LOCUS       sequence-274576-        5059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human RPS5 poly(A) site reporter, stem loop mutant 1 rescue.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5059)
  AUTHORS   Wu X, Bartel DP
  TITLE     Widespread Influence of 3'-End Structures on Mammalian mRNA 
            Processing and Stability.
  JOURNAL   Cell. 2017 May 18;169(5):905-917.e11. doi: 
            10.1016/j.cell.2017.04.036.
  PUBMED    28525757
REFERENCE   2  (bases 1 to 5059)
  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.cell.2017.04"; journalName: "Cell"; date: "2017-05-18- 
            18"; volume: "169"; issue: "5"; pages: "905-917.e11"
COMMENT     Sequence Label: pCMV-GFP-RPS5pA-m1r
FEATURES             Location/Qualifiers
     source          1..5059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        318..521
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     472..492
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    544..624
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      626..635
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(632..634,635..637,638..1351)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(632..634,635..637,638..1348)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(677..698)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(938..957)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1285..1306
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2018..2037)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2083..2138
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2137..2156)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2499..2515)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2499..2515)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2512..2534)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2523..2539
                     /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(2547..2553,2554..2571,2572..2577))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2592..2613
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2672..2867
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2718..2853
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2873..3007
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2910..2929)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2964..2983
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3075..3092)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3246..3834)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3326..3345)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4005..4796,4797..4865))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4628..4647
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4866..4970)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(5001..5059,1..317)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
       61 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      121 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      181 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      241 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      301 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      361 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      421 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      481 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
      541 tccgctagcg ctaccggact cagatctcga gctcaagctt cgaattctgc agtcgacggt
      601 accgcgggcc cgggatccac cggtcgccac catggtgagc aagggcgagg agctgttcac
      661 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
      721 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
      781 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
      841 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
      901 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
      961 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     1021 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     1081 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     1141 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     1201 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     1261 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1321 cactctcggc atggacgagc tgtacaagta aagcggccgc actcctcagg tgcggctcct
     1381 cgaactccta tgccattaag aagaaggacg agctggagcg tgtggccaag tccaaccgct
     1441 gattttccca gctgctgccc aataaacctg tctggccttt gggccagtcc cagccacctg
     1501 tgctgttgtc tgtcttcggt ggggggatgt gggtggcata gccctggtgg ccttgggagc
     1561 tgctcacctt cctgacctgt tggcctgtgg ggacctggaa gtattgctga ctcagaacca
     1621 tggcacccca gggctagcag gtggggcact ggatgcaggc caggctgcct ggagtgtccc
     1681 agcaggtact ggaaatgaga tggggttatc cataacttct gagggcgtgg cacatgagat
     1741 gttgctgtgg tctgggtgga tgggtggctt gggtgcctgt gaggtctggg tagtcacaag
     1801 accatcctga gaggcttgtc accttgaagt agcactgggg agaggaggaa agctcagctg
     1861 tttagggcac acgatttcag acccctagct tcacccccct cttggagtgt ggggtggttt
     1921 cacactggcg aggttgtgct cctggttacc tccaggctgc ctatcagaag gtggtggctg
     1981 gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt
     2041 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     2101 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     2161 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     2221 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc
     2281 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
     2341 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
     2401 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga
     2461 gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     2521 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2581 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2641 cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca accatagtcc
     2701 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgccaat tctccgcccc
     2761 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     2821 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt
     2881 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2941 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     3001 ctggatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3061 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3121 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3181 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3241 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3301 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3361 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3421 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3481 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3541 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3601 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3661 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3721 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3781 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3841 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3901 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3961 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4021 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4081 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4141 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4201 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4261 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4321 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4381 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4441 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4501 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4561 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4621 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4681 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4741 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4801 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4861 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4921 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4981 cgaaaagtgc cacctgggtc gacattgatt attgactagt aatagtaatc aattacgggg
     5041 tcattagttc atagcccat
//
