LOCUS       sequence-433872-        8101 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of myosin regulatory light chain-DD 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-DD-GFP
FEATURES             Location/Qualifiers
     source          1..8101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(78..97)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        269..648
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        650..848
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     803..823
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             866..1046
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1093..1218
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1715..1948
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2133..2177
                     /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    2475..2592
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2647..2960
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2879..2899
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     2977..2993
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3035..3068)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      4181..4190
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4187..4189,4190..4192,4193..4906)
                     /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(4187..4189,4190..4192,4193..4903)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4232..4253)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4493..4512)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4840..4861
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4934..4967)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5023..5609
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5076..5096)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5494..5505)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5612..5628)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6138..6318
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6380..6968)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6460..6479)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7139..7930,7931..7999))
                     /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     7762..7781
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8000..8101,1..3))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
       61 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
      121 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
      181 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
      241 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
      301 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      361 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      421 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      481 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      541 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      601 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      661 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      721 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      781 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      841 ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga
      901 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
      961 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     1021 ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg
     1081 aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc
     1141 gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga
     1201 gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt
     1261 cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg
     1321 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
     1381 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
     1441 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
     1501 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
     1561 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     1621 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     1681 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     1741 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     1801 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     1861 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     1921 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     1981 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     2041 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     2101 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     2161 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     2221 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     2281 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     2341 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     2401 gatccattcg attagtgaac ggatcggcac tgcgtgcgcc aattctgcag acaaatggca
     2461 gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     2521 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     2581 attcaaaatt ttcgggttta ttacagggac agcagagatc cagtttggtt agtaccgcgc
     2641 tctagagatc cgacgccgcc atctctaggc ccgcgccggc cccctcgcac agacttgtgg
     2701 gagaagctcg gctactcccc tgccccggtt aatttgcata taatatttcc tagtaactat
     2761 agaggcttaa tgtgcgataa aagacagata atctgttctt tttaatacta gctacatttt
     2821 acatgatagg cttggatttc tataagagat acaaatacta aattattatt ttaaaaaaca
     2881 gcacaaaagg aaactcaccc taactgtaaa gtaattgtgt gttttgagac tataaatatc
     2941 ccttggagaa aagccttgtt aacgcgcggt gaccctcgag gtcgacggta tcgataagct
     3001 cgcttcacga gattccagca ggtcgaggga cctaataact tcgtatagca tacattatac
     3061 gaagttatat taagggttcc aagcttaagc ggccgctgaa agaccccacc tgtaggtttg
     3121 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga
     3181 gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaac aaggatatct
     3241 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     3301 cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     3361 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
     3421 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc
     3481 gatagactgc gtcgcccggg tacccgtatt cccaataaag cctcttgctg tttgcatccg
     3541 aatcgtggac tcgctgatcc ttgggagggt ctcctcagat tgattgactg cccacctcgg
     3601 gggtctttca tcgaattctg cagtcgacgg taccgcgggc ccgggatccg ccaccatgtc
     3661 cagcaagcgg gccaaagcca agaccaccaa gaagcggcca cagcgggccg acgacaatgt
     3721 cttcgcaatg tttgaccagt cccagatcca ggagtttaag gaggctttca acatgattga
     3781 ccagaaccgt gatggcttca ttgacaagga ggacctgcac gacatgctgg cctcgctggg
     3841 gaagaacccc acagacgaat acctggaggg catgatgagc gaggccccgg ggcccatcaa
     3901 cttcaccatg ttcctcacca tgtttgggga gaagctgaac ggcacggacc ccgaggatgt
     3961 gattcgcaac gcctttgcct gcttcgacga ggaagcctca ggtttcatcc atgaggacca
     4021 cctccgggag ctgctcacca ccatgggtga ccgcttcaca gatgaggaag tggacgagat
     4081 gtaccgggag gcacccattg ataagaaagg caacttcaac tacgtggagt tcacccgcat
     4141 cctcaaacat ggcgccaagg ataaagacga ccaaccggtc gccaccatgg tgagcaaggg
     4201 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     4261 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     4321 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     4381 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     4441 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     4501 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     4561 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     4621 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     4681 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     4741 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     4801 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     4861 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaacctg caggcaattc
     4921 gtcgagggac ctaataactt cgtatagcat acattatacg aagttataca tgtttaaggg
     4981 ttccggttcc actaggtaca attcgatatc aagcttatcg ataatcaacc tctggattac
     5041 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     5101 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     5161 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     5221 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     5281 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     5341 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     5401 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     5461 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     5521 tcccggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     5581 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgat cgagacctag
     5641 aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc
     5701 tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac
     5761 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     5821 aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac
     5881 aaggctactt ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga
     5941 cctttggatg gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg
     6001 aaggagagaa cacccgcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
     6061 gagaagtatt agagtggagg tttgacagcc gcctagcatt tcatcacatg gcccgagagc
     6121 tgcatccgga ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     6181 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     6241 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     6301 tgtggaaaat ctctagcagc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6361 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6421 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6481 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6541 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6601 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6661 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6721 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6781 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6841 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6901 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6961 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7021 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7081 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7141 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7201 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7261 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7321 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7381 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7441 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7501 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7561 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7621 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7681 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7741 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7801 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7861 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7921 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7981 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8041 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8101 c
//
