LOCUS       sequence-201167-        4779 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LIII backbone vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4779)
  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 4779)
  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: LIII 1-6 Amp RC
FEATURES             Location/Qualifiers
     source          1..4779
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1200..1216)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1200..1216)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1213..1235)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1224..1240
                     /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)."
     promoter        complement(join(1248..1254,1255..1272,1273..1278))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1293..1314
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1431..1448)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1602..2190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1682..1701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2361..3152,3153..3221))
                     /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     2984..3003
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3222..3326)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3394..3412
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3450..3472)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4552..4571
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4765..4779,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4779,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttcgagctcg gtacctcgcg aatgcatcta gatatcggat
       61 cccgggcccg tcgactgcag aggcctgcat gcaagcttgg cgtaatcatg gtcatgaaga
      121 cgatctgtcg atgtcttggg gccagcgatt gtatcggggg atactcccca tcagctattc
      181 agaaccctca agacaactct ggacatcagc ggctgccact tgtccaaatc gcatcatggc
      241 aacgagacat tggtggttgc tatggagaga gggcttgcac gccaaaagcc agcgtcgtcc
      301 ctcactcctc aacctcgctg cgaaccatca gcacaggttt gttcgtcttg cgcactacgc
      361 cttcggccac actgccaagc accaggtgtt tgagcccgcg ccggccgtga gtaccaagca
      421 caatcaggtc agtgcccgtc tcgttggcct cggccacaat cgttgcggaa atctgccccg
      481 gcacaatgga tttctctagc aacttggtcg aatgcttgac gccagcggcc tccaatttgg
      541 tcgcaacctc ggccagcgtg tctcggccga actcgacaac ctgtttggcg taggcaacgg
      601 agtcagcccc gaccatgtcg aaggtcacgt cgctgctatc ggcaacaaag agggcttcga
      661 tttcggcgcc cgtcgctttg ctgaccacaa tcgcctggtg cagggcgaga taggacgcgt
      721 ggctgccgtc gatggcgagc aggatgcgtt ggtacatgac ggtttccaaa ggcgttgtgc
      781 gggacagtgc cgcatgttac cagcgtaggc ggcaggatgc tgggctcatt gacccatatc
      841 aacgctcttc cggggcgatt cccaataccg ccgcgcaggc aatgcgcccg gcacgcaggt
      901 tgtccagagc ggtgttggtt tcctgcagtc gatacggcat gacgtgaacc tgcaggggta
      961 ctgcagccgc cacgtgcatg agcgccaggc catccgatcg cgtcaggtta gccactgaga
     1021 cgagcttgcg ctcctcccac agcagccggt acggaaacgt ggggatgtcg ctcatggggc
     1081 tgccaccgca aaccacggaa gggtgtgaat tcgagctcgg tacctcgcga atgcatctag
     1141 atatcggatc ccgggcccgt cgactgcaga ggcctgcatg caagcttggc gtaatcatgg
     1201 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1261 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1321 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     1381 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1441 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1501 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1561 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1621 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1681 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1741 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1801 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1861 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1921 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1981 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2041 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2101 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2161 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2221 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2281 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2341 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2401 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2461 gagggcttac catctggccc cagtgctgca atgataccgc gagatccacg ctcaccggct
     2521 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2581 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2641 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     2701 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     2761 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     2821 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     2881 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2941 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3001 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3061 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3121 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3181 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3241 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3301 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     3361 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     3421 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     3481 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     3541 gttggcgggt gtcggggctg gcttaactca cctgaccgag cagatgccgg tggtgacggt
     3601 cgcgcccaac gacgcgctgc tggagaagct caagtccaac atccaggaag tgcgcgcgcg
     3661 cggcggcaag ctctacgtgt tcgccgatgc cgacacgcat atccagtcga gcgacggcat
     3721 ccaggtgatc cgcatgcccg agcactatgg gcagctgtcg ccgatcctgc acgtggtgcc
     3781 gctgcagctg ctggcgtatc acaccgccgt ggcgcgcggg acggatgtgg ataagccgcg
     3841 gaatttggct aagtctgtga cggtggagtg agggggcggt agtgctgttg tccttgtaac
     3901 ggcagacaaa aaagtctgta ccaggtccgc agttgaaacg tataggcacc ggtctgcccc
     3961 gattaagcgg ggggccggtg tttttttcga ataggtgttg gctcagagga ggcgggcgcc
     4021 gccgctgttt cccagacctt agtcgtaggg tgaccagcgg acccggtcgt agcaggccga
     4081 cggcaattcg atgctcaacg ttccgaattt atccagcagc ttcttggcga gcttgggtga
     4141 cgcgacctgt tccgcgtata aggtgactag gtgtctcacc tcatatcggc gcactttgcg
     4201 atcgcttttc cgatcgaccg actgagacga ttcaacgttt tcttcgggaa agcacgctgt
     4261 ttccgctatt tgagatgttc actggggcgc aattctctgc ccgggaagaa cggggccgca
     4321 tcgatgcggt acttcaaagc ctcccaaagc gcatggttgg cgcattcggg gggccgactc
     4381 ttgtgtccgc ttgatgccac gtctcggatc gtgagccccg gtcgatagtt ggagcctctt
     4441 ccactgccca ggccggcggc tcgccatcgt tctatatcgg caatcatgat gttgcggcgc
     4501 ttcttcataa ctgatcagct caatcctgtc gccctacgtc gtcttcctat gcggcatcag
     4561 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     4621 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     4681 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     4741 taagttgggt aacgccaggg ttttcccagt cacgacgtt
//
