LOCUS       sequence-149380-        3340 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Transposon-propagation plasmid for Mu-BsaI modified transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3340)
  AUTHORS   Nadler DC, Morgan SA, Flamholz A, Kortright KE, Savage DF
  TITLE     Rapid construction of metabolite biosensors using domain-insertion 
            profiling.
  JOURNAL   Nat Commun. 2016 Jul 29;7:12266. doi: 10.1038/ncomms12266.
  PUBMED    27470466
REFERENCE   2  (bases 1 to 3340)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2016-07-29"; volume: "7:12266. doi"; pages: " 
            10.1038/ncomms12266"
COMMENT     Sequence Label: pUCKanR-Mu-BsaI
FEATURES             Location/Qualifiers
     source          1..3340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     720..739
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     804..823
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      931..1519
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1420..1439
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1673..1690
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1807..1828
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1843..1848,1849..1866,1867..1873)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1881..1897
                     /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     1886..1908
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     promoter        2101..2203
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2204..2863
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2270..2289)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     3290..3312
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(join(3337..3340,1..812))
                     /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(join(3337..3340,1..812))
                     /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"
ORIGIN
        1 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
       61 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
      121 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
      181 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
      241 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca
      301 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
      361 tgagcgaggc gaaatacgcg atcactgtta aaaggacaat tacaaacagg aatcgaatgc
      421 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
      481 tctaatacct ggaatgctgt tttgccgggg atcgcagtgg tgagtaacca tgcatcatca
      541 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
      601 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
      661 tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta
      721 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta
      781 gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
      841 gcagacagtt ttattgttca tgaccatccc ttaacgtgag ttttcgttcc actgagcgtc
      901 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
      961 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1021 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1081 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1141 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1201 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1261 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1321 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1381 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     1441 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     1501 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     1561 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     1621 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     1681 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     1741 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     1801 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     1861 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatgt
     1921 ccagatctgc atcggagacc gaaaaacgcg aaagcgtttc acgataaatg cgaaaacgga
     1981 tcgatccttt tcgaccgaat aaatacctgt gacggaagat cacttcgcag aataaataaa
     2041 tcctggtgtc cctgttgata ccgggaagcc ctgggccaac ttttggcgaa aatgagatgt
     2101 tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt
     2161 attttttgag ttgtcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac
     2221 tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca
     2281 gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa
     2341 gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct
     2401 gatgaatgct catccggaat tacgtatggc aatgaaagac ggtgagctgg tgatatggga
     2461 tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg
     2521 gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg
     2581 ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtgtc
     2641 agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt
     2701 cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc
     2761 gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa
     2821 tgaattacaa cagtactgcg atgagtggca gggcggggcg taattttttt aaggcagtta
     2881 ttggtgccct taaacgcctg gttgctacgc ctgaataagt gataataagc ggatgaatgg
     2941 cagaaattcg aaagcaaatt cgacccggtc gtcggttcag ggcagggtcg ttaaatagcc
     3001 gcttatgtct attgctggtt taccggttta ttgactaccg gaagcagtgt gaccgtgtgc
     3061 ttctcaaatg cctgaggcca gtttgctcag gctctccccg tggaggtaat aattgacgat
     3121 aggatcgatc cgttttcgca tttatcgtga aacgctttcg cgtttttcgg tctccgcgtc
     3181 aagcttagcg gttaattaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     3241 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtcac cgggagctgc
     3301 atgtgtcaga ggttttcacc gtcatcaccg aaacgcttag
//
