LOCUS       sequence-205152-        7206 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  T7 promotor drives in vitro transcription of mEGFP-tagged Histon 
            MAP4 mRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7206)
  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 7206)
  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-mEGFP-Map4
FEATURES             Location/Qualifiers
     source          1..7206
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(52..70)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(53..70)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(88..104)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(88..104)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(101..123)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    112..128
                     /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(136..142,143..160,161..166))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..202
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(319..336)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(490..1078)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(570..589)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1249..2040,2041..2109))
                     /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     1872..1891
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2110..2214)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2282..2300
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2338..2360)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2460..2479
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2673..2695
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2687..2704
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2688..2704
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2711..2730
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2711..2729
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           2761..2803
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     regulatory      2837..2846
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2843..2845,2846..2848,2849..3559)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2843..2845,2846..2848,2849..3559)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2888..2909)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3149..3168)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3496..3517
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     3'UTR           7078..7203
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
ORIGIN
        1 ccctgcaggc gcgccgccgg cgatttaaat taattaatct agcttgagta ttctatagtg
       61 tcacctaaat agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
      121 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
      181 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
      241 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
      301 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
      361 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
      421 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      481 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      541 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
      601 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
      661 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
      721 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
      781 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
      841 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
      901 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
      961 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1021 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1081 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1141 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1201 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1261 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     1321 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1381 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     1441 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     1501 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     1561 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     1621 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     1681 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     1741 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     1801 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     1861 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     1921 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     1981 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     2041 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     2101 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     2161 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     2221 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     2281 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     2341 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     2401 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     2461 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     2521 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     2581 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     2641 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     2701 cagtgaattg taatacgact cactataggg cgaattaatt cgagctcggt acccagcttg
