LOCUS       sequence-274581-        5045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  human RPS5 poly(A) site reporter, with stem loop deleted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5045)
  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 5045)
  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-del
FEATURES             Location/Qualifiers
     source          1..5045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        343..546
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     497..517
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    569..649
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      651..660
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(657..659,660..662,663..1376)
                     /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(657..659,660..662,663..1373)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(702..723)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(963..982)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1310..1331
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2029..2048)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2094..2149
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2148..2167)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2510..2526)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2510..2526)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2523..2545)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2534..2550
                     /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(2558..2564,2565..2582,2583..2588))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2603..2624
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2683..2878
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2729..2864
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2884..3018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2921..2940)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2975..2994
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3086..3103)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3257..3845)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3337..3356)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4016..4807,4808..4876))
                     /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     4639..4658
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4877..4981)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(5012..5045,1..342)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
       61 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      121 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      181 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      241 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      301 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      361 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      421 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      481 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      541 agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggactcagat ctcgagctca
      601 agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccaccggtc gccaccatgg
      661 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
      721 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
      781 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
      841 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
      901 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
      961 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1021 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1081 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     1141 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     1201 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1261 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1321 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
     1381 gccgcactcc tcaggtgcgg ctcctcgaac tcctatgcca ttaagaagaa ggacgagctg
     1441 gagcgtgtgg ccaagtccaa ccgctgattt tcccagctgc tgcccaataa acctgtcgtc
     1501 ccagccacct gtgctgttgt ctgtcttcgg tggggggatg tgggtggcat agccctggtg
     1561 gccttgggag ctgctcacct tcctgacctg ttggcctgtg gggacctgga agtattgctg
     1621 actcagaacc atggcacccc agggctagca ggtggggcac tggatgcagg ccaggctgcc
     1681 tggagtgtcc cagcaggtac tggaaatgag atggggttat ccataacttc tgagggcgtg
     1741 gcacatgaga tgttgctgtg gtctgggtgg atgggtggct tgggtgcctg tgaggtctgg
     1801 gtagtcacaa gaccatcctg agaggcttgt caccttgaag tagcactggg gagaggagga
     1861 aagctcagct gtttagggca cacgatttca gacccctagc ttcacccccc tcttggagtg
     1921 tggggtggtt tcacactggc gaggttgtgc tcctggttac ctccaggctg cctatcagaa
     1981 ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc
     2041 tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag
     2101 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac
     2161 atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac
     2221 atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat
     2281 gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat
     2341 tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca
     2401 tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct
     2461 tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt
     2521 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     2581 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2641 tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc
     2701 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccaa
     2761 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     2821 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     2881 gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2941 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3001 tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     3061 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3121 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3181 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3241 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3301 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3361 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3421 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3481 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3541 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3601 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3661 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3721 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3781 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3841 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3901 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3961 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4021 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4081 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4141 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4201 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4261 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4321 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4381 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4441 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4501 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4561 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4621 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4681 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4741 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4801 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4861 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4921 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4981 gcacatttcc ccgaaaagtg ccacctgggt cgacattgat tattgactag taatagtaat
     5041 caatt
//
