LOCUS       sequence-252373-        7222 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral plasmid for expression of fluorescent reporter mCherry 
            targeted to the plasma membrane via CAAX polybasic sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7222)
  AUTHORS   Janska L, Anandi L, Kirchberger NC, Marinkovic ZS, Schachtner LT, 
            Guzelsoy G, Carmona-Fontaine C
  TITLE     The MEMIC is an ex vivo system to model the complexity of the tumor 
            microenvironment.
  JOURNAL   Dis Model Mech. 2021 Aug 1;14(8). pii: 271783. doi: 
            10.1242/dmm.048942. Epub 2021 Aug 18.
  PUBMED    34407185
REFERENCE   2  (bases 1 to 7222)
  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"; journalName: "Dis Model 
            Mech."; date: "2021-08-1"; volume: "14(8). pii: 271783. doi"; pages:
            " 10.1242/dmm.048942. Epub 2021 Aug 18"
COMMENT     Sequence Label: pLenti-mCherry-CAAX
FEATURES             Location/Qualifiers
     source          1..7222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    850..1083
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1268..1312
                     /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"
     misc_feature    1610..1727
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        1897..2200
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2202..2400
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             2436..3143
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2581..2599)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2849..2868
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3277..3865
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3330..3350)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3748..3759)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3868..3884)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3869..3885)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             4394..4574
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4636..5224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4716..4735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5395..6186,6187..6255))
                     /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     6018..6037
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6256..6360)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6435..6454)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6626..7005
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7007..7205
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccggccg ctgatcttca gacctggagg
      721 aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga
      781 accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc
      841 agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc
      901 agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca
      961 gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg
     1021 catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct
     1081 cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc
     1141 tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga
     1201 cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca
     1261 gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg
     1321 gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt
     1381 ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata
     1441 ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg
     1501 aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc
     1561 ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa
     1621 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
     1681 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
     1741 gggacagcag agatccagtt tggttagtac cgggcccggg tcgactagtg atcagtgtga
     1801 gggagtgtaa agctggtttn cgatgtmsgg scmrrwtawm sssttkrmmt trwytattrm
     1861 skrgttatta rtwswwagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     1921 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     1981 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     2041 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     2101 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     2161 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     2221 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     2281 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     2341 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     2401 cgcgtctaac tcgaacgcta gctgtgcgat cgtttatggt gagcaagggc gaggaggata
     2461 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     2521 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     2581 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     2641 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     2701 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     2761 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     2821 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     2881 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     2941 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     3001 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     3061 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     3121 gcggcatgga cgagctgtac aaggaattca agaaaaagaa gaagaagtca aaaaccaagt
     3181 gtgttatcat gtaaaaacgt tgttgtttgg ggttgaayww stmwwmwmgw ygttgtttgg
     3241 ggttgaatta ctcttctaga ccgcggcgcg cccgataatc aacctctgga ttacaaaatt
     3301 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3361 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3421 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3481 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     3541 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     3601 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     3661 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     3721 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3781 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3841 atctcccttt gggccgcctc cccgcatcga taccgtcgac ctcgatcgag acctagaaaa
     3901 acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga
     3961 agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat
     4021 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     4081 gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg
     4141 ctacttccct gattggcaga actacacacc agggccaggg atcagatatc cactgacctt
     4201 tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg
     4261 agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
     4321 agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca
     4381 tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     4441 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     4501 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     4561 gaaaatctct agcagcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4621 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4681 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4741 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4801 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4861 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4921 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4981 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5041 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5101 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5161 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5221 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5281 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5341 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5401 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5461 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5521 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5581 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5641 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5701 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5761 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5821 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5881 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5941 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6001 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6061 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6121 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6181 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6241 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6301 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6361 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     6421 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     6481 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     6541 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     6601 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
     6661 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     6721 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     6781 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     6841 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     6901 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     6961 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     7021 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     7081 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     7141 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     7201 agcagcgcgt tttgcctgta ct
//
