LOCUS       sequence-227877-        5650 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 5650)
  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 5650)
  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..5650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        376..579
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     530..550
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    602..682
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      684..693
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(690..692,693..695,696..1409)
                     /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(690..692,693..695,696..1406)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(735..756)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(996..1015)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1343..1364
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2672..2691)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2737..2792
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2791..2810)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3153..3169)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3153..3169)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3166..3188)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3177..3193
                     /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(3201..3207,3208..3225,3226..3231))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3246..3267
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3326..3521
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3372..3507
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3527..3661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3564..3583)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3618..3637
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3729..3746)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3900..4488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3980..3999)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4659..5450,5451..5519))
                     /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     5282..5301
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5520..5624)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgata atagtaatca attacggggt cattagttca tagcccatat
       61 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      121 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      181 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      241 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      301 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      361 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      421 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      481 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      541 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      601 cgctagcgct accggactca gatctcgagc tcaagcttcg aattctgcag tcgacggtac
      661 cgcgggcccg ggatccaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
      721 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
      781 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
      841 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
      901 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
      961 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1021 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1081 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1141 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     1201 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1261 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     1321 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1381 ctctcggcat ggacgagctg tacaagtaaa gcggccgcac tcctcaggtg cgatcaggtg
     1441 gtcaaggtgc tgcacgatgc ccagcagcag tgccgagacg gccacccggt tgtggtcaac
     1501 atcctcattg ggaggacgga cttccgcgat ggctccattg ctgtataggg ccttgtgggt
     1561 caggacgctt ggctgccttc ctgcccctgg actgtgtcca gctggtttgg agtctcatca
     1621 ttgcttgccc tgggcctatc ccatgaggcc cagtgactcc acaacctccc tgaggagggg
     1681 atggaggggt caaatctcag agaggctctc ttgtgagctc ttggaccctc ctcttcacgg
     1741 acccaactgt gccgtttgtc ccctctctta tggagactga ataaaccacc tctggcccat
     1801 gtaggcccaa gtactcagtc tcagaacctg tgcatattta tttattatcc acaaagatgg
     1861 aggtgcggaa aagaagaggg agacaggagg gaaccgtact ctcccactgg aactctgggg
     1921 cccactgagg caccattatt ggggatttca gggtggctgg gcactgcaaa ctgctccctc
     1981 ctctgtggtc cctgaaagag ccccacacgc ctgcttcaga cgtgtccacg cacaccagtc
     2041 ctcacagaca cacacacaca tgcatggagg caataaatat gttccgtacc agactgcccc
     2101 cagcctgacg cttcaggggg ccccctccaa aagggagggg tttaagtgct caattttttt
     2161 cggggggggg ggcaaggggg gggcaaggag atggggattg gaaaggccag actctgttat
     2221 ctccatttgc tgactagagg ccaatcgtgg ggacttcccc ggaaaagggt aggggaaacc
     2281 agctgtttct ctataagccc caggctaccc ctgtgtccat gctggaaccc aggatagggg
     2341 ctgcccccca caaacagtgc ctcatgctag tctttgtgtc gtccaaggga gggagaaaca
     2401 gtgcagtgct caaagctaat actgatttgg ttggcagcag caagagggcc gactagaggt
     2461 gactcaccac accagggtga tgctcacagt aactggtgtt gccccggcca aggggaggca
     2521 agagaggcca gcctagcaag cgcgagaact acctttttgg caacaaaagg ggaagccctt
     2581 ggtgctcagg aaagatggtc acaaccgagg ctgcctatca gaaggtggtg gctggtgtgg
     2641 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     2701 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     2761 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     2821 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     2881 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     2941 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     3001 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     3061 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc
     3121 tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     3181 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3241 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3301 ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc
     3361 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3421 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3481 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc
     3541 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3601 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     3661 ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3721 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3781 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3841 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3901 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3961 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4021 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4081 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4141 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4201 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4261 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4321 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4381 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4441 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4501 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4561 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4621 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4681 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4741 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4801 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4861 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4921 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4981 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5041 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5101 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5161 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5221 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5281 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5341 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5401 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5461 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5521 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5581 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5641 gtgccacctg
//