     2761 cttgttcttt ttgcagaagc tcagaataaa cgctcaactt tggcagatca attccccggt
     2821 gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     2881 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2941 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     3001 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     3061 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     3121 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     3181 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3241 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     3301 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     3361 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3421 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     3481 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     3541 catggacgag ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt
     3601 cgacagaatg gccgacctca gtcttgtgga tgcgttgaca gaaccacctc cagaaattga
     3661 gggagaaata aagcgagact tcatggctgc gctggaggca gagccctatg atgacatcgt
     3721 gggagaaact gtggagaaaa ctgagtttat tcctctcctg gatggtgatg agaaaaccgg
     3781 gaactcagag tccaaaaaga aaccctgctt agacactagc caggttgaag gtatcccatc
     3841 ttctaaacca acactcctag ccaatggtga tcatggaatg gaggggaata acactgcagg
     3901 gtctccaact gacttccttg aagagagagt ggactatccg gattatcaga gcagccagaa
     3961 ctggccagaa gatgcaagct tttgtttcca gcctcagcaa gtgttagata ctgaccaggc
     4021 tgagcccttt aacgagcacc gtgatgatgg tttggcagat ctgctctttg tctccagtgg
     4081 acccacgaac gcttctgcat ttacagagcg agacaatcct tcagaagaca gttacggtat
     4141 gcttccctgt gactcatttg cttccacggc tgttgtatct caggagtggt ctgtgggagc
     4201 cccaaactct ccatgttcag agtcctgtgt ctccccagag gttactatag aaaccctaca
     4261 gccagcaaca gagctctcca aggcagcaga agtggaatca gtgaaagagc agctgccagc
     4321 taaagcattg gaaacgatgg cagagcagac cactgatgtg gtgcactctc catccacaga
     4381 cacaacacca ggcccagaca cagaggcagc actggctaaa gacatagaag agatcaccaa
     4441 gccagatgtg atattggcaa atgtcacgca gccatctact gaatcggata tgttcctggc
     4501 ccaggacatg gaactactca caggaacaga ggcagcccac gctaacaata tcatattgcc
     4561 tacagaacca gacgaatctt caaccaagga tgtagcacca cctatggaag aagaaattgt
     4621 cccaggcaat gatacgacat cccccaaaga aacagagaca acacttccaa taaaaatgga
     4681 cttggcacca cctgaggatg tgttacttac caaagaaaca gaactagccc cagccaaggg
     4741 catggtttca ctctcagaaa tagaagaggc tctggcaaag aatgatgagt cctctgcaga
     4801 aatacctgtg gctcaggaga cagtggtctc agaaacagag gtggtcctgg caacagaagt
     4861 ggtactgccc tcagatccca taacaacatt gacaaaggat gtgacactcc ccttagaagc
     4921 agagagaccg ttggtgacgg acatgactcc atctctggaa acagaaatga ccctaggcaa
     4981 agagacagct ccacccacag aaacaaattt gggcatggcc aaagacatgt ctccactccc
     5041 agaatcagaa gtgactctgg gcaaggacgt ggttatactt ccagaaacaa aggtggctga
     5101 gtttaacaat gtgactccac tttcagaaga agaggtaacc tcagtcaagg acatgtctcc
     5161 gtctgcagaa acagaggctc ccctggctaa gaatgctgat ctgcactcag gaacagagct
     5221 gattgtggac aacagcatgg ctccagcctc cgatcttgca ctgcccttgg aaacaaaagt
     5281 agcaacagtt ccaattaaag acaaaggaac tgtacagact gaagaaaaac cacgtgaaga
     5341 ctcccagtta gcatctatgc agcacaaggg acagtcaaca gtacctcctt gcacggcttc
     5401 accagaacca gtcaaagctg cagaacaaat gtctacctta ccaatagatg caccttctcc
     5461 attagagaac ttagagcaga aggaaacgcc tggcagccag ccttctgagc cttgctcagg
     5521 agtatcccgg caagaagaag caaaggctgc tgtaggtgtg actggaaatg acatcactac
     5581 cccgccaaac aaggagccac caccaagccc agaaaagaaa gcaaagcctt tggccaccac
     5641 tcaacctgca aagacttcaa catcgaaagc caaaacacag cccacttctc tccctaagca
     5701 accagctccc accacctctg gtgggttgaa taaaaaaccc atgagcctcg cctcaggctc
     5761 agtgccagct gccccacaca aacgccctgc tgctgccact gctactgcca ggccttccac
     5821 cctacctgcc agagacgtga agccaaagcc aattacagaa gctaaggttg ccgaaaagcg
     5881 gacctctcca tccaagcctt catctgcccc agccctcaaa cctggaccta aaaccacccc
     5941 aaccgtttca aaagccacat ctccctcaac tcttgtttcc actggaccaa gtagtagaag
     6001 tccagctaca actctgccta agaggccaac cagcatcaag actgagggga aacctgctga
     6061 tgtcaaaagg atgactgcta agtctgcctc agctgacttg agtcgctcaa agaccacctc
     6121 tgccagttct gtgaagagaa acaccactcc cactggggca gcacccccag cagggatgac
     6181 ttccactcga gtcaagccca tgtctgcacc tagccgctct tctggggctc tttctgtgga
     6241 caagaagccc acttccacta agcctagctc ctctgctccc agggtgagcc gcctggccac
     6301 aactgtttct gcccctgacc tgaagagtgt tcgctccaag gtcggctcta cagaaaacat
     6361 caaacaccag cctggaggag gccgggccaa agtagagaaa aaaacagagg cagctaccac
     6421 agctgggaag cctgaaccta atgcagtcac taaagcagcc ggctccattg cgagtgcaca
     6481 gaaaccgcct gctgggaaag tccagatagt atccaaaaaa gtgagctaca gccatattca
     6541 atccaagtgt ggttccaagg acaatattaa gcatgtccct ggaggtggca atgttcagat
     6601 tcagaacaag aaagtggaca tatccaaggt ctcctccaag tgtgggtcca aagctaatat
     6661 caagcacaag cctggtggag gagatgtcaa gattgaaagt cagaagttga acttcaagga
     6721 gaaggcccaa gccaaagtgg gatcccttga taacgtgggc cacttgcctg caggaggtgc
     6781 cgtgaagact gagggcggtg gcagtgaggc ccttccgtgt ccaggccccc ccgctgggga
     6841 ggagccagtc atccctgagg ctgcgcctga cgctggcgcc cctacttcag ccagtggcct
     6901 cagtggccac accaccctgt cagggggtgg tgaccaaagg gagccccaga ccttggacag
     6961 ccagatccag gagacaagca tctaatgatg acattctggt ctcgtccgcg ggcccgggat
     7021 cctgtacaag taaagcggcc gctctagata aatgatagct tgatctggtt accactaaac
     7081 cagcctcaag aacacccgaa tggagtctct aagctacata ataccaactt acactttaca
     7141 aaatgttgtc ccccaaaatg tagccattcg tatctgctcc taataaaaag aaagtttctt
     7201 cacatt
//
