LOCUS       sequence-266875-        2472 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Engineered sgRNA-LEB2 generator including Anderson promoter J23100 
            for golden gate assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1669 to 1703)
  AUTHORS   Liu Y, Wan X, Wang B
  TITLE     Engineered CRISPRa enables programmable eukaryote-like gene 
            activation in bacteria.
  JOURNAL   Nat Commun. 2019 Aug 26;10(1):3693. doi: 10.1038/s41467-019-11479-0.
  PUBMED    31451697
REFERENCE   2  (bases 1 to 2472)
  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: "Nat 
            Commun."; date: "2019-08-26"; volume: "10(1):3693. doi"; pages: " 
            10.1038/s41467-019-11479-0"
COMMENT     Sequence Label: pLY131
FEATURES             Location/Qualifiers
     source          1..2472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(482..1297)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(482..1297)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1205..1224
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1289..1308
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1502..1520
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1524..1567)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1570..1591
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        1669..1703
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     protein_bind    1743..1761
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    1798..1816
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     misc_feature    1860..1880
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1881..1938
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(1962..1979)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(2133..2472,1..249))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2213..2232)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
       61 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      121 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      181 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      241 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
      301 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
      361 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagc
      421 tcgagtcccg tcaagtcagc gtaatgctct gccagtgtta caaccaatta accaattctg
      481 attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
      541 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
      601 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
      661 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
      721 ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt tcaacaggcc
      781 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
      841 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
      901 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
      961 attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat
     1021 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     1081 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     1141 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga
     1201 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     1261 gcctggagca agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta
     1321 tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc
     1381 agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct gagttgaagg
     1441 atcagctcga gtgccacttg acgtctaaga aaccattatt atcatgacat taacctataa
     1501 aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt tcgctaagga
     1561 tgatttctgg aattcgcggc cgcttctaga ggaagacaat agcggaaaca cagaaaaaag
     1621 cccgcacctg acagtgcggg cttttttttt cgaccaaagg tactagagtt gacggctagc
     1681 tcagtcctag gtacagtgct agctactaga atttttgaga cgttgttttg gtttgagagc
     1741 tagggccctg aagaagggcc ctagcaagtt caaataaggc tagtccgtta tcaacttggg
     1801 ccctgaagaa gggcccaagt ggcaccgagt cggtgctttt tttgaagcct attgtcttct
     1861 actagtagcg gccgctgcag tccggcaaaa aagggcaagg tgtcaccacc ctgccctttt
     1921 tctttaaaac cgaaaagatt acttcgcgtt atgcaggctt cctcgctcac tgactcgctg
     1981 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2041 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2101 aggaaccgta aaaaggccgc gttgctggcg tttttccaca ggctccgccc ccctgacgag
     2161 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2221 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2281 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2341 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2401 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2461 cacgacttat cg
//
