PCDH-TetO-mEGFP-TRIM74-TurboID

LOCUS       PCDH-TetO-mEGFP-TRIM74-TurboID           8939 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 8939)
  AUTHORS   15129015045
  TITLE     Direct Submission
  JOURNAL   Exported Aug 13, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8939
                     /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..3946
                     /label=TRIM74
     CDS             3965..3991
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3995..4021
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4025..4051
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4052..5032
                     /codon_start=1
                     /label=TurboID
                     /translation="ASKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLR
                     DWGVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKS
                     GDACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEKEFSRAD"
     misc_feature    5036..5623
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(5507..5518)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5696..5929
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6001..6122
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6141..6276
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6314..6330)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6338..6354
                     /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(6362..6392)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6407..6428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6716..7304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7475..8335)
                     /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(8336..8440)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8914..8930
                     /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 gaattcatgg cttggcaggt gagcctgctg gagctggagg actggcttca
     3241 gtgtcccatc tgcctggagg tcttcaagga gtccctaatg ctacagtgcg gccactccta
     3301 ctgcaagggc tgcctggttt ccctgtccta ccacctggac accaaggtgc gctgccccat
     3361 gtgctggcag gtggtggacg gcagcagctc cttgcccaac gtctccctgg cctgggtgat
     3421 cgaagccctg aggctccctg gggacccgga gcccaaggtc tgcgtgcacc accggaaccc
     3481 gctcagcctt ttctgcgaga aggaccagga gctcatctgt ggcctctgcg gtctgctggg
     3541 ctcccaccaa caccacccgg tcacgcccgt ctccaccgtc tgcagccgca tgaaggagga
     3601 gctcgcagcc ctcttctctg agctgaagca ggagcagaag aaggtggatg agctcatcgc
     3661 caaactggtg aaaaaccgga cccgaatcgt caatgagtcg gatgtcttca gctgggtgat
     3721 ccgccgcgag ttccaggagc tgcgccaccc ggtggacgag gagaaggccc gctgcctgga
     3781 ggggataggg ggtcacaccc gtggcctggt ggcctccctg gacatgcagc tggagcaggc
     3841 ccagggaacc cgggagcggc tggcccaagc cgagtgtgtg ctggaacagt tcggaaatga
     3901 ggaccaccat gagttcatct ggaagttcca ctccatggcc tccaggggat ccggatctgg
     3961 ctcctacccg tatgatgttc cggattacgc tggctatccc tacgacgtgc ccgactatgc
     4021 cgggtacccc tatgacgtcc cagactacgc agctagcaaa gacaatactg tgcctctgaa
     4081 gctgatcgct ctcctggcta atggcgagtt ccatagtggc gaacagctgg gagaaaccct
     4141 gggcatgtcc agggccgcta tcaacaagca cattcagact ctgcgcgact ggggcgtgga
     4201 cgtgttcacc gtgcccggaa agggctactc tctgcccgag cctatcccgc tgctgaacgc
     4261 taaacagatt ctgggacagc tggacggcgg gagcgtggca gtcctgcctg tggtcgactc
     4321 caccaatcag tacctgctgg atcgaatcgg cgagctgaag agtggggatg cttgcattgc
     4381 agaatatcag caggcaggga gaggaagcag agggaggaaa tggttctctc cttttggagc
     4441 taacctgtac ctgagtatgt tttggcgcct gaagcgggga ccagcagcaa tcggcctggg
     4501 cccggtcatc ggaattgtca tggcagaagc gctgcgaaag ctgggagcag acaaggtgcg
     4561 agtcaaatgg cccaatgacc tgtatctgca ggatagaaag ctggcaggca tcctggtgga
     4621 gctggccgga ataacaggcg atgctgcaca gatcgtcatt ggcgccggga ttaacgtggc
     4681 tatgaggcgc gtggaggaaa gcgtggtcaa tcagggctgg atcacactgc aggaagcagg
     4741 gattaacctg gacaggaata ctctggccgc tacgctgatc cgagagctgc gggcagccct
     4801 ggaactgttc gagcaggaag gcctggctcc atatctgcca cggtgggaga agctggataa
     4861 cttcatcaat agacccgtga agctgatcat tggggacaaa gagattttcg ggattagccg
     4921 ggggattgat aaacagggag ccctgctgct ggaacaggac ggagttatca aaccctggat
     4981 gggcggagaa atcagtctgc ggtctgccga aaaggagttc agcagggccg actgaaatca
     5041 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     5101 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     5161 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     5221 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     5281 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     5341 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     5401 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     5461 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     5521 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtctccgcct
     5581 tcgccctcag acgagtcgga tctccctttg gccgcctccc cgccggtacc tttaagacca
     5641 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     5701 gggctaattc actcccaacg aaaataagat ctgctttttg cttgtactgg gtctctctgg
     5761 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     5821 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     5881 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
     5941 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     6001 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6061 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6121 tatcatgtct ggctctagct atcccgcccc taactccgcc cagttccgcc cattctccgc
     6181 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     6241 tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga gacggcccaa
     6301 attcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6361 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6421 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6481 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6541 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6601 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6661 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6721 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6781 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6841 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6901 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6961 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7021 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7081 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7141 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7201 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     7261 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     7321 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7381 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     7441 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7501 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7561 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     7621 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7681 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7741 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7801 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7861 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7921 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7981 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     8041 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     8101 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     8161 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     8221 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     8281 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     8341 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     8401 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     8461 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     8521 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     8581 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     8641 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     8701 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     8761 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     8821 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     8881 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagctg
//