LOCUS       sequence-205157-        3846 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  T7 promotor drives in vitro mRNA transcription of a mEGFP-tagged 
            membrane reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3846)
  AUTHORS   Clift D, McEwan WA, Labzin LI, Konieczny V, Mogessie B, James LC, 
            Schuh M
  TITLE     A Method for the Acute and Rapid Degradation of Endogenous Proteins.
  JOURNAL   Cell. 2017 Nov 13. pii: S0092-8674(17)31255-2. doi: 
            10.1016/j.cell.2017.10.033.
  PUBMED    29153837
REFERENCE   2  (bases 1 to 3846)
  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"; journalName: "Cell. 2017 
            Nov 13. pii: S0092-8674(17)31255-2. doi: 
            10.1016/j.cell.2017.10.033."
COMMENT     Sequence Label: pGEMHE-membrane-mEGFP
FEATURES             Location/Qualifiers
     source          1..3846
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(53..71)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(54..71)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(89..105)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(89..105)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(102..124)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    113..129
                     /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(137..143,144..161,162..167))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    182..203
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(320..337)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(491..1079)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(571..590)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1250..2041,2042..2110))
                     /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     1873..1892
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2111..2215)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2283..2301
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2339..2361)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2461..2480
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2674..2696
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2688..2705
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2689..2705
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2712..2731
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2712..2730
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           2762..2804
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     regulatory      2882..2891
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2888..2890,2891..2893,2894..3604)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2933..2954)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3194..3213)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     3'UTR           3713..3838
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
ORIGIN
        1 cccctgcagg cgcgccgccg gcgatttaaa ttaattaatc tagcttgagt attctatagt
       61 gtcacctaaa tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
      121 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
      181 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
      241 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
      301 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
      361 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
      421 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
      481 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
      541 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
      601 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
      661 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
      721 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
      781 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
      841 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
      901 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
      961 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1021 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1081 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     1141 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     1201 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     1261 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     1321 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     1381 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     1441 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     1501 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     1561 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     1621 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     1681 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     1741 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     1801 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     1861 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     1921 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     1981 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     2041 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     2101 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     2161 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     2221 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     2281 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     2341 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     2401 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     2461 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     2521 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
     2581 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     2641 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     2701 ccagtgaatt gtaatacgac tcactatagg gcgaattaat tcgagctcgg tacccagctt
     2761 gcttgttctt tttgcagaag ctcagaataa acgctcaact ttggcagatc aattccccgg
     2821 tgctagcatg ggctgcaccg tgagcgccga ggacaaggcg gcggccgagc gcttaccggt
     2881 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     2941 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     3001 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     3061 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     3121 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     3181 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     3241 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     3301 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     3361 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     3421 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     3481 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     3541 catggtcctt aaggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     3601 caagtactca gatctcgagc tcaagcttcg aattctgcag tcgacggccc gggatcctgt
     3661 acaagtaaag cggccgctct agataaatga tagcttgatc tggttaccac taaaccagcc
     3721 tcaagaacac ccgaatggag tctctaagct acataatacc aacttacact ttacaaaatg
     3781 ttgtccccca aaatgtagcc attcgtatct gctcctaata aaaagaaagt ttcttcacat
     3841 tctaaa
//
