PCDH-TetO-mEGFP-TRIM48-TurboID

LOCUS      PCDH-TetO-mEGFP-TRIM48-TurboID               8861 bp DNA     circular SYN 04-JUL-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8861)
  AUTHORS   15129015045
  TITLE     Direct Submission
  JOURNAL   Exported Aug 12, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             234..414
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    458..583
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1076..1309
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1798..1915
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    1980..2250
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2283..2320
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             2444..3160
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      2444..2447
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3161..3190
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    3197..3868
                     /label=TRIM48
     CDS             3887..3913
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3917..3943
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3947..3973
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3974..4954
                     /codon_start=1
                     /label=TurboID
                     /translation="ASKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLR
                     DWGVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKS
                     GDACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEKEFSRAD"
     misc_feature    4958..5545
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(5429..5440)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5618..5851
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5923..6044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6063..6198
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6236..6252)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6260..6276
                     /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(6284..6314)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6329..6350
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6638..7226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7397..8257)
                     /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"
     promoter        complement(8258..8362)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8836..8852
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtcgg gtttattaca gggacagcag agatccagtt tggttatcga gtttaccact
     1981 ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga
     2041 aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagttt
     2101 accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga
     2161 tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc
     2221 gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagctcgg tacccgggtc
     2281 gagtaggcgt gtacggtggg aggcctatat aagcagagct cgtttagtga accgtcagat
     2341 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
     2401 cctccgcggc ccctctagac cgctagcgct accggtcgcc accatggtga gcaagggcga
     2461 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2521 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2581 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2641 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2701 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     2761 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2821 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     2881 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     2941 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     3001 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     3061 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     3121 cgccgccggg atcactctcg gcatggacga gctgtacaag tccggactca gatctcgagc
     3181 tcaagcttcg gaattcatgt ctcgaagaat cattgtggga acccttcaaa gaacccagcg
     3241 aaacatgaat tctggaatct cgcaagtctt ccagagggaa ctcacctgcc ccatctgcat
     3301 gaactacttc atagacccgg tcaccataga ctgtgggcac agcttttgca ggccctgttt
     3361 ctacctcaac tggcaagaca tcccaattct tactcagtgc tttgaatgca taaagacaat
     3421 acagcagaga aacctcaaaa ctaacattcg attgaagaag atggcttccc ttgccagaaa
     3481 agccagtctc tggctattcc tgagctctga ggagcaaatg tgtggcattc acagggagac
     3541 aaagaagatg ttctgtgaag tggacaggag cctgctctgt ttgctgtgct ccagctctca
     3601 ggagcaccgg tatcacagac actgtcccgc tgagtgggct gctgaggaac actgggagaa
     3661 gcttttaaag aaaatgcagt ctttatggga aaaagcttgt gaaaatcaga gaaacctgaa
     3721 tgtggaaacc accagaatca gccactggaa ggcttttgga gacatattat acaggagtga
     3781 gtccgtgctg ctgcacatgc cccagcctct gaatctagcg ctcagggcag ggcccatcac
     3841 tggactgagg gacaggctca accaattcgg atccggatct ggctcctacc cgtatgatgt
     3901 tccggattac gctggctatc cctacgacgt gcccgactat gccgggtacc cctatgacgt
     3961 cccagactac gcagctagca aagacaatac tgtgcctctg aagctgatcg ctctcctggc
     4021 taatggcgag ttccatagtg gcgaacagct gggagaaacc ctgggcatgt ccagggccgc
     4081 tatcaacaag cacattcaga ctctgcgcga ctggggcgtg gacgtgttca ccgtgcccgg
     4141 aaagggctac tctctgcccg agcctatccc gctgctgaac gctaaacaga ttctgggaca
     4201 gctggacggc gggagcgtgg cagtcctgcc tgtggtcgac tccaccaatc agtacctgct
     4261 ggatcgaatc ggcgagctga agagtgggga tgcttgcatt gcagaatatc agcaggcagg
     4321 gagaggaagc agagggagga aatggttctc tccttttgga gctaacctgt acctgagtat
     4381 gttttggcgc ctgaagcggg gaccagcagc aatcggcctg ggcccggtca tcggaattgt
     4441 catggcagaa gcgctgcgaa agctgggagc agacaaggtg cgagtcaaat ggcccaatga
     4501 cctgtatctg caggatagaa agctggcagg catcctggtg gagctggccg gaataacagg
     4561 cgatgctgca cagatcgtca ttggcgccgg gattaacgtg gctatgaggc gcgtggagga
     4621 aagcgtggtc aatcagggct ggatcacact gcaggaagca gggattaacc tggacaggaa
     4681 tactctggcc gctacgctga tccgagagct gcgggcagcc ctggaactgt tcgagcagga
     4741 aggcctggct ccatatctgc cacggtggga gaagctggat aacttcatca atagacccgt
     4801 gaagctgatc attggggaca aagagatttt cgggattagc cgggggattg ataaacaggg
     4861 agccctgctg ctggaacagg acggagttat caaaccctgg atgggcggag aaatcagtct
     4921 gcggtctgcc gaaaaggagt tcagcagggc cgactgaaat caacctctgg attacaaaat
     4981 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5041 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5101 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5161 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5221 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     5281 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     5341 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     5401 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     5461 cggcctgctg ccggctctgc ggcctcttcc gcgtctccgc cttcgccctc agacgagtcg
     5521 gatctccctt tggccgcctc cccgccggta cctttaagac caatgactta caaggcagct
     5581 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     5641 cgaaaataag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     5701 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     5761 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     5821 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     5881 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     5941 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6001 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     6061 ctatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6121 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     6181 gaggcttttt tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat
     6241 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6301 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6361 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6421 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6481 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6541 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6601 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6661 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6721 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6781 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6841 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6901 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6961 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7021 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     7081 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7141 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7201 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7261 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7321 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7381 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7441 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7501 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7561 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7621 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7681 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7741 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7801 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7861 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7921 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7981 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8041 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8101 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8161 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8221 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8281 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8341 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     8401 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     8461 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     8521 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     8581 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     8641 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     8701 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     8761 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     8821 cccagtcacg acgttgtaaa acgacggcca gtgccaagct g
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