LOCUS       sequence-274554-        6407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human TFE3 poly(A) site reporter, stem loop mutant.
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
FEATURES             Location/Qualifiers
     source          1..6407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        327..530
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     481..501
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    553..633
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      635..644
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(641..643,644..646,647..1360)
                     /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(641..643,644..646,647..1357)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(686..707)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(947..966)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1294..1315
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3375..3394)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3440..3495
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3494..3513)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3856..3872)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3856..3872)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3869..3891)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3880..3896
                     /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(3904..3910,3911..3928,3929..3934))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3949..3970
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4029..4224
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4075..4210
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4230..4364
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4267..4286)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4321..4340
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4432..4449)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4603..5191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4683..4702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5362..6153,6154..6222))
                     /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     5985..6004
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6223..6327)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(6358..6407,1..326)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
       61 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      121 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      181 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      241 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      301 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      361 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      421 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      481 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
      541 accgtcagat ccgctagcgc taccggactc agatctcgag ctcaagcttc gaattctgca
      601 gtcgacggta ccgcgggccc gggatccacc ggtcgccacc atggtgagca agggcgagga
      661 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      721 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
      781 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
      841 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
      901 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
      961 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1021 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1081 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     1141 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     1201 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     1261 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1321 cgccgggatc actctcggca tggacgagct gtacaagtaa tcaggcctca cccctcccct
     1381 gggactttcc cacccaggaa aggaggacca gtcaggatga ggccccgcct tttcccccac
     1441 cctcccatga gactgccctg cccaggtatc ctgggggaag aggagatgtg atcaggcccc
     1501 acccctgtaa tcaggcaagg aggaggagtc agatgaggcc ctgcaccttc cccaaaggaa
     1561 ccgcccagtg caggtatttc agaaggagaa ggctggagaa ggacatgaga tcagggcctg
     1621 ccccctgggg atcacagcct cacccctgcc cctgtgggac tcatccttgc ccaggtgagg
     1681 gaaggagaca ggatgaggtc tcgaccctgt cccctaggga ctgtcctagc caggtctcct
     1741 gggaaaggga gatgtcagga tgttgctcca tcctttgtct tggaaccacc agtctagtcc
     1801 gtcctggcac agaagaggag tcaagtaatg gaggtcccag ccctgggggt ttaagctctg
     1861 ccccttcccc atgaaccctg ccctgctctg cccaggcaag gaacagaagt gaggatgaga
     1921 cccagcccct tcccctggga actctcctgg ccttctagga atggaggagc caggccccac
     1981 cccttcccta taggaacagc ccagcacagg tatttcaggt gtgaaagaat cagtaggacc
     2041 aggccaccgc tagtgcttgt ggagatcaca gccccaccct tgtccctcag caacatccca
     2101 tctaagcatt ccacactgca gggaggagtg gtacttaagc tcccctgcct taacctggga
     2161 ccaacctgac ctaacctagg agggctctga gccaaccttg ctcttgggga aggggacaga
     2221 ttatgaaatt tcatggatga attttccaga cctatatctg gagtgagagg cccccaccct
     2281 tgggcagagt cctgccttct tccttgaggg gcagtttggg aaggtgatgg gtattagtgg
     2341 gggactgagt tcaggttacc agaaccagta cctcagtatt ctttttcaac atgtagggca
     2401 agaggatgaa ggaaggggct atcctgggac ctccccagcc caggaaaaac tggaagcctt
     2461 cccccagcaa ggcagaagct tggaggaggg ttgtaaaagc atattgtacc ccctcatttg
     2521 tttatctgat ttttttattg ctccgcatac tgagaatcta ggccacccca acctctgttc
     2581 cccacccagt tcttcatttg gaggaatcac cccatttcag agttatcaag agacactccc
     2641 ccctccattc ccacccctca tacctacacc caaggttgtc agctttggat tgctggggcc
     2701 aggccccatg gagggtatac tgaggggtct ataggtttgt gattaaaata ataaaagcta
     2761 ggcctgtttg atgcgctttt aactttggca aagagtctcc tgtccttgtt ttacactgtg
     2821 cagtgaacat agcccatgca ggccaagagg gataaaagtc cttgggagaa acagattcca
     2881 gaaggatgga taaaagaatt tagaaaggaa gagggcatgg aggaagacaa tatgcaaaca
     2941 aaagacagta tttttaaaga ccaatcacca ggcacaattt taactgctgc aagtgtttca
     3001 tctaatttat ttctctaaag ggatgttagt cagaacatgc tgcttcttac ttccatttgg
     3061 tctttgaaga agctgaggaa tagaattgaa ctgttctgaa gatatgcaag cctgtaaatg
     3121 ggtgaggcgg taaaacagta aagtcaggtg agtgagggct cgattgaaaa atgccactga
     3181 gggcagaggg ggaaagaagc ttttcttgca aatccacttt tgaaactgat aaagaagtct
     3241 atgaggatat ttcctggcag acaacagagg tatttaagat ggtggaaagg cggccgcact
     3301 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac
     3361 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     3421 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     3481 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     3541 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     3601 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     3661 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     3721 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     3781 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag
     3841 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3901 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3961 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4021 gcggatccgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4081 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4141 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4201 tttggaggcc taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca
     4261 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4321 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat
     4381 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4441 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4501 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4561 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4621 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4681 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4741 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4801 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4861 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4921 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4981 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5041 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5101 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5161 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5221 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5281 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5341 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5401 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5461 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5521 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5581 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5641 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5701 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5761 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5821 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5881 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5941 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6001 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6061 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6121 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6181 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6241 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6301 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcgac
     6361 attgattatt gactagtaat agtaatcaat tacggggtca ttagttc
//
