LOCUS       sequence-257563-        2811 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pColE1_AlphaE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2811)
  AUTHORS   Sarrion-Perdigones A, Gonzalez Y, Venken KJT
  TITLE     Rapid and Efficient Synthetic Assembly of Multiplex Luciferase 
            Reporter Plasmids for the Simultaneous Monitoring of Up to Six 
            Cellular Signaling Pathways.
  JOURNAL   Curr Protoc Mol Biol. 2020 Jun;131(1):e121. doi: 10.1002/cpmb.121.
  PUBMED    32539183
REFERENCE   2  (bases 1 to 2811)
  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: "Curr Protoc
            Mol Biol."; date: "2020-06"; volume: "131(1):e121. doi"; pages: " 
            10.1002/cpmb.121"
COMMENT     Sequence Label: AlphaE
FEATURES             Location/Qualifiers
     source          1..2811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..82
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        136..154
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     136..153
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    161..217
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(219..238)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(220..238)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(245..262)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(245..261)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(254..276)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(470..489)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(621..1209)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(701..720)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1324..1426
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1427..2242
                     /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             1427..2242
                     /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     complement(1500..1519)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2356..2784
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2443..2462)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2653..2674
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 taacgatatc gtctcatggc ggagagagac ccctgttgac aattaatcat cggcatagta
       61 tatcggcata gtataatacg acaaggtgag gaactaaacc caggaggcag atcatgacca
      121 tgattacgcc aagctattta ggtgacacta tagaatactc aagcttgcat gcctgcaggt
      181 cgactctaga ggatccccgg gtaccgagct cgaattcgcc ctatagtgag tcgtattaca
      241 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
      301 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
      361 atcgcccttc ccaacagttg cgcagcctga atggcgaatg acgcctgatg cggtattttc
      421 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct
      481 ctgatgccgc atagggtctc tcgcttgaga cgatatctga cacggttatc cacagaatca
      541 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      601 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      661 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      721 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      781 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      841 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      901 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      961 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1021 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     1081 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1141 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1201 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1261 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg
     1321 tgatgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg
     1381 cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatga gccatattca
     1441 acgggaaacg tcttgctcaa ggccgcgatt aaattccaac atggatgctg atttatatgg
     1501 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctacc gattgtatgg
     1561 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     1621 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     1681 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc cagggaaaac
     1741 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     1801 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     1861 cgtatttcga ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     1921 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     1981 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     2041 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     2101 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     2161 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     2221 gatgctcgat gagtttttct gatttttttt aaggcagtta ttggtgccct taaacgcctg
     2281 gttgctacgc ctgaataagt gataataagc ggatgaatgg cagaaattcg atgataagct
     2341 gtcaaacatg atgttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2401 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2461 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2521 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     2581 gttcacgtag tgggccagcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2641 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     2701 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     2761 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt t
//
