PCDH-TetO-mEGFP-TRIM54-TurboID

LOCUS       PCDH-TetO-mEGFP-TRIM54-TurboID             9251 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 9251)
  AUTHORS   15129015045
  TITLE     Direct Submission
  JOURNAL   Exported Aug 13, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..9251
                     /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..4270
                     /label=TRIM54
     CDS             4277..4303
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4307..4333
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4337..4363
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4364..5344
                     /codon_start=1
                     /label=TurboID
                     /translation="ASKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLR
                     DWGVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKS
                     GDACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEKEFSRAD"
     misc_feature    5348..5935
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(5819..5830)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6008..6241
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6313..6434
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6453..6588
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6626..6642)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6650..6666
                     /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(6674..6704)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6719..6740
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7028..7616)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7787..8647)
                     /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(8648..8752)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9226..9242
                     /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 gaattcatga acttcacagt gggtttcaag ccgctgctag gggatgcaca
     3241 cagcatggac aacctggaga agcagctcat ctgccccatc tgcctggaga tgttctccaa
     3301 accagtggtg atcctgccct gccaacacaa cctgtgccgc aaatgtgcca acgacgtctt
     3361 ccaggcctcg aatcctctat ggcagtcccg gggctccacc actgtgtctt caggaggccg
     3421 tttccgctgc ccatcgtgca ggcatgaggt tgtcctggac agacacggtg tctacggcct
     3481 gcagcgaaac ctgctagtgg agaacattat cgacatttac aagcaggagt catccaggcc
     3541 gctgcactcc aaggctgagc agcacctcat gtgcgaggag catgaagaag agaagatcaa
     3601 tatttactgc ctgagctgtg aggtgcccac ctgctctctc tgcaaggtct tcggtgccca
     3661 caaggactgt gaggtggccc cactgcccac catttacaaa cgccagaaga gtgagctcag
     3721 cgatggcatc gcgatgctgg tggcaggcaa tgaccgcgtg caagcagtga tcacacagat
     3781 ggaggaggtg tgccagacta tcgaggacaa tagccggagg cagaagcagt tgttaaacca
     3841 gaggtttgag agcctgtgcg cagtgctgga ggagcgcaag ggtgagctgc tgcaggcgct
     3901 ggcccgggag caagaggaga agctgcagcg cgtccgcggc ctcatccgtc agtatggcga
     3961 ccacctggag gcctcctcta agctggtgga gtctgccatc cagtccatgg aagagccaca
     4021 aatggcgctg tatctccagc aggccaagga gctgatcaat aaggtcgggg ccatgtcgaa
     4081 ggtggagctg gcagggcggc cggagccagg ctatgagagc atggagcaat tcaccgtaag
     4141 ggtggagcac gtggccgaaa tgctgcggac catcgacttc cagccaggcg cttccgggga
     4201 ggaagaggag gtggccccag acggagagga gggcagcgcg gggccggagg aagagcggcc
     4261 ggatgggcct ggatcctacc cgtatgatgt tccggattac gctggctatc cctacgacgt
     4321 gcccgactat gccgggtacc cctatgacgt cccagactac gcagctagca aagacaatac
     4381 tgtgcctctg aagctgatcg ctctcctggc taatggcgag ttccatagtg gcgaacagct
     4441 gggagaaacc ctgggcatgt ccagggccgc tatcaacaag cacattcaga ctctgcgcga
     4501 ctggggcgtg gacgtgttca ccgtgcccgg aaagggctac tctctgcccg agcctatccc
     4561 gctgctgaac gctaaacaga ttctgggaca gctggacggc gggagcgtgg cagtcctgcc
     4621 tgtggtcgac tccaccaatc agtacctgct ggatcgaatc ggcgagctga agagtgggga
     4681 tgcttgcatt gcagaatatc agcaggcagg gagaggaagc agagggagga aatggttctc
     4741 tccttttgga gctaacctgt acctgagtat gttttggcgc ctgaagcggg gaccagcagc
     4801 aatcggcctg ggcccggtca tcggaattgt catggcagaa gcgctgcgaa agctgggagc
     4861 agacaaggtg cgagtcaaat ggcccaatga cctgtatctg caggatagaa agctggcagg
     4921 catcctggtg gagctggccg gaataacagg cgatgctgca cagatcgtca ttggcgccgg
     4981 gattaacgtg gctatgaggc gcgtggagga aagcgtggtc aatcagggct ggatcacact
     5041 gcaggaagca gggattaacc tggacaggaa tactctggcc gctacgctga tccgagagct
     5101 gcgggcagcc ctggaactgt tcgagcagga aggcctggct ccatatctgc cacggtggga
     5161 gaagctggat aacttcatca atagacccgt gaagctgatc attggggaca aagagatttt
     5221 cgggattagc cgggggattg ataaacaggg agccctgctg ctggaacagg acggagttat
     5281 caaaccctgg atgggcggag aaatcagtct gcggtctgcc gaaaaggagt tcagcagggc
     5341 cgactgaaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     5401 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     5461 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     5521 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     5581 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     5641 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     5701 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     5761 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     5821 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     5881 gcgtctccgc cttcgccctc agacgagtcg gatctccctt tggccgcctc cccgccggta
     5941 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     6001 gggggactgg aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact
     6061 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     6121 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     6181 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     6241 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     6301 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6361 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6421 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg
     6481 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     6541 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca
     6601 gagacggccc aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6661 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6721 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6781 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6841 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6901 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6961 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7021 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7081 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7141 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7201 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7261 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7321 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7381 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7441 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     7501 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7561 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7621 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7681 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7741 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7801 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7861 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7921 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7981 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     8041 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8101 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8161 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8221 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8281 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8341 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8401 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8461 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8521 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8581 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8641 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8701 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8761 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     8821 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     8881 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     8941 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     9001 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     9061 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     9121 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     9181 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     9241 gtgccaagct g
//