LOCUS       sequence-274555-        6407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human TFE3 poly(A) site reporter, stem loop mutant rescue.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6407)
  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 6407)
  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-TFE3pA-G9C-C19G
FEATURES             Location/Qualifiers
     source          1..6407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        339..542
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     493..513
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    565..645
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      647..656
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(653..655,656..658,659..1372)
                     /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(653..655,656..658,659..1369)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(698..719)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(959..978)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1306..1327
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3387..3406)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3452..3507
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3506..3525)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3868..3884)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3868..3884)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3881..3903)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3892..3908
                     /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(3916..3922,3923..3940,3941..3946))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3961..3982
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4041..4236
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4087..4222
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4242..4376
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4279..4298)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4333..4352
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4444..4461)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4615..5203)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4695..4714)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5374..6165,6166..6234))
                     /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     5997..6016
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6235..6339)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(6370..6407,1..338)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
       61 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      121 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      181 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      241 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      301 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      361 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      421 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      481 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      541 ctggtttagt gaaccgtcag atccgctagc gctaccggac tcagatctcg agctcaagct
      601 tcgaattctg cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca ccatggtgag
      661 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
      721 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
      781 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
      841 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
      901 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
      961 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1021 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1081 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     1141 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     1201 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1261 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1321 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aatcaggcct
     1381 cacccctccc ctgggacttt cccacccagg aaaggaggac cagtcaggat gaggccccgc
     1441 cttttccccc accctcccat gagactgccc tgcccaggta tcctggggga agaggagatg
     1501 tgatcaggcc ccacccctgt aatcaggcaa ggaggaggag tcagatgagg ccctgcacct
     1561 tccccaaagg aaccgcccag tgcaggtatt tcagaaggag aaggctggag aaggacatga
     1621 gatcagggcc tgccccctgg ggatcacagc ctcacccctg cccctgtggg actcatcctt
     1681 gcccaggtga gggaaggaga caggatgagg tctcgaccct gtcccctagg gactgtccta
     1741 gccaggtctc ctgggaaagg gagatgtcag gatgttgctc catcctttgt cttggaacca
     1801 ccagtctagt ccgtcctggc acagaagagg agtcaagtaa tggaggtccc agccctgggg
     1861 gtttaagctc tgccccttcc ccatgaaccc tgccctgctc tgcccaggca aggaacagaa
     1921 gtgaggatga gacccagccc cttcccctgg gaactctcct ggccttctag gaatggagga
     1981 gccaggcccc accccttccc tataggaaca gcccagcaca ggtatttcag gtgtgaaaga
     2041 atcagtagga ccaggccacc gctagtgctt gtggagatca cagccccacc cttgtccctc
     2101 agcaacatcc catctaagca ttccacactg cagggaggag tggtacttaa gctcccctgc
     2161 cttaacctgg gaccaacctg acctaaccta ggagggctct gagccaacct tgctcttggg
     2221 gaaggggaca gattatgaaa tttcatggat gaattttcca gacctatatc tggagtgaga
     2281 ggcccccacc cttgggcaga gtcctgcctt cttccttgag gggcagtttg ggaaggtgat
     2341 gggtattagt gggggactga gttcaggtta ccagaaccag tacctcagta ttctttttca
     2401 acatgtaggg caagaggatg aaggaagggg ctatcctggg acctccccag cccaggaaaa
     2461 actggaagcc ttcccccagc aaggcagaag cttggaggag ggttgtaaaa gcatattgta
     2521 ccccctcatt tgtttatctg atttttttat tgctccgcat actgagaatc taggccaccc
     2581 caacctctgt tccccaccca gttcttcatt tggaggaatc accccatttc agagttatca
     2641 agagacactc ccccctccat tcccacccct catacctaca cccaaggttg tcagctttgg
     2701 attgctgggg ccaggcccca tggagggtat actgaggggt ctataggttt gtgattaaaa
     2761 taataaaagc taggcctgtt tgatgggctt ttaactttgg caaagagtct cctgtccttg
     2821 ttttacactg tgcagtgaac atagcccatg caggccaaga gggataaaag tccttgggag
     2881 aaacagattc cagaaggatg gataaaagaa tttagaaagg aagagggcat ggaggaagac
     2941 aatatgcaaa caaaagacag tatttttaaa gaccaatcac caggcacaat tttaactgct
     3001 gcaagtgttt catctaattt atttctctaa agggatgtta gtcagaacat gctgcttctt
     3061 acttccattt ggtctttgaa gaagctgagg aatagaattg aactgttctg aagatatgca
     3121 agcctgtaaa tgggtgaggc ggtaaaacag taaagtcagg tgagtgaggg ctcgattgaa
     3181 aaatgccact gagggcagag ggggaaagaa gcttttcttg caaatccact tttgaaactg
     3241 ataaagaagt ctatgaggat atttcctggc agacaacaga ggtatttaag atggtggaaa
     3301 ggcggccgca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
     3361 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
     3421 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
     3481 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
     3541 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
     3601 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
     3661 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
     3721 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
     3781 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac
     3841 ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     3901 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3961 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4021 cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4081 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4141 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     4201 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt
     4261 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4321 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct
     4381 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4441 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4501 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4561 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4621 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4681 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4741 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4801 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4861 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4921 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4981 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5041 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5101 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5161 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5221 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5281 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5341 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5401 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5461 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5521 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5581 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5641 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5701 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5761 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5821 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5881 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5941 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6001 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6061 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6121 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6181 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6241 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6301 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6361 acctgggtcg acattgatta ttgactagta atagtaatca attacgg
//
