LOCUS       sequence-227864-        6400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human TFE3 poly(A) site reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6400)
  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 6400)
  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
FEATURES             Location/Qualifiers
     source          1..6400
                     /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(3424..3443)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3489..3544
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3543..3562)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3903..3919)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3903..3919)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3916..3938)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3927..3943
                     /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(3951..3957,3958..3975,3976..3981))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3996..4017
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4076..4271
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4122..4257
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4277..4411
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4314..4333)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4368..4387
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4479..4496)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4650..5238)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4730..4749)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5409..6200,6201..6269))
                     /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     6032..6051
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6270..6374)
                     /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 tacaagtaat caggcctcac ccctcccctg ggactttccc
     1441 acccaggaaa ggaggaccag tcaggatgag gccccgcctt ttcccccacc ctcccatgag
     1501 actgccctgc ccaggtatcc tgggggaaga ggagatgtga tcaggcccca cccctgtaat
     1561 caggcaagga ggaggagtca gatgaggccc tgcaccttcc ccaaaggaac cgcccagtgc
     1621 aggtatttca gaaggagaag gctggagaag gacatgagat cagggcctgc cccctgggga
     1681 tcacagcctc acccctgccc ctgtgggact catccttgcc caggtgaggg aaggagacag
     1741 gatgaggtct cgaccctgtc ccctagggac tgtcctagcc aggtctcctg ggaaagggag
     1801 atgtcaggat gttgctccat cctttgtctt ggaaccacca gtctagtccg tcctggcaca
     1861 gaagaggagt caagtaatgg aggtcccagc cctgggggtt taagctctgc cccttcccca
     1921 tgaaccctgc cctgctctgc ccaggcaagg aacagaagtg aggatgagac ccagcccctt
     1981 cccctgggaa ctctcctggc cttctaggaa tggaggagcc aggccccacc ccttccctat
     2041 aggaacagcc cagcacaggt atttcaggtg tgaaagaatc agtaggacca ggccaccgct
     2101 agtgcttgtg gagatcacag ccccaccctt gtccctcagc aacatcccat ctaagcattc
     2161 cacactgcag ggaggagtgg tacttaagct cccctgcctt aacctgggac caacctgacc
     2221 taacctagga gggctctgag ccaaccttgc tcttggggaa ggggacagat tatgaaattt
     2281 catggatgaa ttttccagac ctatatctgg agtgagaggc ccccaccctt gggcagagtc
     2341 ctgccttctt ccttgagggg cagtttggga aggtgatggg tattagtggg ggactgagtt
     2401 caggttacca gaaccagtac ctcagtattc tttttcaaca tgtagggcaa gaggatgaag
     2461 gaaggggcta tcctgggacc tccccagccc aggaaaaact ggaagccttc ccccagcaag
     2521 gcagaagctt ggaggagggt tgtaaaagca tattgtaccc cctcatttgt ttatctgatt
     2581 tttttattgc tccgcatact gagaatctag gccaccccaa cctctgttcc ccacccagtt
     2641 cttcatttgg aggaatcacc ccatttcaga gttatcaaga gacactcccc cctccattcc
     2701 cacccctcat acctacaccc aaggttgtca gctttggatt gctggggcca ggccccatgg
     2761 agggtatact gaggggtcta taggtttgtg attaaaataa taaaagctag gcgtgtttga
     2821 tgcgctttta actttggcaa agagtctcct gtccttgttt tacactgtgc agtgaacata
     2881 gcccatgcag gccaagaggg ataaaagtcc ttgggagaaa cagattccag aaggatggat
     2941 aaaagaattt agaaaggaag agggcatgga ggaagacaat atgcaaacaa aagacagtat
     3001 ttttaaagac caatcaccag gcacaatttt aactgctgca agtgtttcat ctaatttatt
     3061 tctctaaagg gatgttagtc agaacatgct gcttcttact tccatttggt ctttgaagaa
     3121 gctgaggaat agaattgaac tgttctgaag atatgcaagc ctgtaaatgg gtgaggcggt
     3181 aaaacagtaa agtcaggtga gtgagggctc gattgaaaaa tgccactgag ggcagagggg
     3241 gaaagaagct tttcttgcaa atccactttt gaaactgata aagaagtcta tgaggatatt
     3301 tcctggcaga caacagaggt atttaagatg gtggaaaggc ggccgcactc ctcaggtgca
     3361 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg
     3421 agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac
     3481 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     3541 ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt
     3601 ttagagtttg gcaacatatg ccatatgctg gctgccatga acaaaggtgg ctataaagag
     3661 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga
     3721 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa
     3781 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca
     3841 tagctgtccc tcttctctta tgaagatccc tcgacctgca gcccaagctt ggcgtaatca
     3901 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     3961 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4021 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc
     4081 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4141 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4201 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4261 gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4321 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4381 actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg
     4441 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4501 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4561 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4621 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4681 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4741 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4801 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     4861 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4921 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4981 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5041 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5101 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5161 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5221 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5281 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5341 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5401 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5461 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5521 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5581 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5641 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5701 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5761 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5821 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5881 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5941 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6001 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6061 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6121 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6181 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6241 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6301 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6361 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
