LOCUS       sequence-272182-        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"
     primer_bind     complement(29..48)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     239..260
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        432..479
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        533..551
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     533..550
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    558..614
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(616..635)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(617..635)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(642..659)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(642..658)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(651..673)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(867..886)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1018..1606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1098..1117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1721..1823
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1824..2639
                     /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             1824..2639
                     /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(1897..1916)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(2753..2811,1..397)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
       61 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      121 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      181 ccagcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      241 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      301 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
      361 aacgcgaatt ttaacaaaat attaacgctt acaattttaa cgatatcgtc tcatggcgga
      421 gagagacccc tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
      481 aggtgaggaa ctaaacccag gaggcagatc atgaccatga ttacgccaag ctatttaggt
      541 gacactatag aatactcaag cttgcatgcc tgcaggtcga ctctagagga tccccgggta
      601 ccgagctcga attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa
      661 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
      721 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
      781 agcctgaatg gcgaatgacg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
      841 tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata gggtctctcg
      901 cttgagacga tatctgacac ggttatccac agaatcaggg gataacgcag gaaagaacat
      961 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1021 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1081 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1141 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1201 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1261 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1321 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1381 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1441 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1501 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1561 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1621 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1681 gagattatca aaaaggatct tcacctagat ccttttgtga tgatcggcac gtaagaggtt
     1741 ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat
     1801 tttcaggagc taaggaagct aaaatgagcc atattcaacg ggaaacgtct tgctcaaggc
     1861 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     1921 tcgggcaatc aggtgcgaca atctaccgat tgtatgggaa gcccgatgcg ccagagttgt
     1981 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
     2041 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     2101 atgcatggtt actcaccact gcgatcccag ggaaaacagc attccaggta ttagaagaat
     2161 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     2221 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgactc gctcaggcgc
     2281 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     2341 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
     2401 tagtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     2461 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     2521 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     2581 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctgat
     2641 tttttttaag gcagttattg gtgcccttaa acgcctggtt gctacgcctg aataagtgat
     2701 aataagcgga tgaatggcag aaattcgatg ataagctgtc aaacatgatg ttacgcgccc
     2761 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac c
//
