LOCUS       sequence-135014-        8175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral plasmid to express destabilized GFP under the control of 
            the ATF-responsive CHOP promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8175)
  AUTHORS   Boitard M, Bocchi R, Egervari K, Petrenko V, Viale B, Gremaud S, 
            Zgraggen E, Salmon P, Kiss JZ
  TITLE     Wnt signaling regulates multipolar-to-bipolar transition of 
            migrating neurons in the cerebral cortex.
  JOURNAL   Cell Rep. 2015 Mar 3;10(8):1349-61. doi: 
            10.1016/j.celrep.2015.01.061. Epub 2015 Feb 26.
  PUBMED    25732825
REFERENCE   2  (bases 1 to 8175)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2015.01.061"; journalName: "Cell Rep"; date: 
            "2015-03-3- 3"; volume: "10"; issue: "8"; pages: "1349-61"
COMMENT     Sequence Label: pCLX-CHOP-dGFP
FEATURES             Location/Qualifiers
     source          1..8175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             385..642
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    689..814
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1306..1423
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    1783..2016
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2201..2245
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     protein_bind    2520..2540
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    3279..3303
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(3309..3311,3312..3314,3315..4025,4026..4031,
                     4032..4154)
                     /codon_start=1
                     /product="EGFP destabilized by fusion to residues 422-461 
                     of mouse ornithine decarboxylase, giving an in vivo 
                     half-life of ~2 hours"
                     /label=d2EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     primer_bind     complement(3354..3375)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3615..3634)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3962..3983
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4155..4179)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4197..4785
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4250..4270)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4668..4679)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(4928..4961)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             4981..5161
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5199..5218)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5264..5319
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5318..5337)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5678..5694)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5678..5694)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5691..5713)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5702..5718
                     /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(join(5726..5732,5733..5750,5751..5756))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5771..5792
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5851..6046
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5897..6032
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6052..6186
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6089..6108)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6143..6162
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6254..6271)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6425..7013)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6505..6524)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7184..7975,7976..8044))
                     /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"
     primer_bind     7807..7826
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8045..8149)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggaggcg tggcctgggc gggactgggg agtggcgagc
      421 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc
      481 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
      541 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
      601 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
      661 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
      721 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
      781 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
      841 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
      901 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
      961 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     1021 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     1081 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaaaaa
     1141 agcacagcaa gcagcaggat cttcagacct ggaaattccc tacaatcccc aaagtcaagg
     1201 agtagtagaa tctatgaata aagaattaaa gaaaattata ggacaggtaa gagatcaggc
     1261 tgaacatctt aagacagcag tacaaatggc agtattcatc cacaatttta aaagaaaagg
     1321 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     1381 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     1441 cagcagaaat ccactttgga aaggaccagc aaagctcctc tggaaaggtg aaggggcagt
     1501 agtaatacaa gataatagtg acataaaagt agtgccaaga agaaaagcaa agatcattag
     1561 ggattatgga aaacagatgg caggtgatga ttgtgtggca agtagacagg atgaggatta
     1621 gaacatggaa aagtttagta aaacaccata aggaggagat atgagggaca attggagaag
     1681 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1741 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1801 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1861 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1921 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1981 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     2041 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     2101 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     2161 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     2221 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     2281 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     2341 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     2401 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     2461 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatctagtga ttatcggatc
     2521 aactttgtat agaaaagttg tcgagatgtc gacaatccca gtggatggat accaacttta
     2581 aaaagaaaag ttcaaaaggc ctatgtgccc attagctggg aggggccaag aaatatggga
     2641 gtcccttata gtgggggtaa aacggcgggt aaagctaggt gggcggaaca gcagcttctg
     2701 ggggagacaa gcggcaaaga ggctcacgac cgactagggg cgaccaaggc tgatagccgt
     2761 tggggccgtt gggcgccggg agctggcgcc ccgccctctc tcctctcccc caccctccgc
     2821 acctcccacc accctcggtg tcccctgcgc gtgcgcgtgc agacaccggt tgccaaacat
     2881 tgcatcatcc ccgcccccct ttcctcccct ccccccccgc tacactcccc tccgcgcgcg
     2941 cgcatgactc acccacctcc tccgtgaagc ctcgtgaccc aaagccactt ccgggtccga
     3001 cactacgtcg accccctagc gagagggagc gacgggggcg gtgccgcggg gctcctgagt
     3061 ggcggatgcg agggacgggg cggggccaat gccggcgtgc cactttctga ttggtaggtt
     3121 ttggggtccc gcccctgaga ggagggcaag gccatggtaa aagattacag ccaggcgctc
     3181 ccgaggtcag agacttaagt ctaaggcact gagcgtatca tgttaaagat gagcgggtgg
     3241 cagcgacaga gccaaaatca gagctggaac ctgaggagac aagtttgtac aaaaaagcag
     3301 gcttcaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     3361 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     3421 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     3481 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     3541 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     3601 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     3661 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     3721 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     3781 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     3841 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     3901 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     3961 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     4021 acaagaagct tagccatggc ttcccgccgg aggtggagga gcaggatgat ggcacgctgc
     4081 ccatgtcttg tgcccaggag agcgggatgg accgtcaccc tgcagcctgt gcttctgcta
     4141 ggatcaatgt gtagacccag ctttcttgta caaagtggtg ataatcgaat tccgataatc
     4201 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     4261 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     4321 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     4381 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     4441 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     4501 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     4561 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     4621 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     4681 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     4741 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcgg gaattcccgc
     4801 ggttcgcttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     4861 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatggg atcaattcac
     4921 catgggaata acttcgtata gcatacatta tacgaagtta tgctgctttt tgcttgtact
     4981 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     5041 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     5101 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     5161 agcatctagt tgaatcgcca gtgtgtcgag ctagcagatc tttttccctc tgccaaaaat
     5221 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     5281 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     5341 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgccata
     5401 tgctggctgc catgaacaaa ggtggctata aagaggtcat cagtatatga aacagccccc
     5461 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     5521 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     5581 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatgaag
     5641 atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5701 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5761 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5821 agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc
     5881 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5941 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     6001 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt
     6061 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     6121 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     6181 tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6241 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6301 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6361 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6421 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6481 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6541 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6601 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     6661 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6721 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6781 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6841 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6901 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6961 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7021 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7081 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7141 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7201 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7261 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7321 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7381 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7441 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7501 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7561 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7621 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7681 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7741 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7801 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7861 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7921 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7981 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8041 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8101 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8161 gaaaagtgcc acctg
//
