LOCUS       sequence-273667-        4713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human RPS5 poly(A) site reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4713)
  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 4713)
  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
FEATURES             Location/Qualifiers
     source          1..4713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..588
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     539..559
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    611..691
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      693..702
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(699..701,702..704,705..1418)
                     /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(699..701,702..704,705..1415)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(744..765)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1005..1024)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1352..1373
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1739..1758)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1804..1859
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1858..1877)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2220..2236)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2220..2236)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2233..2255)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2244..2260
                     /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(2268..2274,2275..2292,2293..2298))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2313..2334
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2393..2588
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2439..2574
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2594..2728
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2631..2650)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2685..2704
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2796..2813)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2967..3555)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3047..3066)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3726..4517,4518..4586))
                     /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     4349..4368
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4587..4691)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctgggtcga cattgattat tgactagtaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      421 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      481 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      541 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
      601 cgtcagatcc gctagcgcta ccggactcag atctcgagct caagcttcga attctgcagt
      661 cgacggtacc gcgggcccgg gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc
      721 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      781 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
      841 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
      901 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      961 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1021 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1081 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1141 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     1201 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1261 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     1321 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1381 ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcact cctcaggtgc
     1441 agggctcctc gaactcctat gccattaaga agaaggacga gctggagcgt gtggccaagt
     1501 ccaaccgctg attttcccag ctgctgccca ataaacctgt ctgccctttg gggcagtccc
     1561 agccacctgt gctgttgtct gtcttcggtg gggggatgtg ggtggcatag ccctggtggc
     1621 cttgggagct gctcaccttc ctgacctgtt ggcctgtggg gacctggaag tattaggctg
     1681 cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc
     1741 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     1801 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     1861 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     1921 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
     1981 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     2041 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     2101 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     2161 ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg
     2221 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     2281 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2341 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
     2401 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2461 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2521 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2581 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2641 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     2701 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
     2761 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2821 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2881 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2941 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3001 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3061 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3121 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3181 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3241 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3301 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3361 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3421 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3481 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3541 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3601 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3661 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3721 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3781 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3841 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3901 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3961 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4021 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4081 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4141 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4201 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4261 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4321 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4381 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4441 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4501 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4561 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4621 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4681 ggggttccgc gcacatttcc ccgaaaagtg cca
//
