LOCUS       sequence-274587-        5653 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human ILVBL poly(A) site reporter, stem loop mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5653)
  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 5653)
  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-G10C
FEATURES             Location/Qualifiers
     source          1..5653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        350..553
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     504..524
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    576..656
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      658..667
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(664..666,667..669,670..1383)
                     /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(709..730)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(970..989)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1317..1338
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2644..2663)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2709..2764
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2763..2782)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3125..3141)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3125..3141)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3138..3160)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3149..3165
                     /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(3173..3179,3180..3197,3198..3203))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3218..3239
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3298..3493
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3344..3479
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3499..3633
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3536..3555)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3590..3609
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3701..3718)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3872..4460)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3952..3971)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4631..5422,5423..5491))
                     /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     5254..5273
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5492..5596)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(5627..5653,1..349)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
       61 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      121 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      181 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      241 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      301 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
      361 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      421 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      481 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      541 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga ctcagatctc
      601 gagctcaagc ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc accggtcgcc
      661 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      721 gacggcgacg taaacgacca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      781 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      841 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
      901 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      961 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1021 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1081 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     1141 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     1201 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1261 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     1321 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1381 taaagcggcc gcactcctca ggtgcgatca ggtggtcaag gtgctgcacg atgcccagca
     1441 gcagtgccga gacggccacc cggttgtggt caacatcctc attgggagga cggacttccg
     1501 cgatggctcc attgctgtat agggccttgt gggtcaggac gcttggctgc cttcctgccc
     1561 ctggactgtg tccagctggt ttggagtctc atcattgctt gccctgggcc tatcccatga
     1621 ggcccagtga ctccacaacc tccctgagga ggggatggag gggtcaaatc tcagagaggc
     1681 tctcttgtga gctcttggac cctcctcttc acggacccaa ctgtgccgtt tgtcccctct
     1741 cttatggaga ctgaataaac cacctctgcc ccatgtaggc ccaagtactc agtctcagaa
     1801 cctgtgcata tttatttatt atccacaaag atggaggtgc ggaaaagaag agggagacag
     1861 gagggaaccg tactctccca ctggaactct ggggcccact gaggcaccat tattggggat
     1921 ttcagggtgg ctgggcactg caaactgctc cctcctctgt ggtccctgaa agagccccac
     1981 acgcctgctt cagacgtgtc cacgcacacc agtcctcaca gacacacaca cacatgcatg
     2041 gaggcaataa atatgttccg taccagactg cccccagcct gacgcttcag ggggccccct
     2101 ccaaaaggga ggggtttaag tgctcaattt ttttcggggg gggggcaagg gggggcaagg
     2161 agatggggat tggaaaggcc agactctgtt atctccattt gctgactaga ggccaatcgt
     2221 ggggtcttcc ccggaaaagg gtaggggaaa ccagctgttt ctctataagc cccaggctac
     2281 ccctgtgtcc atgctggaac ccaggatagg ggctgccccc cacaaacagt gcctcatgct
     2341 agtctttgtg tcgtccaagg gagggagaaa cagtgcagtg ctcaaagcta atactgattt
     2401 ggttggcagc agcaagaggg ccgactagag gtgactcacc acaccagggt gatgctcaca
     2461 gtaactggtg ttgccccggc caaggggagg caagagaggc cagcctagca agcgcgagaa
     2521 ctaccttttt ggcaacaaaa ggggaagccc ttggtgctca ggaaagatgg tcacaaccga
     2581 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg
     2641 agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac
     2701 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     2761 ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt
     2821 ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag
     2881 aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt
     2941 gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct
     3001 aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt
     3061 catagctgtc cctcttctct tatggagatc cctcgacctg cagcccaagc ttggcgtaat
     3121 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     3181 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     3241 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatccgca
     3301 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3361 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3421 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3481 aggcttttgc aaaaagctaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3541 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3601 aaactcatca atgtatctta tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg
     3661 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3721 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3781 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3841 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3901 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3961 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4021 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4081 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4141 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4201 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4261 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4321 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4381 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4441 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4501 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4561 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4621 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4681 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4741 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4801 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4861 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4921 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4981 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5041 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5101 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5161 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5221 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5281 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5341 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5401 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5461 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5521 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5581 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca ttgattattg
     5641 actagtaata gta
//
