LOCUS       sequence-273043-        2817 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pColE1_AlphaD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2817)
  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 2817)
  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: AlphaD
FEATURES             Location/Qualifiers
     source          1..2817
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..48
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        102..120
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     102..119
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    127..183
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(185..204)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(186..204)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(211..228)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(211..227)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(220..242)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(436..455)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(592..1180)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(672..691)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1295..1397
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1398..2213
                     /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             1398..2213
                     /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(1471..1490)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2327..2782
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2414..2433)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2624..2645
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
       61 taaacccagg aggcagatca tgaccatgat tacgccaagc tatttaggtg acactataga
      121 atactcaagc ttgcatgcct gcaggtcgac tctagaggat ccccgggtac cgagctcgaa
      181 ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg
      241 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
      301 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
      361 cgaatgacgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
      421 atggtgcact ctcagtacaa tctgctctga tgccgcatag ggtctctcgc ttggctgaga
      481 cgctcgagtg acacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
      541 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
      601 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
      661 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
      721 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
      781 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
      841 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
      901 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
      961 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1021 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1081 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1141 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1201 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1261 atcaaaaagg atcttcacct agatcctttt gtgatgatcg gcacgtaaga ggttccaact
     1321 ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag
     1381 gagctaagga agctaaaatg agccatattc aacgggaaac gtcttgctca aggccgcgat
     1441 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     1501 aatcaggtgc gacaatctac cgattgtatg ggaagcccga tgcgccagag ttgtttctga
     1561 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     1621 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     1681 ggttactcac cactgcgatc ccagggaaaa cagcattcca ggtattagaa gaatatcctg
     1741 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     1801 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg actcgctcag gcgcaatcac
     1861 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     1921 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtagtca
     1981 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     2041 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     2101 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     2161 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc tgattttttt
     2221 taaggcagtt attggtgccc ttaaacgcct ggttgctacg cctgaataag tgataataag
     2281 cggatgaatg gcagaaattc gatgataagc tgtcaaacat gatgttacgc gccctgtagc
     2341 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2401 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2461 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2521 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccagc gccctgatag
     2581 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2641 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2701 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2761 aaaatattaa cgcttacaat tttaacctcg agcgtctcag cttggagaga gacccct
//
