LOCUS       sequence-272181-        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"
     primer_bind     138..159
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        331..378
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        432..450
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     432..449
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    457..513
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(515..534)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(516..534)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(541..558)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(541..557)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(550..572)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(766..785)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(921..1509)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1001..1020)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1624..1726
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1727..2542
                     /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             1727..2542
                     /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(1800..1819)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(2656..2815,1..296)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2743..2762)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
       61 tgatggttca cgtagtgggc cagcgccctg atagacggtt tttcgccctt tgacgttgga
      121 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
      181 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
      241 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caattttaac
      301 ggtaccgtct caaatgggag agagacccct gttgacaatt aatcatcggc atagtatatc
      361 ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca tgaccatgat
      421 tacgccaagc tatttaggtg acactataga atactcaagc ttgcatgcct gcaggtcgac
      481 tctagaggat ccccgggtac cgagctcgaa ttcgccctat agtgagtcgt attacaattc
      541 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
      601 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
      661 cccttcccaa cagttgcgca gcctgaatgg cgaatgacgc ctgatgcggt attttctcct
      721 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
      781 tgccgcatag ggtctctcgc tgctttgaga cggtacctga cacggttatc cacagaatca
      841 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      901 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      961 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1021 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1081 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1141 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1201 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1261 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1321 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     1381 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1441 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1501 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1561 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg
     1621 tgatgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg
     1681 cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatga gccatattca
     1741 acgggaaacg tcttgctcaa ggccgcgatt aaattccaac atggatgctg atttatatgg
     1801 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctacc gattgtatgg
     1861 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     1921 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     1981 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc cagggaaaac
     2041 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     2101 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     2161 cgtatttcga ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     2221 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     2281 tttgccattc tcaccggatt cagtagtcac tcatggtgat ttctcacttg ataaccttat
     2341 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     2401 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     2461 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     2521 gatgctcgat gagtttttct gatttttttt aaggcagtta ttggtgccct taaacgcctg
     2581 gttgctacgc ctgaataagt gataataagc ggatgaatgg cagaaattcg atgataagct
     2641 gtcaaacatg atgttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2701 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2761 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggct
//
