LOCUS       sequence-257552-        2815 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pColE1_AlphaC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2815)
  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 2815)
  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: AlphaC
FEATURES             Location/Qualifiers
     source          1..2815
                     /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(625..1213)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(705..724)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1328..1430
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1431..2246
                     /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             1431..2246
                     /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(1504..1523)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2360..2788
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2447..2466)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2657..2678
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 taacggtacc gtctcaaatg 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 tcgctgcttt gagacggtac ctgacacggt tatccacaga
      541 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
      601 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
      661 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
      721 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
      781 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
      841 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
      901 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
      961 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1021 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     1081 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1141 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1201 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1261 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     1321 tttgtgatga tcggcacgta agaggttcca actttcacca taatgaaata agatcactac
     1381 cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa atgagccata
     1441 ttcaacggga aacgtcttgc tcaaggccgc gattaaattc caacatggat gctgatttat
     1501 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc taccgattgt
     1561 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     1621 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     1681 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atcccaggga
     1741 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     1801 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     1861 atcgcgtatt tcgactcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     1921 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     1981 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     2041 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     2101 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     2161 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     2221 atttgatgct cgatgagttt ttctgatttt ttttaaggca gttattggtg cccttaaacg
     2281 cctggttgct acgcctgaat aagtgataat aagcggatga atggcagaaa ttcgatgata
     2341 agctgtcaaa catgatgtta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2401 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2461 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2521 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2581 gatggttcac gtagtgggcc agcgccctga tagacggttt ttcgcccttt gacgttggag
     2641 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2701 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2761 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aattt
//
