PCDH-TetO-mEGFP-TRIM49-TurboID

LOCUS       PCDH-TetO-mEGFP-TRIM49-TurboID           9545 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 9545)
  AUTHORS   15129015045
  TITLE     Direct Submission
  JOURNAL   Exported Aug 12, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..9545
                     /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..4552
                     /label=trim49
     CDS             4571..4597
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4601..4627
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4631..4657
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4658..5638
                     /codon_start=1
                     /label=TurboID
                     /translation="ASKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLR
                     DWGVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKS
                     GDACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEKEFSRAD"
     misc_feature    5642..6229
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6113..6124)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6302..6535
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6607..6728
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6747..6882
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6920..6936)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6944..6960
                     /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(6968..6998)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7013..7034
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7322..7910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8081..8941)
                     /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(8942..9046)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9520..9536
                     /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 attctggaat cttacaggtc tttcaggggg aactcatctg
     3241 ccccctgtgc atgaactact tcatagaccc ggtcaccata gactgtgggc acagcttttg
     3301 caggccttgt ttctacctca actggcaaga catcccattt cttgtccagt gctctgaatg
     3361 cacaaagtca accgagcaga taaacctcaa aaccaacatt catttgaaga agatggcttc
     3421 tcttgccaga aaagtcagtc tctggctatt cctgagctct gaggagcaaa tgtgtggcac
     3481 tcacagggag acaaagaaga tattctgtga agtggacagg agcctgctct gtttgctgtg
     3541 ctccagctct caggagcacc ggtatcacag acaccgtccc attgagtggg ctgctgagga
     3601 acaccgggag aagcttttac agaaaatgca gtctttgtgg gaaaaagctt gtgaaaatca
     3661 cagaaacctg aatgtggaaa ccaccagaac cagatgctgg aaggattatg tgaatttaag
     3721 gctagaagca attagagctg agtatcagaa gatgcctgca tttcatcatg aagaagaaaa
     3781 acataatttg gagatgctga aaaagaaggg gaaagaaatt tttcatcgac ttcatttaag
     3841 taaagccaaa atggctcata ggatggagat tttaagagga atgtatgagg agctgaacga
     3901 aatgtgccat aaaccagatg tggagctact tcaggctttt ggagacatat tacacaggag
     3961 tgagtccgtg ctgctgcaca tgccccagcc tctgaatcca gagctcagtg cagggcccat
     4021 cactggactg agggacaggc tcaaccaatt ccgagtgcat attactctgc atcatgaaga
     4081 agccaacaat gatatctttc tgtatgaaat tttgagaagc atgtgtattg gatgtgacca
     4141 tcaagatgta ccctatttca ctgcaacacc tagaagtttt cttgcatggg gtgttcagac
     4201 tttcacctcg ggcaaatatt actgggaggt ccatgtaggg gactcctgga attgggcttt
     4261 tggtgtctgt aatatgtatc ggaaagagaa gaatcagaat gagaagatag atggaaaggc
     4321 gggactcttt cttcttgggt gtgttaagaa tgacattcaa tgcagtctct ttaccacctc
     4381 cccacttatg ctgcaatata tcccaaaacc taccagccga gtaggattat tcctggattg
     4441 tgaggctaag actgtgagct ttgttgatgt taatcaaagc tccctaatat acaccatccc
     4501 taattgctct ttctcacctc ctctcaggcc tatcttttgc tgtattcact tcggatccgg
     4561 atctggctcc tacccgtatg atgttccgga ttacgctggc tatccctacg acgtgcccga
     4621 ctatgccggg tacccctatg acgtcccaga ctacgcagct agcaaagaca atactgtgcc
     4681 tctgaagctg atcgctctcc tggctaatgg cgagttccat agtggcgaac agctgggaga
     4741 aaccctgggc atgtccaggg ccgctatcaa caagcacatt cagactctgc gcgactgggg
     4801 cgtggacgtg ttcaccgtgc ccggaaaggg ctactctctg cccgagccta tcccgctgct
     4861 gaacgctaaa cagattctgg gacagctgga cggcgggagc gtggcagtcc tgcctgtggt
     4921 cgactccacc aatcagtacc tgctggatcg aatcggcgag ctgaagagtg gggatgcttg
     4981 cattgcagaa tatcagcagg cagggagagg aagcagaggg aggaaatggt tctctccttt
     5041 tggagctaac ctgtacctga gtatgttttg gcgcctgaag cggggaccag cagcaatcgg
     5101 cctgggcccg gtcatcggaa ttgtcatggc agaagcgctg cgaaagctgg gagcagacaa
     5161 ggtgcgagtc aaatggccca atgacctgta tctgcaggat agaaagctgg caggcatcct
     5221 ggtggagctg gccggaataa caggcgatgc tgcacagatc gtcattggcg ccgggattaa
     5281 cgtggctatg aggcgcgtgg aggaaagcgt ggtcaatcag ggctggatca cactgcagga
     5341 agcagggatt aacctggaca ggaatactct ggccgctacg ctgatccgag agctgcgggc
     5401 agccctggaa ctgttcgagc aggaaggcct ggctccatat ctgccacggt gggagaagct
     5461 ggataacttc atcaatagac ccgtgaagct gatcattggg gacaaagaga ttttcgggat
     5521 tagccggggg attgataaac agggagccct gctgctggaa caggacggag ttatcaaacc
     5581 ctggatgggc ggagaaatca gtctgcggtc tgccgaaaag gagttcagca gggccgactg
     5641 aaatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     5701 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     5761 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     5821 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     5881 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     5941 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     6001 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     6061 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     6121 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     6181 ccgccttcgc cctcagacga gtcggatctc cctttggccg cctccccgcc ggtaccttta
     6241 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     6301 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct
     6361 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     6421 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     6481 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     6541 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     6601 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     6661 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     6721 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt
     6781 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     6841 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg
     6901 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6961 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     7021 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     7081 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     7141 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     7201 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     7261 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7321 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7381 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7441 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7501 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7561 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7621 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7681 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7741 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     7801 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7861 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7921 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7981 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     8041 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     8101 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     8161 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     8221 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     8281 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     8341 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     8401 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8461 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8521 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8581 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8641 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8701 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8761 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8821 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8881 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8941 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     9001 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     9061 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     9121 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     9181 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     9241 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     9301 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     9361 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     9421 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     9481 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca
     9541 agctg
//