LOCUS       sequence-327809-        8345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral plasmid with Puromycin selection for expression of 
            mCherry with a C-terminal CAAX polybasic sequence from KRras 
            targeting mCherry to the plasma membrane.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8345)
  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 8345)
  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 PuroR
FEATURES             Location/Qualifiers
     source          1..8345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    54..179
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    676..909
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1094..1138
                     /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    1436..1553
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        1672..2051
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2053..2251
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             2287..2994
                     /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(2432..2450)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2700..2719
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3120..3143)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3470..3489
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3558..4157
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3558..3577)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4054..4074
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4226..4814
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4279..4299)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4697..4708)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4817..4833)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4818..4834)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5343..5523
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5585..6173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5665..5684)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6344..7135,7136..7204))
                     /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     6967..6986
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7205..7309)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7384..7403)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7575..7954
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7956..8154
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             join(8172..8345,1..7)
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
       61 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
      121 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
      181 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
      241 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
      301 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
      361 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
      421 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
      481 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cggccgctga tcttcagacc
      541 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
      601 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
      661 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
      721 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
      781 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
      841 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
      901 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg
      961 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga
     1021 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
     1081 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg
     1141 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg
     1201 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca
     1261 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc
     1321 cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa
     1381 cggatcggca ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta
     1441 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa
     1501 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt
     1561 attacaggga cagcagagat ccagtttggt tagtaccggg cccgggtcga ctagtgatca
     1621 gtgtgaggga gtgtaaagct ggtttcgatg tacgggccag atatacgcgt tgacattgat
     1681 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     1741 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     1801 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     1861 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     1921 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     1981 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     2041 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     2101 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     2161 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     2221 ggcgtgtacg gtgggaggtc tatataagca gcgcgtctaa ctcgaacgct agctgtgcga
     2281 tcgtttatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     2341 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     2401 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     2461 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     2521 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     2581 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     2641 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     2701 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     2761 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     2821 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     2881 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     2941 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caaggaattc
     3001 aagaaaaaga agaagaagtc aaaaaccaag tgtgttatca tgtaagaccc gacattagcg
     3061 ctacagctta agcggaattc taccgggtag gggaggcgct tttcccaagg cagtctggag
     3121 catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca
     3181 cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc
     3241 caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca
     3301 ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg
     3361 agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt
     3421 tctgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg
     3481 cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc
     3541 tgcagcccaa gcttaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     3601 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     3661 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     3721 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     3781 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     3841 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     3901 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     3961 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     4021 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     4081 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     4141 gcaagcccgg tgcctgaccg cgtctggaac aataaacgtt gttgtttggg gttgaattac
     4201 tcttctagac cgcggcgcgc ccgataatca acctctggat tacaaaattt gtgaaagatt
     4261 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     4321 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     4381 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     4441 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     4501 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     4561 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     4621 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     4681 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     4741 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     4801 ggccgcctcc ccgcatcgat accgtcgacc tcgatcgaga cctagaaaaa catggagcaa
     4861 tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg
     4921 aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg
     4981 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     5041 cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg
     5101 attggcagaa ctacacacca gggccaggga tcagatatcc actgaccttt ggatggtgct
     5161 acaagctagt accagttgag caagagaagg tagaagaagc caatgaagga gagaacaccc
     5221 gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtattagagt
     5281 ggaggtttga cagccgccta gcatttcatc acatggcccg agagctgcat ccggactgta
     5341 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     5401 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     5461 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     5521 gcagcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5581 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5641 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5701 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5761 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5821 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5881 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5941 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6001 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6061 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6121 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6181 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6241 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6301 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6361 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6421 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6481 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6541 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6601 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6661 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6721 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6781 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6841 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6901 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6961 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7021 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7081 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7141 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7201 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7261 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7321 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     7381 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     7441 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     7501 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     7561 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     7621 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     7681 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     7741 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     7801 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     7861 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     7921 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     7981 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     8041 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     8101 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt
     8161 ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     8221 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     8281 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     8341 aaatc
//
