LOCUS       sequence-257550-        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"
     primer_bind     complement(296..315)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             307..1122
                     /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             307..1122
                     /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(380..399)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1226..1681
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1313..1332)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1523..1544
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1717..1764
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        1818..1836
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1818..1835
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    1843..1899
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1901..1920)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1902..1920)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1927..1944)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1927..1943)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1936..1958)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2152..2171)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2308..2896)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2388..2407)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtgagctgtt ggatggggat cgcctgaatc gccccatcat ccagccagaa agtgagggag
       61 ccacggttga tgagagcttt gttgtaggtg gaccagttgg tgattttgaa cttttgcttt
      121 gccacggaac ggtctgcgtt gtcgggaaga tgcgtgatct gatccttcaa ctcagcaaaa
      181 gttcgattta ttcaacaaag ccacgttgtg tctcaaaatc tctggtgtta cattgcacaa
      241 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
      301 ggtgttatga gccatattca acgggaaacg tcttgctcaa ggccgcgatt aaattccaac
      361 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
      421 acaatctacc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
      481 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
      541 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
      601 actgcgatcc cagggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
      661 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
      721 tgtcctttta acagcgatcg cgtatttcga ctcgctcagg cgcaatcacg aatgaataac
      781 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
      841 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
      901 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
      961 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     1021 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     1081 aataaattgc agtttcattt gatgctcgat gagtttttct aatcactaga ccaatgttac
     1141 acatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     1201 atcctttttg ataatctcat gtgttacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1261 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1321 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     1381 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     1441 tagggtgatg gttcacgtag tgggccagcg ccctgataga cggtttttcg ccctttgacg
     1501 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     1561 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     1621 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     1681 ttaacagatc tcgtctcagt caggagagag acccctgttg acaattaatc atcggcatag
     1741 tatatcggca tagtataata cgacaaggtg aggaactaaa cccaggaggc agatcatgac
     1801 catgattacg ccaagctatt taggtgacac tatagaatac tcaagcttgc atgcctgcag
     1861 gtcgactcta gaggatcccc gggtaccgag ctcgaattcg ccctatagtg agtcgtatta
     1921 caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     1981 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     2041 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgacgcctga tgcggtattt
     2101 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg
     2161 ctctgatgcc gcatagggtc tctcgctaat gtgagacgag atcttgacac ggttatccac
     2221 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2281 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2341 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2401 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2461 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2521 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2581 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2641 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2701 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2761 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2821 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2881 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2941 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3001 cctttt
//
