LOCUS       sequence-373239-        7621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of shRNA targeting Anillin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7621)
  AUTHORS   Budnar S, Husain KB, Gomez GA, Naghibosadat M, Varma A, Verma S, 
            Hamilton NA, Morris RG, Yap AS
  TITLE     Anillin Promotes Cell Contractility by Cyclic Resetting of RhoA 
            Residence Kinetics
  JOURNAL   Dev Cell. 2019 Jun 17;49(6):894-906.e12. doi: 
            10.1016/j.devcel.2019.04.031. Epub 2019 May 16.
  PUBMED    31105010
REFERENCE   2  (bases 1 to 7621)
  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.devcel.2019.04.031"; journalName: "Dev Cell"; date: 
            "2019-06-17- 17"; volume: "49"; issue: "6"; pages: "894-906.e12"
COMMENT     Sequence Label: pLL5.0-Anillin shRNA
FEATURES             Location/Qualifiers
     source          1..7621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(75..94)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        266..645
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        647..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     800..820
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             863..1043
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1090..1215
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1712..1945
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2130..2174
                     /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    2472..2589
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2644..2956
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2875..2895
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     3019..3035
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3077..3110)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3698..3707
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3704..3706,3707..3709,3710..4423)
                     /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(3704..3706,3707..3709,3710..4420)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3749..3770)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4010..4029)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4357..4378
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4451..4484)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4540..5126
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4593..4613)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5011..5022)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5129..5145)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             5655..5835
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5897..6485)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5977..5996)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6656..7447,7448..7516))
                     /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     7279..7298
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7517..7621)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
       61 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
      121 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
      181 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
      241 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
      301 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      361 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      421 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      481 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      541 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      601 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      661 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      721 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      781 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      841 agcagcgcgt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct
      901 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
      961 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     1021 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
     1081 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
     1141 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
     1201 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
     1261 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
     1321 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
     1381 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     1441 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     1501 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccggc
     1561 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     1621 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     1681 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     1741 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     1801 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     1861 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     1921 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     1981 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     2041 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     2101 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     2161 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     2221 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     2281 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     2341 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
     2401 ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca aatggcagta
     2461 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     2521 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     2581 caaaattttc gggtttatta cagggacagc agagatccag tttggttagt accgcgctct
     2641 agagatccga cgcgccatct ctaggcccgc gccggccccc tcgcacagac ttgtgggaga
     2701 agctcggcta ctcccctgcc ccggttaatt tgcatataat atttcctagt aactatagag
     2761 gcttaatgtg cgataaaaga cagataatct gttcttttta atactagcta cattttacat
     2821 gataggcttg gatttctata agagatacaa atactaaatt attattttaa aaaacagcac
     2881 aaaaggaaac tcaccctaac tgtaaagtaa ttgtgtgttt tgagactata aatatccctt
     2941 ggagaaaagc cttgtttcgg cgatgcctct ttgaataaat tcaagagatt tattcaaaga
     3001 ggcatcgcca ttttttctcg aggtcgacgg tatcgataag ctcgcttcac gagattccag
     3061 caggtcgagg gacctaataa cttcgtatag catacattat acgaagttat attaagggtt
     3121 ccaagcttaa gcggccgctg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa
     3181 cgccattttg caaggcatgg aaaatacata actgagaata gagaagttca gatcaaggtt
     3241 aggaacagag agacagcaga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     3301 cccggctcag ggccaagaac agatggtccc cagatgcggt cccgccctca gcagtttcta
     3361 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     3421 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     3481 taaaagagcc cacaacccct cactcggcgc gccagtcctc cgatagactg cgtcgcccgg
     3541 gtacccgtat tcccaataaa gcctcttgct gtttgcatcc gaatcgtgga ctcgctgatc
     3601 cttgggaggg tctcctcaga ttgattgact gcccacctcg ggggtctttc atcgaattct
     3661 gcagtcgacg gtaccgcggg cccgggatcc accggtcgcc accatggtga gcaagggcga
     3721 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     3781 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     3841 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     3901 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     3961 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     4021 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     4081 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     4141 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     4201 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     4261 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     4321 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     4381 cgccgccggg atcactctcg gcatggacga gctgtacaag taacctgcag gcaattcgtc
     4441 gagggaccta ataacttcgt atagcataca ttatacgaag ttatacatgt ttaagggttc
     4501 cggttccact aggtacaatt cgatatcaag cttatcgata atcaacctct ggattacaaa
     4561 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     4621 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     4681 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     4741 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     4801 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     4861 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     4921 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     4981 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     5041 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     5101 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgatcga gacctagaaa
     5161 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag
     5221 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa
     5281 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     5341 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag
     5401 gctacttccc tgattggcag aactacacac cagggccagg gatcagatat ccactgacct
     5461 ttggatggtg ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag
     5521 gagagaacac ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag
     5581 aagtattaga gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc
     5641 atccggactg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     5701 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     5761 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     5821 ggaaaatctc tagcagcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5881 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5941 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6001 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6061 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6121 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6181 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6241 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6301 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6361 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6421 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6481 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6541 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6601 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6661 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6721 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6781 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6841 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6901 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6961 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7021 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7081 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7141 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7201 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7261 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7321 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7381 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7441 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7501 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7561 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7621 g
//
