LOCUS       sequence-212665-        4941 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBBR1k-GFPuv with Anderson promoter J23107 expressing lacI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4941)
  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 4941)
  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-J23107-GFPuv
FEATURES             Location/Qualifiers
     source          1..4941
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..23,24..30)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    38..54
                     /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             95..811
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(125..153)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     748..768
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      844..915
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      931..958
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1025..1687)
                     /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      1688..2459
                     /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(2493..2587)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2618..3412)
                     /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(2729..2748)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3339..3358
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        3567..3601
                     /label=BBa_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     CDS             3628..4710
                     /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(3647..3666)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttgacaatta atcatccggc tcgtataatg tgtggaattg tgagcggata acaatttcag
       61 aattcaaaag atcttttaag aaggagatat acatatgagt aaaggagaag aacttttcac
      121 tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt
      181 cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac
      241 tactggaaaa ctacctgttc cgtggccaac acttgtcact actttctctt atggtgttca
      301 atgcttttcc cgttatccgg atcacatgaa acggcatgac tttttcaaga gtgccatgcc
      361 cgaaggttat gtacaggaac gcactatatc tttcaaagat gacgggaact acaagacgcg
      421 tgctgaagtc aagtttgaag gtgataccct tgttaatcgt atcgagttaa aaggtattga
      481 ttttaaagaa gatggaaaca ttctcggaca caaactggag tacaactata actcacacaa
      541 tgtatacatc acggcagaca aacaaaagaa tggaatcaaa gctaacttca aaattcgcca
      601 caacattgaa gatggctccg ttcaactagc agaccattat caacaaaata ctccaattgg
      661 cgatggccct gtccttttac cagacaacca ttacctgtcc acacaatctg ccctttcgaa
      721 agatcccaac gaaaagcgtg accacatggt ccttcttgag tttgtaactg ctgctgggat
      781 tacacatggc atggatgagc tctacaaata aggatccaaa ctcgagtaag gatctccagg
      841 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
      901 tcggtgaacg ctctctacta gagtcacact ggctcacctt cggctgggcc tttctgcgtt
      961 tatacctagg ctacagccga tagtctggaa cagcgcactt acgggttgct gcgcaaccca
     1021 agtgctaccg gcgcggcagc gtgacccgtg tcggcggctc caacggctcg ccatcgtcca
     1081 gaaaacacgg ctcatcgggc atcggcaggc gctgctgccc gcgccgttcc cattcctccg
     1141 tttcggtcaa ggctggcagg tctggttcca tgcccggaat gccgggctgg ctgggcggct
     1201 cctcgccggg gccggtcggt agttgctgct cgcccggata cagggtcggg atgcggcgca
     1261 ggtcgccatg ccccaacagc gattcgtcct ggtcgtcgtg atcaaccacc acggcggcac
     1321 tgaacaccga caggcgcaac tggtcgcggg gctggcccca cgccacgcgg tcattgacca
     1381 cgtaggccga cacggtgccg gggccgttga gcttcacgac ggagatccag cgctcggcca
     1441 ccaagtcctt gactgcgtat tggaccgtcc gcaaagaacg tccgatgagc ttggaaagtg
     1501 tcttctggct gaccaccacg gcgttctggt ggcccatctg cgccacgagg tgatgcagca
     1561 gcattgccgc cgtgggtttc ctcgcaataa gcccggccca cgcctcatgc gctttgcgtt
     1621 ccgtttgcac ccagtgaccg ggcttgttct tggcttgaat gccgatttct ctggactgcg
     1681 tggccatgct tatctccatg cggtaggggt gccgcacggt tgcggcacca tgcgcaatca
     1741 gctgcaactt ttcggcagcg cgacaacaat tatgcgttgc gtaaaagtgg cagtcaatta
     1801 cagattttct ttaacctacg caatgagcta ttgcgggggg tgccgcaatg agctgttgcg
     1861 tacccccctt ttttaagttg ttgattttta agtctttcgc atttcgccct atatctagtt
     1921 ctttggtgcc caaagaaggg cacccctgcg gggttccccc acgccttcgg cgcggctccc
     1981 cctccggcaa aaagtggccc ctccggggct tgttgatcga ctgcgcggcc ttcggccttg
     2041 cccaaggtgg cgctgccccc ttggaacccc cgcactcgcc gccgtgaggc tcggggggca
     2101 ggcgggcggg cttcgccctt cgactgcccc cactcgcata ggcttgggtc gttccaggcg
     2161 cgtcaaggcc aagccgctgc gcggtcgctg cgcgagcctt gacccgcctt ccacttggtg
     2221 tccaaccggc aagcgaagcg cgcaggccgc aggccggagg cttttcccca gagaaaatta
     2281 aaaaaattga tggggcaagg ccgcaggccg cgcagttgga gccggtgggt atgtggtcga
     2341 aggctgggta gccggtgggc aatccctgtg gtcaagctcg tgggcaggcg cagcctgtcc
     2401 atcagcttgt ccagcagggt tgtccacggg ccgagcgaag cgagccagcc ggtggccgct
     2461 cgcggccatc gtccacatat cctagtgctt ggattctcac caataaaaaa cgcccggcgg
     2521 caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac
     2581 aggagtccaa gcgagctctc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc
     2641 gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt
     2701 cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat
     2761 agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca
     2821 ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca
     2881 tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca
     2941 gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt
     3001 tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat
     3061 cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg
     3121 gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg
     3181 cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc tgcagttcat
     3241 tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc
     3301 ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc
     3361 tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg
     3421 atcctcatcc tgtctcttga tcagatcatg atcccctgcg ccatcagatc cttggcggca
     3481 agaaagccat ccagtttact ttgcagggct tcccaacctt accagagggc gccccagctg
     3541 gcaattccga cgtcgacacc atcgaattta cggctagctc agccctaggt attatgctag
     3601 caagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
     3661 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
     3721 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
     3781 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
     3841 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
     3901 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
     3961 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
     4021 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
     4081 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
     4141 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
     4201 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
     4261 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
     4321 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
     4381 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
     4441 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag
     4501 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
     4561 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg
     4621 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     4681 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagcgcga
     4741 attgatctgg tttgacagct tatcatcgac tgcacggtgc accaatgctt ctggcgtcag
     4801 gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc
     4861 gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg
     4921 caaatattct gaaatgagct g
//
