LOCUS       sequence-207219-        4320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 24x optimized PP7 stem loops (version 3) in bacterial 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4320)
  AUTHORS   Voigt F, Zhang H, Cui XA, Triebold D, Liu AX, Eglinger J, Lee ES, 
            Chao JA, Palazzo AF
  TITLE     Single-Molecule Quantification of Translation-Dependent Association 
            of mRNAs with the Endoplasmic Reticulum.
  JOURNAL   Cell Rep. 2017 Dec 26;21(13):3740-3753. doi: 
            10.1016/j.celrep.2017.12.008.
  PUBMED    29281824
REFERENCE   2  (bases 1 to 4320)
  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"; doi: 
            "10.1016/j.celrep.2017.12"; journalName: "Cell Rep"; date: 
            "2017-12-26- 26"; volume: "21"; issue: "13"; pages: "3740-3753"
COMMENT     Sequence Label: pUC57-PP7v3
FEATURES             Location/Qualifiers
     source          1..4320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     179..201
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     193..210
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     194..210
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             232..288
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1873..1938
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1959..1975)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1959..1975)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1972..1994)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1983..1999
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2007..2013,2014..2031,2032..2037))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2052..2073
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2190..2207)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2361..2949)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2441..2460)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3120..3935)
                     /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             complement(3120..3935)
                     /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     3843..3862
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3936..4040)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4108..4126
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4164..4186)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4286..4305
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
       61 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
      121 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
      181 cagtcacgac gttgtaaaac gacggccagt cccaagcttg ggggaagcgg agctaccaac
      241 ttcagcctgc tcaagcaggc tggcgacgtg gaggagaacc ctggacctat ttgcggccgc
      301 tttaccccaa gcaccagagg atatggcctc ggtgccagcg catccagcga cgtgctttct
      361 ctgtcaaacc tgcaagccag cagagcatat gggctcgctg gctccttcaa gaagttcagg
      421 aagggggacg aatctctcga aaggtccagt ggatatggcc acggacccga cgaagaacgc
      481 agcggaagac ccagcgaggt cgaccatccc accacaccat atggggtggg tgggaccaca
      541 gtcattacac tggatctcat attcagcaat cagaccgaca gtgcatatgg gcacgtcggc
      601 aacgatcaac tgctttacga caacaggcgg cggtcacagt gcagcacagc atatgggctg
      661 gctgcacccg cccagtggct ggacagagaa gaggccccca aacaccaccc agccagacca
      721 tatggggtcg gctggacatt tcccaaacca aatctccacc agaaaaaggt gggctcctgg
      781 acaggatatg gcctgccagg aagttattcc ctgctcgtcg aatatttctc aactctgcaa
      841 cggtgcagtc catatgggga cgcaccattc aagaaattca ggaaggggga cgagagtctc
      901 gaagatgcca ggacaccata tggggtgcct ggcaggagtg gaaggggctc cgaggtcgac
      961 aacgaccagc ttacgcagag tggatatggc cacctgcgaa agcattttcc caaacccaat
     1021 gtccacaaca agaaggtcac ctccacacga tatggcgtgg gaggtaccgc cagcagcgac
     1081 gtgctcagtc tgtcaaacgc catccctgcc accagatatg gctggggcag gcgatcaggc
     1141 cggcccagcg aagtcgacaa cgaccaactg ggcagcaggt catatgggac cgctgccttt
     1201 gccaagttcc gcaaggggga cgagagtctc gaggacgacg gcactccata tggggaggcc
     1261 gtctctgcct caagcgatgt tctctctctt tcaaaccttc agtccgacag aggatatggc
     1321 ctcgtcggaa ctggcaccac atcaagggat acaccaggac caagaaatcg gaccacagca
     1381 tatgggctgg gtcccgctca caacgaccac cgctatgtgt ggagaaaaaa aagtcaggcc
     1441 agagcatatg ggctcggcct gtccaatacc atccctaccg tcaagcatgg cgggggctcc
     1501 atctggagga gcacatatgg gtgccctcca accccccctg ggcgcaatct gtgctgtcga
     1561 atcggcacag cctgcagacc atatggggtc gcaggcggcc acaacgatca tcgatacgtg
     1621 tggcgcaaga aaggggcacg caccagatat ggctgggcgt gcagtgccag cagcgacgta
     1681 ctcagcctgt ccaacctcga aaccagcagg tcatatggga ccgctggttc cgaagcctgc
     1741 aaaccaaaga ataccattcc gacggtcccc tggactgcat atgggcagcc aggcaagcat
     1801 gggggagggt ctatcagatc tggaaggccc agcgagccca caggatatgg cctggggctc
     1861 ggctccggat ccgactacaa agaccacgac ggcgattaca aagatcacga tatcgattac
     1921 aaggacgacg acgacaaggg ctagccggct cgagcatggt catagctgtt tcctgtgtga
     1981 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2041 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2101 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2161 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2221 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2281 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2341 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2401 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2461 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2521 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2581 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2641 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2701 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2761 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2821 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2881 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2941 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3001 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3061 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3121 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
     3181 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     3241 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     3301 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     3361 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
     3421 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
     3481 gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa
     3541 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat
     3601 tcttctaata cctggaatgc tgttttccca gggatcgcag tggtgagtaa ccatgcatca
     3661 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt
     3721 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
     3781 aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca
     3841 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
     3901 ctagagcaag acgtttcccg ttgaatatgg ctcatactct tcctttttca atattattga
     3961 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4021 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     4081 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     4141 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     4201 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     4261 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
//
