LOCUS       sequence-273672-        6407 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 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
FEATURES             Location/Qualifiers
     source          1..6407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     125..145
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    197..277
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      279..288
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(285..287,288..290,291..1004)
                     /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(285..287,288..290,291..1001)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(330..351)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(591..610)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     938..959
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3019..3038)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3084..3139
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3138..3157)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3500..3516)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3500..3516)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3513..3535)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3524..3540
                     /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(3548..3554,3555..3572,3573..3578))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3593..3614
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3673..3868
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3719..3854
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3874..4008
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3911..3930)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3965..3984
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4076..4093)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4247..4835)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4327..4346)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5006..5797,5798..5866))
                     /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     5629..5648
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5867..5971)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6002..6377
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6378..6407,1..174)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
       61 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      121 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta
      181 gtgaaccgtc agatccgcta gcgctaccgg actcagatct cgagctcaag cttcgaattc
      241 tgcagtcgac ggtaccgcgg gcccgggatc caccggtcgc caccatggtg agcaagggcg
      301 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      361 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      421 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
      481 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
      541 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
      601 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
      661 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
      721 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
      781 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
      841 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
      901 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
      961 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaatcaggc ctcacccctc
     1021 ccctgggact ttcccaccca ggaaaggagg accagtcagg atgaggcccc gccttttccc
     1081 ccaccctccc atgagactgc cctgcccagg tatcctgggg gaagaggaga tgtgatcagg
     1141 ccccacccct gtaatcaggc aaggaggagg agtcagatga ggccctgcac cttccccaaa
     1201 ggaaccgccc agtgcaggta tttcagaagg agaaggctgg agaaggacat gagatcaggg
     1261 cctgccccct ggggatcaca gcctcacccc tgcccctgtg ggactcatcc ttgcccaggt
     1321 gagggaagga gacaggatga ggtctcgacc ctgtccccta gggactgtcc tagccaggtc
     1381 tcctgggaaa gggagatgtc aggatgttgc tccatccttt gtcttggaac caccagtcta
     1441 gtccgtcctg gcacagaaga ggagtcaagt aatggaggtc ccagccctgg gggtttaagc
     1501 tctgcccctt ccccatgaac cctgccctgc tctgcccagg caaggaacag aagtgaggat
     1561 gagacccagc cccttcccct gggaactctc ctggccttct aggaatggag gagccaggcc
     1621 ccaccccttc cctataggaa cagcccagca caggtatttc aggtgtgaaa gaatcagtag
     1681 gaccaggcca ccgctagtgc ttgtggagat cacagcccca cccttgtccc tcagcaacat
     1741 cccatctaag cattccacac tgcagggagg agtggtactt aagctcccct gccttaacct
     1801 gggaccaacc tgacctaacc taggagggct ctgagccaac cttgctcttg gggaagggga
     1861 cagattatga aatttcatgg atgaattttc cagacctata tctggagtga gaggccccca
     1921 cccttgggca gagtcctgcc ttcttccttg aggggcagtt tgggaaggtg atgggtatta
     1981 gtgggggact gagttcaggt taccagaacc agtacctcag tattcttttt caacatgtag
     2041 ggcaagagga tgaaggaagg ggctatcctg ggacctcccc agcccaggaa aaactggaag
     2101 ccttccccca gcaaggcaga agcttggagg agggttgtaa aagcatattg taccccctca
     2161 tttgtttatc tgattttttt attgctccgc atactgagaa tctaggccac cccaacctct
     2221 gttccccacc cagttcttca tttggaggaa tcaccccatt tcagagttat caagagacac
     2281 tcccccctcc attcccaccc ctcataccta cacccaaggt tgtcagcttt ggattgctgg
     2341 ggccaggccc catggagggt atactgaggg gtctataggt ttgtgattaa aataataaaa
     2401 gctaggcgtg tttgatgcgc ttttaacttt ggcaaagagt ctcctgtcct tgttttacac
     2461 tgtgcagtga acatagccca tgcaggccaa gagggataaa agtccttggg agaaacagat
     2521 tccagaagga tggataaaag aatttagaaa ggaagagggc atggaggaag acaatatgca
     2581 aacaaaagac agtattttta aagaccaatc accaggcaca attttaactg ctgcaagtgt
     2641 ttcatctaat ttatttctct aaagggatgt tagtcagaac atgctgcttc ttacttccat
     2701 ttggtctttg aagaagctga ggaatagaat tgaactgttc tgaagatatg caagcctgta
     2761 aatgggtgag gcggtaaaac agtaaagtca ggtgagtgag ggctcgattg aaaaatgcca
     2821 ctgagggcag agggggaaag aagcttttct tgcaaatcca cttttgaaac tgataaagaa
     2881 gtctatgagg atatttcctg gcagacaaca gaggtattta agatggtgga aaggcggccg
     2941 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     3001 tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc
     3061 ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     3121 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca
     3181 gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa
     3241 aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt
     3301 ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt
     3361 ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc
     3421 ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc
     3481 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3541 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     3601 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3661 gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3721 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3781 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3841 cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt
     3901 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3961 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat
     4021 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4081 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4141 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4201 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4261 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4321 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4381 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4441 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4501 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4561 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4621 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4681 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4741 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4801 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4861 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4921 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4981 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5041 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5101 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5161 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5221 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5281 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5341 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5401 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5461 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5521 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5581 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5641 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5701 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5761 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5821 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5881 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5941 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt
     6001 cgacattgat tattgactag taatagtaat caattacggg gtcattagtt catagcccat
     6061 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     6121 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     6181 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     6241 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     6301 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     6361 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatggg
//
