LOCUS       sequence-201154-        2762 bp    DNA     circular SYN 15-APR-2025
DEFINITION  LI promoter module.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2762)
  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 2762)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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 A-B pEPS
FEATURES             Location/Qualifiers
     source          1..2762
                     /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(415..431)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(415..431)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(428..450)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    439..455
                     /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(463..469,470..487,488..493))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    508..529
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(646..663)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(817..1405)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(897..916)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1520..1525,1526..1542,1543..1548)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1737..2270
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     2357..2375
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2413..2435)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2535..2554
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(2748..2762,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(2762,1..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 aggtacgggt ctcagcggct tcttcacgcg cgagaagccc
      121 gcgaggggcg gcgcgtggca cgggctgcaa gctcctgcgt gccgtccgtt cagtctgact
      181 tccgtaagcc agacttttaa gaaatcgaat tctgtatcga attgacgctg cgcgtgcgaa
      241 taacgcaccc aactgtaaaa cgtatccact ccgaagtagg gaaacgaaat gagcataaag
      301 ccgcctgttg gtattgaagc gtcaccaatt gcggccatac actttgccac tcccgacgaa
      361 gcggttgtgc aatctgcgag acctctgcct gcatgcaagc ttggcgtaat catggtcata
      421 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
      481 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
      541 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
      601 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
      661 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      721 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      781 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      841 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      901 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      961 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1021 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1081 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1141 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1201 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1261 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1321 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1381 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1441 tcagtggaac gaaaactcac gttaagggat tttggtcatg aggcgcacac cgtggaaacg
     1501 gatgaaggca cgaacccagt tgacataagc ctgttcggtt cgtaaactgt aatgcaagta
     1561 gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc
     1621 agtggcggtt ttcatggctt gttatgactg tttttttgta cagtctatgc ctcgggcatc
     1681 caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt
     1741 tacgcagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaggtggct
     1801 caagtatggg catcattcgc acatgtaggc tcggccctga ccaagtcaaa tccatgcggg
     1861 ctgctcttga tcttttcggt cgtgagttcg gcgacgtagc cacctactcc caacatcagc
     1921 cggactccga ttacctcggg aacttgctcc gtagtaagac attcatcgcg cttgctgcct
     1981 tcgaccaaga agcggttgtt ggcgctctcg cggcttacgt tctgcccaag tttgagcagc
     2041 cgcgtagtga gatctatatc tatgatctcg cagtctccgg agagcaccgg aggcagggca
     2101 ttgccaccgc gctcatcaat ctcctcaagc atgaggccaa cgcgcttggt gcttatgtga
     2161 tctacgtgca agcagattac ggtgacgatc ccgcagtggc tctctataca aagttgggca
     2221 tacgggaaga agtgatgcac tttgatatcg acccaagtac cgccacctaa caattcgttc
     2281 aagccgagat cggcttcccg gccgcggagt tgttcggtaa attgtcacaa cgccgctcat
     2341 gacattaacc tataaaaata ggcgtatcac gaggcccttt tgtctcgcgc gtttcggtga
     2401 tgacggtgaa aacctctgac acatgcagct cccggagaag gtcacagctt gtctgtaagc
     2461 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     2521 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     2581 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     2641 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     2701 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     2761 tt
//
