)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(291..1085)
                     /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(402..421)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1012..1031
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        1228..1305
                     /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             1306..2388
                     /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(1325..1344)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2620..2625,2626..2642,2643..2649)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2657..2673
                     /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             2714..3430
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2744..2772)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3367..3387
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3463..3534
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3550..3577
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 ggcttcgccc ttcgactgcc cccactcgca taggcttggg tcgttccagg cgcgtcaagg
       61 ccaagccgct gcgcggtcgc tgcgcgagcc ttgacccgcc ttccacttgg tgtccaaccg
      121 gcaagcgaag cgcgcaggcc gcaggccgga ggcactagtg cttggattct caccaataaa
      181 aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac
      241 tggatctatc aacaggagtc caagcgagct ctcgaacccc agagtcccgc tcagaagaac
      301 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
      361 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
      421 gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag
      481 cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc
      541 tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga
      601 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc
      661 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
      721 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
      781 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
      841 tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
      901 tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
      961 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
     1021 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
     1081 atcatgcgaa acgatcctca tcctgtctct tgatcagatc atgatcccct gcgccatcag
     1141 atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag
     1201 ggcgccccag ctggcaattc cgacgtcgac accatcgaat ggtgcaaaac ctttcgcggt
     1261 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
     1321 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
     1381 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
     1441 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     1501 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     1561 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     1621 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     1681 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     1741 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     1801 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     1861 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     1921 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     1981 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     2041 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     2101 gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     2161 ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     2221 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     2281 agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     2341 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     2401 taatgtaagt tagcgcgaat tgatctggtt tgacagctta tcatcgactg cacggtgcac
     2461 caatgcttct ggcgtcaggc agccatcgga agctgtggta tggctgtgca ggtcgtaaat
     2521 cactgcataa ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg
     2581 acatcataac ggttctggca aatattctga aatgagctgt tgacaattaa tcatccggct
     2641 cgtataatgt gtggaattgt gagcggataa caatttcaga attcaaaaga tcttttaaga
     2701 aggagatata catatgagta aaggagaaga acttttcact ggagttgtcc caattcttgt
     2761 tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga
     2821 tgcaacatac ggaaaactta cccttaaatt tatttgcact actggaaaac tacctgttcc
     2881 gtggccaaca cttgtcacta ctttctctta tggtgttcaa tgcttttccc gttatccgga
     2941 tcacatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaacg
     3001 cactatatct ttcaaagatg acgggaacta caagacgcgt gctgaagtca agtttgaagg
     3061 tgataccctt gttaatcgta tcgagttaaa aggtattgat tttaaagaag atggaaacat
     3121 tctcggacac aaactggagt acaactataa ctcacacaat gtatacatca cggcagacaa
     3181 acaaaagaat ggaatcaaag ctaacttcaa aattcgccac aacattgaag atggctccgt
     3241 tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc
     3301 agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg aaaagcgtga
     3361 ccacatggtc cttcttgagt ttgtaactgc tgctgggatt acacatggca tggatgagct
     3421 ctacaaataa ggatccaaac tcgagtaagg atctccaggc atcaaataaa acgaaaggct
     3481 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag
     3541 agtcacactg gctcaccttc gggtgggcct ttctgcgttt atacctaggc tacagccgat
     3601 agtctggaac agcgcactta cgggttgctg cgcaacccaa gtgctaccgg cgcggcagcg
     3661 tgacccgtgt cggcggctcc aacggctcgc catcgtccag aaaacacggc tcatcgggca
     3721 tcggcaggcg ctgctgcccg cgccgttccc attcctccgt ttcggtcaag gctggctggg
     3781 cggctcctcg ccggggccgg tcggtagttg ctgctcgccc ggatacaggg tcgggatgcg
     3841 gcgcaggtcg ccatgcccca acagcgattc gtcctggtcg tcgtgatcaa ccaccacggc
     3901 ggcactgaac accgacaggc gcaactggtc gcggggctgg ccccacgcca cgcggtcatt
     3961 gaccacgtag gccaacacgg tgccggggcc gttgagcttc acgacggaga tccagcgctc
     4021 ggccaccaag tccttgactg cgtattggac cgtccgcaaa gaacgtccga tgagcttgga
     4081 aagtgtcttc tggctgacca ccacggcgtt ctggtggccc atctgcgcca cgaggtgatg
     4141 cagcagcatt gccgccgtgg gtttcctcgc aataagcccg gcccacgcct catgcgcttt
     4201 gcgttccgtt tgcacccagt gaccgggctt gttcttggct tgaatgccga tttctctgga
     4261 ctgcgtggcc atgcttatct ccatgcggta ggggtgccgc acggttgcgg caccatgcgc
     4321 aatcagctgc aacttttcgg cagcgcgaca acaattatgc gttgcgtaaa agtggcagtc
     4381 aattacagat tttctttaac ctacgcaatg agctattgcg gggggtgccg caatgagctg
     4441 ttgcgtaccc ccctttttta agttgttgat ttttaagtct ttcgcatttc gccctatatc
     4501 tagttctttg gtgcccaaag aagggcaccc ctgcggggtt cccccacgcc ttcggcgcgg
     4561 ctccccctcc ggcaaaaagt ggcccctccg gggcttgttg atcgactgcg cggccttcgg
     4621 ccttgcccaa ggtggcgctg cccccttgga acccccgcac tcgccgccgt gaggctcggg
     4681 gggcaggcgg gcg
//
