LOCUS       sequence-240500-        2488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcription Blocker - Synthetic polyadenylation signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2488)
  AUTHORS   Sarrion-Perdigones A, Chang L, Gonzalez Y, Gallego-Flores T, Young 
            DW, Venken KJT
  TITLE     Examining multiple cellular pathways at once using multiplex 
            hextuple luciferase assaying.
  JOURNAL   Nat Commun. 2019 Dec 13;10(1):5710. doi: 10.1038/s41467-019-13651-y.
  PUBMED    31836712
REFERENCE   2  (bases 1 to 2488)
  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: "Nat 
            Commun."; date: "2019-12-13"; volume: "10(1):5710. doi"; pages: " 
            10.1038/s41467-019-13651-y"
COMMENT     Sequence Label: p(A)n-PAUSE
FEATURES             Location/Qualifiers
     source          1..2488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      566..1021
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(653..672)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     863..884
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1042..1090
                     /note="synthetic polyadenylation signal"
     misc_feature    1104..1195
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1144..1163
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     rep_origin      complement(1319..1907)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1399..1418)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2022..2124
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(2125..2488,1..452)
                     /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(2125..2488,1..452)
                     /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(2198..2217)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
       61 acagcgatcg cgtatttcga ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
      121 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
      181 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
      241 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
      301 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
      361 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
      421 agtttcattt gatgctcgat gagtttttct gatttttttt aaggcagtta ttggtgccct
      481 taaacgcctg gttgctacgc ctgaataagt gataataagc ggatgaatgg cagaaattcg
      541 atgataagct gtcaaacatg atgttacgcg ccctgtagcg gcgcattaag cgcggcgggt
      601 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
      661 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
      721 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
      781 tagggtgatg gttcacgtag tgggccagcg ccctgataga cggtttttcg ccctttgacg
      841 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
      901 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
      961 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     1021 ttaacgaatt cgtctcagga gaataaaata tctttatttt cattacatct gtgtgttggt
     1081 tttttgtgtg aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac
     1141 aaactagcaa aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctcgctg
     1201 tcatgagacg aattctgaca cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1261 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1321 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1381 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1441 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1501 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1561 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1621 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1681 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1741 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     1801 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1861 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1921 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     1981 tgagattatc aaaaaggatc ttcacctaga tccttttgtg atgatcggca cgtaagaggt
     2041 tccaactttc accataatga aataagatca ctaccgggcg tattttttga gttatcgaga
     2101 ttttcaggag ctaaggaagc taaaatgagc catattcaac gggaaacgtc ttgctcaagg
     2161 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     2221 gtcgggcaat caggtgcgac aatctaccga ttgtatggga agcccgatgc gccagagttg
     2281 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     2341 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     2401 gatgcatggt tactcaccac tgcgatccca gggaaaacag cattccaggt attagaagaa
     2461 tatcctgatt caggtgaaaa tattgttg
//
