PCDH-CMV-FRB-mEGFP-TBK1

LOCUS       PCDH-CMV-FRB-mEGFP-TBK1             9477 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 9477)
  AUTHORS   15129015045
  TITLE     Direct Submission
  JOURNAL   Exported Aug 13, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..9477
                     /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"
     enhancer        1938..2241
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2242..2445
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    2499..2591
                     /label=iZF
     CDS             2628..3341
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     regulatory      2628
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3342..3371
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    5574..6161
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6045..6056)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6234..6467
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6539..6660
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6679..6814
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6852..6868)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6876..6892
                     /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(6900..6930)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6945..6966
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7254..7842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8013..8873)
                     /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(8874..8978)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9452..9468
                     /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 tactagttgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     1981 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     2041 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     2101 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     2161 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     2221 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     2281 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     2341 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     2401 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
     2461 cagattctag accgctagcg ctaccggtcg ccaccatgac tggagaacgg cccttccagt
     2521 gcaatcagtg cggggcctca ttcacccaga agggcaacct gctccggcac atcaagctgc
     2581 attccgggga gggcggaagt ggaggatctg gaggttcagg tggttctgtg agcaagggcg
     2641 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2701 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2761 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2821 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     2881 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2941 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     3001 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3061 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     3121 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     3181 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     3241 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     3301 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtccggactc agatctcgag
     3361 ctcaagcttc ggaattcatg cagagcactt ctaatcatct gtggctttta tctgatattt
     3421 taggccaagg agctactgca aatgtctttc gtggaagaca taagaaaact ggtgatttat
     3481 ttgctatcaa agtatttaat aacataagct tccttcgtcc agtggatgtt caaatgagag
     3541 aatttgaagt gttgaaaaaa ctcaatcaca aaaatattgt caaattattt gctattgaag
     3601 aggagacaac aacaagacat aaagtactta ttatggaatt ttgtccatgt gggagtttat
     3661 acactgtttt agaagaacct tctaatgcct atggactacc agaatctgaa ttcttaattg
     3721 ttttgcgaga tgtggtgggt ggaatgaatc atctacgaga gaatggtata gtgcaccgtg
     3781 atatcaagcc aggaaatatc atgcgtgtta taggggaaga tggacagtct gtgtacaaac
     3841 tcacagattt tggtgcagct agagaattag aagatgatga gcagtttgtt tctctgtatg
     3901 gcacagaaga atatttgcac cctgatatgt atgagagagc agtgctaaga aaagatcatc
     3961 agaagaaata tggagcaaca gttgatcttt ggagcattgg ggtaacattt taccatgcag
     4021 ctactggatc actgccattt agaccctttg aagggcctcg taggaataaa gaagtgatgt
     4081 ataaaataat tacaggaaag ccttctggtg caatatctgg agtacagaaa gcagaaaatg
     4141 gaccaattga ctggagtgga gacatgcctg tttcttgcag tctttctcgg ggtcttcagg
     4201 ttctacttac ccctgttctt gcaaacatcc ttgaagcaga tcaggaaaag tgttggggtt
     4261 ttgaccagtt ttttgcagaa actagtgata tacttcaccg aatggtaatt catgtttttt
     4321 cgctacaaca aatgacagct cataagattt atattcatag ctataatact gctactatat
     4381 ttcatgaact ggtatataaa caaaccaaaa ttatttcttc aaatcaagaa cttatctacg
     4441 aagggcgacg cttagtctta gaacctggaa ggctggcaca acatttccct aaaactactg
     4501 aggaaaaccc tatatttgta gtaagccggg aacctctgaa taccatagga ttaatatatg
     4561 aaaaaatttc cctccctaaa gtacatccac gttatgattt agacggggat gctagcatgg
     4621 ctaaggcaat aacaggggtt gtgtgttatg cctgcagaat tgccagtacc ttactgcttt
     4681 atcaggaatt aatgcgaaag gggatacgat ggctgattga attaattaaa gatgattaca
     4741 atgaaactgt tcacaaaaag acagaagttg tgatcacatt ggatttctgt atcagaaaca
     4801 ttgaaaaaac tgtgaaagta tatgaaaagt tgatgaagat caacctggaa gcggcagagt
     4861 taggtgaaat ttcagacata cacaccaaat tgttgagact ttccagttct cagggaacaa
     4921 tagaaaccag tcttcaggat atcgacagca gattatctcc aggtggatca ctggcagacg
     4981 catgggcaca tcaagaaggc actcatccga aagacagaaa tgtagaaaaa ctacaagtcc
     5041 tgttaaattg catgacagag atttactatc agttcaaaaa agacaaagca gaacgtagat
     5101 tagcttataa tgaagaacaa atccacaaat ttgataagca aaaactgtat taccatgcca
     5161 caaaagctat gacgcacttt acagatgaat gtgttaaaaa gtatgaggca tttttgaata
     5221 agtcagaaga atggataaga aagatgcttc atcttaggaa acagttatta tcgctgacta
     5281 atcagtgttt tgatattgaa gaagaagtat caaaatatca agaatatact aatgagttac
     5341 aagaaactct gcctcagaaa atgtttacag cttccagtgg aatcaaacat accatgaccc
     5401 caatttatcc aagttctaac acattagtag aaatgactct tggtatgaag aaattaaagg
     5461 aagagatgga aggggtggtt aaagaacttg ctgaaaataa ccacatttta gaaaggtttg
     5521 gctctttaac catggatggt ggccttcgca acgttgactg tctttaggga tccaatcaac
     5581 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     5641 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     5701 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     5761 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     5821 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     5881 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     5941 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     6001 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     6061 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt ctccgccttc
     6121 gccctcagac gagtcggatc tccctttggc cgcctccccg ccggtacctt taagaccaat
     6181 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     6241 gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt ctctctggtt
     6301 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     6361 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     6421 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     6481 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     6541 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6601 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     6661 tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca ttctccgccc
     6721 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     6781 ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga cggcccaaat
     6841 tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     6901 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     6961 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     7021 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     7081 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     7141 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     7201 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     7261 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7321 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7381 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7441 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7501 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7561 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7621 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7681 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7741 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7801 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7861 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7921 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7981 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     8041 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     8101 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     8161 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     8221 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     8281 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     8341 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     8401 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     8461 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     8521 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8581 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8641 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8701 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8761 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8821 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8881 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8941 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     9001 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     9061 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     9121 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     9181 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     9241 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     9301 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     9361 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     9421 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctg
//