LOCUS       sequence-201166-        3363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LI lacZ module.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3363)
  AUTHORS   Wu D, Schandry N, Lahaye T
  TITLE     A modular toolbox for Golden-Gate-based plasmid assembly streamlines
            generation of Ralstonia solanacearum species complex knockout 
            strains and multi-cassette complementation constructs.
  JOURNAL   Mol Plant Pathol. 2017 Oct 27. doi: 10.1111/mpp.12632.
  PUBMED    29077245
REFERENCE   2  (bases 1 to 3363)
  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: "Mol Plant 
            Pathol. 2017 Oct 27. doi: 10.1111/mpp.12632."
COMMENT     Sequence Label: LI LacZ
FEATURES             Location/Qualifiers
     source          1..3363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     99..118
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        99..117
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     192..211
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     405..427
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     419..436
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     420..436
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(join(989..995,996..1013,1014..1019))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1034..1055
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1175..1192)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1346..1934)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1426..1445)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2105..2896,2897..2965))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2728..2747
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2966..3070)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3138..3156
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttttaacttg gagcaggttc cattcattgt ttttttcatc atagtgaata aaatcaactg
       61 ctttaacact tgtgcctgaa caccatatcc atccggcgta atacgactca ctatagggag
      121 agcggccgcc agatcttccg gatggctcga gtttttcagc aagatttggt ctcatacgtc
      181 gtcttcctat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
      241 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
      301 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      361 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      421 taaaacgacg gccagtgaat tcgagctcgg tacctcgcga atgcatctag atatcggatc
      481 ccgggcccgt cgactgcaga ggcctgcatg caagcttggc gtaatcatgg tcatgaagac
      541 gatctgcgag accaatcttt ctagaagatc tcctacaata ttctcagctg ccatggaaaa
      601 tcgatgttct tcttttattc tctcaagatt ttcaggctgt atattaaaac ttatattaag
      661 aactatgcta accacctcat caggaaccgt tgtaggtggc gtgggttttc ttggcaatcg
      721 actctcatga aaactacgag ctaaatattc aatatgttcc tcttgaccaa ctttattctg
      781 catttttttt gaacgaggtt tagagcaagc ttcaggaaac tgagacagga attttattaa
      841 aaatttaaat tttgaagaaa gttcagggtt aatagcatcc attttttgct ttgcaagttc
      901 ctcagcattc ttaacaaaag acgtctcttt tgacatgttt aaagtttaaa cctcctgtgt
      961 gaaattatta tccgctcata attccacaca ttatacgagc cggaagcata aagtgtaaag
     1021 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgccaattg
     1081 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     1141 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     1201 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1261 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1321 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1381 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1441 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1501 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1561 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1621 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1681 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1741 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     1801 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1861 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1921 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1981 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2041 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     2101 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     2161 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     2221 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     2281 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     2341 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     2401 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     2461 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     2521 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     2581 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     2641 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     2701 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     2761 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     2821 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     2881 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     2941 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     3001 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     3061 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     3121 tgacattaac ctataaaaat aggcgtatca cgaggccgcc cctgcagccg aattatatta
     3181 tttttgccaa ataattttta acaaaagctc tgaagtcttc ttcatttaaa ttcttagatg
     3241 atacttcatc tggaaaattg tcccaattag tagcatcacg ctgtgagtaa gttctaaacc
     3301 atttttttat tgttgtatta tctctaatct tactactcga tgagttttcg gtattatctc
     3361 tat
//
