LOCUS       sequence-230524-        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      503..958
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(590..609)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     800..821
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    979..1027
                     /note="synthetic polyadenylation signal"
     misc_feature    1041..1132
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1081..1100
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     rep_origin      complement(1256..1844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1336..1355)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1959..2061
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(2062..2488,1..389)
                     /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(2062..2488,1..389)
                     /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(2135..2154)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gcgatcgcgt atttcgactc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
       61 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
      121 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
      181 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
      241 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
      301 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
      361 ttcatttgat gctcgatgag tttttctgat tttttttaag gcagttattg gtgcccttaa
      421 acgcctggtt gctacgcctg aataagtgat aataagcgga tgaatggcag aaattcgatg
      481 ataagctgtc aaacatgatg ttacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
      541 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
      601 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
      661 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
      721 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
      781 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
      841 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
      901 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttta
      961 acgaattcgt ctcaggagaa taaaatatct ttattttcat tacatctgtg tgttggtttt
     1021 ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa
     1081 ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct ctcgctgtca
     1141 tgagacgaat tctgacacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     1201 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1261 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1321 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1381 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1441 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1501 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1561 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     1621 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     1681 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     1741 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1801 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1861 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     1921 gattatcaaa aaggatcttc acctagatcc ttttgtgatg atcggcacgt aagaggttcc
     1981 aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt
     2041 tcaggagcta aggaagctaa aatgagccat attcaacggg aaacgtcttg ctcaaggccg
     2101 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     2161 gggcaatcag gtgcgacaat ctaccgattg tatgggaagc ccgatgcgcc agagttgttt
     2221 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     2281 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     2341 gcatggttac tcaccactgc gatcccaggg aaaacagcat tccaggtatt agaagaatat
     2401 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     2461 attcctgttt gtaattgtcc ttttaaca
//
