LOCUS       sequence-206286-        5057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBBR6k-GFPuv with Anderson promoter J23109 expressing lacI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5057)
  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 5057)
  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: pBBR6k-J23109-GFPuv
FEATURES             Location/Qualifiers
     source          1..5057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(119..146)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(162..233)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(266..982)
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(309..329)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     924..952
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     protein_bind    1050..1066
                     /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)."
     protein_bind    1073..1089
                     /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)."
     primer_bind     complement(1157..1176)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1394..1428
                     /label=BBa_J23109
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.04"
     CDS             1455..2537
                     /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(1474..1493)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2722..3516
                     /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(2776..2795)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3386..3405
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      3547..3641
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3675..4446
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             join(4447..5057,1..52)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
ORIGIN
        1 cgatggcgag ccgttggagc cgccgacacg ggtcacgctg ccgcgccggt agcacttggg
       61 ttgcgcagca acccgtaagt gcgctgttcc agactatcgg ctgtagccta ggtataaacg
      121 cagaaaggcc cacccgaagg tgagccagtg tgactctagt agagagcgtt caccgacaaa
      181 caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctg
      241 gagatcctta ctcgagtttg gatccttatt tgtagagctc atccatgcca tgtgtaatcc
      301 cagcagcagt tacaaactca agaaggacca tgtggtcacg cttttcgttg ggatctttcg
      361 aaagggcaga ttgtgtggac aggtaatggt tgtctggtaa aaggacaggg ccatcgccaa
      421 ttggagtatt ttgttgataa tggtctgcta gttgaacgga gccatcttca atgttgtggc
      481 gaattttgaa gttagctttg attccattct tttgtttgtc tgccgtgatg tatacattgt
      541 gtgagttata gttgtactcc agtttgtgtc cgagaatgtt tccatcttct ttaaaatcaa
      601 taccttttaa ctcgatacga ttaacaaggg tatcaccttc aaacttgact tcagcacgcg
      661 tcttgtagtt cccgtcatct ttgaaagata tagtgcgttc ctgtacataa ccttcgggca
      721 tggcactctt gaaaaagtca tgccgtttca tgtgatccgg ataacgggaa aagcattgaa
      781 caccataaga gaaagtagtg acaagtgttg gccacggaac aggtagtttt ccagtagtgc
      841 aaataaattt aagggtaagt tttccgtatg ttgcatcacc ttcaccctct ccactgacag
      901 aaaatttgtg cccattaaca tcaccatcta attcaacaag aattgggaca actccagtga
      961 aaagttcttc tcctttactc atatgtatat ctccttctta aaagatcttt tgaattcggt
     1021 cagtgcgtcc tgctgatgtg ctcagtatct tgttatccgc tcacaatgtc aattgttatc
     1081 cgctcacaat tctcgatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca
     1141 ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac
     1201 ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga
     1261 ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc
     1321 aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgagatccc
     1381 ggacaccatc gaatttacag ctagctcagt cctagggact gtgctagcaa gagagtcaat
     1441 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
     1501 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
     1561 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
     1621 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
     1681 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
     1741 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
     1801 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
     1861 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
     1921 tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
     1981 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
     2041 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
     2101 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
     2161 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
     2221 ccattaccga gtccgggctg cgcgttggtg cggacatctc ggtagtggga tacgacgata
     2281 ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
     2341 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
     2401 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
     2461 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     2521 tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca ttaggcaccg
     2581 acgtcggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac
     2641 tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcatgatc tgatcaagag
     2701 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     2761 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     2821 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     2881 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     2941 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3001 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3061 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3121 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3181 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3241 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3301 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3361 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3421 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     3481 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgagagct
     3541 cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca
     3601 gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagcact aggatatgtg
     3661 gacgatggcc gcgagcggcc accggctggc tcgcttcgct cggcccgtgg acaaccctgc
     3721 tggacaagct gatggacagg ctgcgcctgc ccacgagctt gaccacaggg attgcccacc
     3781 ggctacccag ccttcgacca catacccacc ggctccaact gcgcggcctg cggccttgcc
     3841 ccatcaattt ttttaatttt ctctggggaa aagcctccgg cctgcggcct gcgcgcttcg
     3901 cttgccggtt ggacaccaag tggaaggcgg gtcaaggctc gcgcagcgac cgcgcagcgg
     3961 cttggccttg acgcgcctgg aacgacccaa gcctatgcga gtgggggcag tcgaagggcg
     4021 aagcccgccc gcctgccccc cgagcctcac ggcggcgagt gcgggggttc caagggggca
     4081 gcgccacctt gggcaaggcc gaaggccgcg cagtcgatca acaagccccg gaggggccac
     4141 tttttgccgg agggggagcc gcgccgaagg cgtgggggaa ccccgcaggg gtgcccttct
     4201 ttgggcacca aagaactaga tatagggcga aatgcgaaag acttaaaaat caacaactta
     4261 aaaaaggggg gtacgcaaca gctcattgcg gcaccccccg caatagctca ttgcgtaggt
     4321 taaagaaaat ctgtaattga ctgccacttt tacgcaacgc ataattgttg tcgcgctgcc
     4381 gaaaagttgc agctgattgc gcatggtgcc gcaaccgtgc ggcaccccta ccgcatggag
     4441 ataagcatgg ccacgcagtc cagagaaatc ggcattcaag ccaagaacaa gcccggtcac
     4501 tgggtgcaaa cggaacgcaa agcgcatgag gcgtgggccg ggcttattgc gaggaaaccc
     4561 acggcggcaa tgctgctgca tcacctcgtg gcgcagatgg gccaccagaa cgccgtggtg
     4621 gtcagccaga agacactttc caagctcatc ggacgttctt tgcggacggt ccaatacgca
     4681 gtcaaggact tggtggccga gcgctggatc tccgtcgtga agctcaacgg ccccggcacc
     4741 gtgtcggcct acgtggtcaa tgaccgcgtg gcgtggggcc agccccgcga ccagttgcgc
     4801 ctgtcggtgt tcagtgccgc cgtggtggtt gatcacgacg accaggacga atcgctgttg
     4861 gggcatggcg acctgcgccg catcccgacc ctgtatccgg gcgagcagca actaccgacc
     4921 ggccccggcg aggagccgcc cagccagccc ggcattccgg gcatggaacc agacctgcca
     4981 gccttgaccg aaacggagga atgggaacgg cgcgggcagc agcgcctgcc gatgcccgat
     5041 gagccgtgtt ttctgga
//
