LOCUS       sequence-274586-        5654 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human ILVBL poly(A) site reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5654)
  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 5654)
  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-ILVBLpA
FEATURES             Location/Qualifiers
     source          1..5654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        373..576
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     527..547
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    599..679
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      681..690
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(687..689,690..692,693..1406)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNDHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(732..753)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(993..1012)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1340..1361
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2668..2687)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2733..2788
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2787..2806)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3149..3165)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3149..3165)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3162..3184)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3173..3189
                     /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(3197..3203,3204..3221,3222..3227))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3242..3263
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3322..3517
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3368..3503
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3523..3657
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3560..3579)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3614..3633
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3725..3742)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3896..4484)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3976..3995)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4655..5446,5447..5515))
                     /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     5278..5297
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5516..5620)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(5651..5654,1..372)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ttgattattg actagtaata gtaatcaatt acggggtcat tagttcatag cccatatatg
       61 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      121 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      181 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      241 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      301 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      361 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      421 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      481 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      541 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
      601 tagcgctacc ggactcagat ctcgagctca agcttcgaat tctgcagtcg acggtaccgc
      661 gggcccggga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
      721 tggtgcccat cctggtcgag ctggacggcg acgtaaacga ccacaagttc agcgtgtccg
      781 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
      841 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
      901 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
      961 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1021 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1081 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     1141 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     1201 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1261 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     1321 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1381 tcggcatgga cgagctgtac aagtaaagcg gccgcactcc tcaggtgcga tcaggtggtc
     1441 aaggtgctgc acgatgccca gcagcagtgc cgagacggcc acccggttgt ggtcaacatc
     1501 ctcattggga ggacggactt ccgcgatggc tccattgctg tatagggcct tgtgggtcag
     1561 gacgcttggc tgccttcctg cccctggact gtgtccagct ggtttggagt ctcatcattg
     1621 cttgccctgg gcctatccca tgaggcccag tgactccaca acctccctga ggaggggatg
     1681 gaggggtcaa atctcagaga ggctctcttg tgagctcttg gaccctcctc ttcacggacc
     1741 caactgtgcc gtttgtcccc tctcttatgg agactgaata aaccacctct ggcccatgta
     1801 ggcccaagta ctcagtctca gaacctgtgc atatttattt attatccaca aagatggagg
     1861 tgcggaaaag aagagggaga caggagggaa ccgtactctc ccactggaac tctggggccc
     1921 actgaggcac cattattggg gatttcaggg tggctgggca ctgcaaactg ctccctcctc
     1981 tgtggtccct gaaagagccc cacacgcctg cttcagacgt gtccacgcac accagtcctc
     2041 acagacacac acacacatgc atggaggcaa taaatatgtt ccgtaccaga ctgcccccag
     2101 cctgacgctt cagggggccc cctccaaaag ggaggggttt aagtgctcaa tttttttcgg
     2161 ggggggggca aggggggggc aaggagatgg ggattggaaa ggccagactc tgttatctcc
     2221 atttgctgac tagaggccaa tcgtggggtc ttccccggaa aagggtaggg gaaaccagct
     2281 gtttctctat aagccccagg ctacccctgt gtccatgctg gaacccagga taggggctgc
     2341 cccccacaaa cagtgcctca tgctagtctt tgtgtcgtcc aagggaggga gaaacagtgc
     2401 agtgctcaaa gctaatactg atttggttgg cagcagcaag agggccgact agaggtgact
     2461 caccacacca gggtgatgct cacagtaact ggtgttgccc cggccaaggg gaggcaagag
     2521 aggccagcct agcaagcgcg agaactacct ttttggcaac aaaaggggaa gcccttggtg
     2581 ctcaggaaag atggtcacaa ccgaggctgc ctatcagaag gtggtggctg gtgtggccaa
     2641 tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat
     2701 catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat
     2761 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     2821 aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc
     2881 catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca
     2941 ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt
     3001 gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt
     3061 cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga gatccctcga
     3121 cctgcagccc aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     3181 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3241 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3301 acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3361 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3421 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     3481 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct
     3541 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3601 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatccgc
     3661 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3721 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3781 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3841 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3901 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3961 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4021 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4081 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4141 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4201 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4261 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4321 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4381 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4441 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4501 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4561 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4621 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4681 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4741 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4801 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4861 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4921 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4981 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5041 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5101 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5161 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5221 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5281 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5341 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5401 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5461 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5521 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5581 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5641 cacctgggtc gaca
//
