LOCUS       sequence-433868-        8101 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of myosin regulatory light chain-AA mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8101)
  AUTHORS   Wee K, Hediyeh-Zadeh S, Duszyc K, Verma S, N Nanavati B, Khare S, 
            Varma A, Daly RJ, Yap AS, Davis MJ, Budnar S
  TITLE     Snail induces epithelial cell extrusion by regulating RhoA 
            contractile signalling and cell-matrix adhesion.
  JOURNAL   J Cell Sci. 2020 Jul 6;133(13). pii: jcs.235622. doi: 
            10.1242/jcs.235622.
  PUBMED    32467325
REFERENCE   2  (bases 1 to 8101)
  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: "J Cell 
            Sci."; date: "2020-07-6"; volume: "133(13). pii: jcs.235622. doi"; 
            pages: " 10.1242/jcs.235622"
COMMENT     Sequence Label: pLL5.0-MRLC-AA-GFP
FEATURES             Location/Qualifiers
     source          1..8101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1684..1917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2102..2146
                     /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    2444..2561
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2616..2929
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2848..2868
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     2946..2962
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3004..3037)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      4150..4159
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4156..4158,4159..4161,4162..4875)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4156..4158,4159..4161,4162..4872)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4201..4222)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4462..4481)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4809..4830
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4903..4936)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4992..5578
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5045..5065)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5463..5474)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5581..5597)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6107..6287
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6349..6937)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6429..6448)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7108..7899,7900..7968))
                     /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     7731..7750
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7969..8073)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
     1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     2581 gcagagatcc agtttggtta gtaccgcgct ctagagatcc gacgccgcca tctctaggcc
     2641 cgcgccggcc ccctcgcaca gacttgtggg agaagctcgg ctactcccct gccccggtta
     2701 atttgcatat aatatttcct agtaactata gaggcttaat gtgcgataaa agacagataa
     2761 tctgttcttt ttaatactag ctacatttta catgataggc ttggatttct ataagagata
     2821 caaatactaa attattattt taaaaaacag cacaaaagga aactcaccct aactgtaaag
     2881 taattgtgtg ttttgagact ataaatatcc cttggagaaa agccttgtta acgcgcggtg
     2941 accctcgagg tcgacggtat cgataagctc gcttcacgag attccagcag gtcgagggac
     3001 ctaataactt cgtatagcat acattatacg aagttatatt aagggttcca agcttaagcg
     3061 gccgctgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa
     3121 ggcatggaaa atacataact gagaatagag aagttcagat caaggttagg aacagagaga
     3181 cagcagaata tgggccaaca aggatatctg tggtaagcag ttcctgcccc ggctcagggc
     3241 caagaacaga tggtccccag atgcggtccc gccctcagca gtttctagag aaccatcaga
     3301 tgtttccagg gtgccccaag gacctgaaaa tgaccctgtg ccttatttga actaaccaat
     3361 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     3421 caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtattc
     3481 ccaataaagc ctcttgctgt ttgcatccga atcgtggact cgctgatcct tgggagggtc
     3541 tcctcagatt gattgactgc ccacctcggg ggtctttcat cgaattctgc agtcgacggt
     3601 accgcgggcc cgggatccgc caccatgtcc agcaagcggg ccaaagccaa gaccaccaag
     3661 aagcggccac agcgggccgc cgccaatgtc ttcgcaatgt ttgaccagtc ccagatccag
     3721 gagtttaagg aggctttcaa catgattgac cagaaccgtg atggcttcat tgacaaggag
     3781 gacctgcacg acatgctggc ctcgctgggg aagaacccca cagacgaata cctggagggc
     3841 atgatgagcg aggccccggg gcccatcaac ttcaccatgt tcctcaccat gtttggggag
     3901 aagctgaacg gcacggaccc cgaggatgtg attcgcaacg cctttgcctg cttcgacgag
     3961 gaagcctcag gtttcatcca tgaggaccac ctccgggagc tgctcaccac catgggtgac
     4021 cgcttcacag atgaggaagt ggacgagatg taccgggagg cacccattga taagaaaggc
     4081 aacttcaact acgtggagtt cacccgcatc ctcaaacatg gcgccaagga taaagacgac
     4141 caaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     4201 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     4261 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     4321 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     4381 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     4441 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     4501 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     4561 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     4621 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     4681 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     4741 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     4801 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     4861 gagctgtaca agtaacctgc aggcaattcg tcgagggacc taataacttc gtatagcata
     4921 cattatacga agttatacat gtttaagggt tccggttcca ctaggtacaa ttcgatatca
     4981 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     5041 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     5101 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     5161 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     5221 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     5281 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     5341 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     5401 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     5461 cttcggccct caatccagcg gaccttcctt cccggcctgc tgccggctct gcggcctctt
     5521 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     5581 cgataccgtc gacctcgatc gagacctaga aaaacatgga gcaatcacaa gtagcaatac
     5641 agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt
     5701 tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     5761 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     5821 tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc agaactacac
     5881 accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt
     5941 tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt
     6001 gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg
     6061 cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt
     6121 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     6181 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     6241 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagca tgtgagcaaa
     6301 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6361 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6421 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6481 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6541 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6601 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6661 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6721 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6781 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6841 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6901 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6961 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7021 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7081 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7141 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7201 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7261 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7321 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7381 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7441 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7501 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7561 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7621 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7681 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7741 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7801 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7861 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7921 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7981 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8041 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8101 c
//
