LOCUS       sequence-274559-        6388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  human TFE3 poly(A) site reporter, with stem loop deleted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6388)
  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 6388)
  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-del
FEATURES             Location/Qualifiers
     source          1..6388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     220..241
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2282..2301)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2347..2402
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2401..2420)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2763..2779)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2763..2779)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2776..2798)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2787..2803
                     /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(2811..2817,2818..2835,2836..2841))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2856..2877
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2936..3131
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2982..3117
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3137..3271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3174..3193)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3228..3247
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3339..3356)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3510..4098)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3590..3609)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4269..5060,5061..5129))
                     /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     4892..4911
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5130..5234)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5265..5640
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5641..5844
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5795..5815
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    5867..5947
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      5949..5958
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5955..5957,5958..5960,5961..6388,1..286)
                     /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(5955..5957,5958..5960,5961..6388,1..283)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6000..6021)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6261..6280)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
       61 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
      121 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
      181 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
      241 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaatcag gcctcacccc
      301 tcccctggga ctttcccacc caggaaagga ggaccagtca ggatgaggcc ccgccttttc
      361 ccccaccctc ccatgagact gccctgccca ggtatcctgg gggaagagga gatgtgatca
      421 ggccccaccc ctgtaatcag gcaaggagga ggagtcagat gaggccctgc accttcccca
      481 aaggaaccgc ccagtgcagg tatttcagaa ggagaaggct ggagaaggac atgagatcag
      541 ggcctgcccc ctggggatca cagcctcacc cctgcccctg tgggactcat ccttgcccag
      601 gtgagggaag gagacaggat gaggtctcga ccctgtcccc tagggactgt cctagccagg
      661 tctcctggga aagggagatg tcaggatgtt gctccatcct ttgtcttgga accaccagtc
      721 tagtccgtcc tggcacagaa gaggagtcaa gtaatggagg tcccagccct gggggtttaa
      781 gctctgcccc ttccccatga accctgccct gctctgccca ggcaaggaac agaagtgagg
      841 atgagaccca gccccttccc ctgggaactc tcctggcctt ctaggaatgg aggagccagg
      901 ccccacccct tccctatagg aacagcccag cacaggtatt tcaggtgtga aagaatcagt
      961 aggaccaggc caccgctagt gcttgtggag atcacagccc cacccttgtc cctcagcaac
     1021 atcccatcta agcattccac actgcaggga ggagtggtac ttaagctccc ctgccttaac
     1081 ctgggaccaa cctgacctaa cctaggaggg ctctgagcca accttgctct tggggaaggg
     1141 gacagattat gaaatttcat ggatgaattt tccagaccta tatctggagt gagaggcccc
     1201 cacccttggg cagagtcctg ccttcttcct tgaggggcag tttgggaagg tgatgggtat
     1261 tagtggggga ctgagttcag gttaccagaa ccagtacctc agtattcttt ttcaacatgt
     1321 agggcaagag gatgaaggaa ggggctatcc tgggacctcc ccagcccagg aaaaactgga
     1381 agccttcccc cagcaaggca gaagcttgga ggagggttgt aaaagcatat tgtaccccct
     1441 catttgttta tctgattttt ttattgctcc gcatactgag aatctaggcc accccaacct
     1501 ctgttcccca cccagttctt catttggagg aatcacccca tttcagagtt atcaagagac
     1561 actcccccct ccattcccac ccctcatacc tacacccaag gttgtcagct ttggattgct
     1621 ggggccaggc cccatggagg gtatactgag gggtctatag gtttgtgatt aaaataataa
     1681 aagctttaac tttggcaaag agtctcctgt ccttgtttta cactgtgcag tgaacatagc
     1741 ccatgcaggc caagagggat aaaagtcctt gggagaaaca gattccagaa ggatggataa
     1801 aagaatttag aaaggaagag ggcatggagg aagacaatat gcaaacaaaa gacagtattt
     1861 ttaaagacca atcaccaggc acaattttaa ctgctgcaag tgtttcatct aatttatttc
     1921 tctaaaggga tgttagtcag aacatgctgc ttcttacttc catttggtct ttgaagaagc
     1981 tgaggaatag aattgaactg ttctgaagat atgcaagcct gtaaatgggt gaggcggtaa
     2041 aacagtaaag tcaggtgagt gagggctcga ttgaaaaatg ccactgaggg cagaggggga
     2101 aagaagcttt tcttgcaaat ccacttttga aactgataaa gaagtctatg aggatatttc
     2161 ctggcagaca acagaggtat ttaagatggt ggaaaggcgg ccgcactcct caggtgcagg
     2221 ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag
     2281 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     2341 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     2401 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
     2461 agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag
     2521 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga
     2581 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa
     2641 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca
     2701 tagctgtccc tcttctctta tggagatccc tcgacctgca gcccaagctt ggcgtaatca
     2761 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     2821 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     2881 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc
     2941 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3001 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3061 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3121 gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3181 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3241 actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg
     3301 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3361 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3421 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3481 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3541 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3601 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3661 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3721 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3781 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3841 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3901 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3961 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4021 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4081 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4141 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4201 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4261 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4321 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4381 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4441 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4501 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4561 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4621 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4681 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4741 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4801 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4861 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4921 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4981 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5041 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5101 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5161 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5221 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ggtcgacatt gattattgac
     5281 tagtaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     5341 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     5401 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     5461 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     5521 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     5581 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     5641 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     5701 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     5761 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     5821 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
     5881 actcagatct cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc
     5941 caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     6001 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     6061 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     6121 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     6181 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     6241 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     6301 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     6361 acatcctggg gcacaagctg gagtacaa
//
