LOCUS       sequence-272009-        3006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pColE1_AlphaB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3006)
  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 3006)
  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: AlphaB
FEATURES             Location/Qualifiers
     source          1..3006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      122..577
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(209..228)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     419..440
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        613..660
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        714..732
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     714..731
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    739..795
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(797..816)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(798..816)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(823..840)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(823..839)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(832..854)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1048..1067)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1204..1792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1284..1303)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2198..2217)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             join(2209..3006,1..18)
                     /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             join(2209..3006,1..18)
                     /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(2282..2301)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ctcgatgagt ttttctaatc actagaccaa tgttacacat atatacttta gattgattta
       61 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgtgt
      121 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
      181 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      241 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      301 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      361 ccagcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      421 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      481 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
      541 aacgcgaatt ttaacaaaat attaacgctt acaattttaa cagatctcgt ctcagtcagg
      601 agagagaccc ctgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac
      661 aaggtgagga actaaaccca ggaggcagat catgaccatg attacgccaa gctatttagg
      721 tgacactata gaatactcaa gcttgcatgc ctgcaggtcg actctagagg atccccgggt
      781 accgagctcg aattcgccct atagtgagtc gtattacaat tcactggccg tcgttttaca
      841 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
      901 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
      961 cagcctgaat ggcgaatgac gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     1021 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agggtctctc
     1081 gctaatgtga gacgagatct tgacacggtt atccacagaa tcaggggata acgcaggaaa
     1141 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1201 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1261 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1321 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1381 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1441 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1501 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1561 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1621 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     1681 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1741 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1801 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1861 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttgtgagctg ttggatgggg
     1921 atcgcctgaa tcgccccatc atccagccag aaagtgaggg agccacggtt gatgagagct
     1981 ttgttgtagg tggaccagtt ggtgattttg aacttttgct ttgccacgga acggtctgcg
     2041 ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa aagttcgatt tattcaacaa
     2101 agccacgttg tgtctcaaaa tctctggtgt tacattgcac aagataaaaa tatatcatca
     2161 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     2221 caacgggaaa cgtcttgctc aaggccgcga ttaaattcca acatggatgc tgatttatat
     2281 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta ccgattgtat
     2341 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     2401 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     2461 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat cccagggaaa
     2521 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     2581 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     2641 cgcgtatttc gactcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     2701 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     2761 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     2821 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     2881 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     2941 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     3001 ttgatg
//
