LOCUS       sequence-274574-        5059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human RPS5 poly(A) site reporter, stem loop mutant 1.
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-m1
FEATURES             Location/Qualifiers
     source          1..5059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        35..410
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        411..614
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     565..585
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    637..717
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      719..728
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(725..727,728..730,731..1444)
                     /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(725..727,728..730,731..1441)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(770..791)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1031..1050)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1378..1399
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2111..2130)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2176..2231
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2230..2249)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2592..2608)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2592..2608)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2605..2627)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2616..2632
                     /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(2640..2646,2647..2664,2665..2670))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2685..2706
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2765..2960
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2811..2946
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2966..3100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3003..3022)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3057..3076
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3168..3185)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3339..3927)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3419..3438)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4098..4889,4890..4958))
                     /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     4721..4740
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4959..5059,1..4))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgcgcacatt tccccgaaaa gtgccacctg ggtcgacatt gattattgac tagtaatagt
       61 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      121 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      181 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      241 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      301 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      361 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      421 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      481 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      541 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      601 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct
      661 cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggtcgc
      721 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      781 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      841 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      901 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      961 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1021 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1081 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1141 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1201 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1261 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1321 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1381 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1441 gtaaagcggc cgcactcctc aggtgcggct cctcgaactc ctatgccatt aagaagaagg
     1501 acgagctgga gcgtgtggcc aagtccaacc gctgattttc ccagctgctg cccaataaac
     1561 ctgtctgccc tttgggccag tcccagccac ctgtgctgtt gtctgtcttc ggtgggggga
     1621 tgtgggtggc atagccctgg tggccttggg agctgctcac cttcctgacc tgttggcctg
     1681 tggggacctg gaagtattgc tgactcagaa ccatggcacc ccagggctag caggtggggc
     1741 actggatgca ggccaggctg cctggagtgt cccagcaggt actggaaatg agatgtggtt
     1801 atccataact tctgagggcg tggcacatga gatgttgctg tggtctgggt ggatgggtgg
     1861 cttgggtgcc tgtgaggtct gggtagtcac aagaccatcc tgagaggctt gtcaccttga
     1921 agtagcactg gggagaggag gaaagctcag ctgtttaggg cacacgattt cagaccccta
     1981 gcttcacccc cctcttggag tgtggggtgg tttcacactg gcgaggttgt gctcctggtt
     2041 acctccaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat
     2101 accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     2161 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     2221 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     2281 atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc
     2341 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa
     2401 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa
     2461 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac
     2521 tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag cccaagcttg
     2581 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     2641 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     2701 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg
     2761 atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2821 taactccgcc cagttccgcc aattctccgc cccatggctg actaattttt tttatttatg
     2881 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2941 gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata
     3001 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3061 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc
     3121 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3181 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3241 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3301 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3361 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3421 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3481 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3541 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3601 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3661 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3721 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3781 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3841 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3901 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3961 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4021 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4081 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4141 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4201 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4261 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4321 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4381 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4441 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4501 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4561 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4621 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4681 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4741 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4801 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4861 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4921 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4981 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5041 aataaacaaa taggggttc
//
