LOCUS       sequence-481758-        7361 bp    DNA     circular SYN 08-AUG-2025
DEFINITION  Lentiviral expression of mNeonGreen-tagged PAK1 p21-binding domain 
            (PBD).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7361)
  AUTHORS   Weiss BG, Keth JM, Bhatt K, Doyal M, Hahn KM, Noh J, Isogai T, 
            Danuser G
  TITLE     Morphological control of merlin-Rac antagonism in 
            proliferation-promoting signaling.
  JOURNAL   Sci Signal. 2025 May 20;18(887):eadk0922. doi: 
            10.1126/scisignal.adk0922. Epub 2025 May 20.
  PUBMED    40392939
REFERENCE   2  (bases 1 to 7361)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci 
            Signal."; date: "2025-05-20"; volume: "18(887):eadk0922. doi"; 
            pages: " 10.1126/scisignal.adk0922. Epub 2025 May 20"
COMMENT     Sequence Label: pLVXCMV100-mNeonGreen-PBD
FEATURES             Location/Qualifiers
     source          1..7361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     LTR             1119..1752
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    1959..2547
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2065..2076
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2474..2494
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2862..3569)
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      3305..3314
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3566..3575
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3734..3849
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     CDS             complement(4112..4156)
                     /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    4341..4574
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    5071..5196
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5243..5876
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    6044..6178
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6068..6087)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6122..6141
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        6226..6330
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6331..6399,6400..7191)
                     /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     complement(6549..6568)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
     1021 cgacggtaaa cgctagcaat gtcaaggcct ctcactctct gatattcatt tctttgcaag
     1081 ttataaatac tgaataataa gatgacatga actactactg ctagagattt tccacactga
     1141 ctaaaagggt ctgagggatc tctagttacc agagtcacac aacagacggg cacacactac
     1201 ttgaagcact caaggcaagc tttattgagg cttaagcagt gggttcccta gttagccaga
     1261 gagctcccag gctcagatct ggtctaacca gagagaccca gtacaggcaa aaagcagctg
     1321 cttatatgca ggatctgagg gctcgccact ccccagtccc gcccaggcca cgcctccctg
     1381 gaaagtcccc agcggaaagt cccttgtagc aagctcgata tcagcagttc ttgaagtact
     1441 ccggatgcag ctctcgggcc acgtgatgaa atgctaggcg gctgtcaaac ctccactcta
     1501 acacttctct ctccgggtca tccatcccat gcaggctcac agggtgtaac aagctggtgt
     1561 tctctccttt attggcctct tctaccttat ctggctcaac tggtactagc ttgtagcacc
     1621 atccaaaggt cagtggatat ctgacccctg gccctggtgt gtagttctgc taatcaggga
     1681 agtagccttg tgtgtggtag atccacagat caaggatatc ttgtcttcgt tgggagtgaa
     1741 ttagcccttc cagtcccctc ttttctttta aaaagtggct aagatctaca gctgccttgt
     1801 aagtcattgg tcttaaaggt acctgaggtg tgactggaaa acccacctcc tcctcctctt
     1861 gtgcttctag ccaggcacaa tcagcattgg tagctgctgt attgctactt gtgattgctc
     1921 catgtttttc taggtctcga ctgcagaatt aattccaggc ggggaggcgg cccaaaggga
     1981 gatccgactc gtctgagggc gaaggcgaag acgcggaaga ggccgcagag ccggcagcag
     2041 gccgcgggaa ggaaggtccg ctggattgag ggccgaaggg acgtagcaga aggacgtccc
     2101 gcgcagaatc caggtggcaa cacaggcgag cagccatgga aaggacgtca gcttccccga
     2161 caacaccacg gaattgtcag tgcccaacag ccgagcccct gtccagcagc gggcaaggca
     2221 ggcggcgatg agttccgccg tggcaatagg gagggggaaa gcgaaagtcc cggaaaggag
     2281 ctgacaggtg gtggcaatgc cccaaccagt gggggttgcg tcagcaaaca cagtgcacac
     2341 cacgccacgt tgcctgacaa cgggccacaa ctcctcataa agagacagca accaggattt
     2401 atacaaggag gagaaaatga aagccatacg ggaagcaata gcatgataca aaggcattaa
     2461 agcagcgtat ccacatagcg taaaaggagc aacatagtta agaataccag tcaatctttc
     2521 acaaattttg taatccagag gttgattgtt ccagacgcgt cgggccctct agactcgagt
     2581 tatttctggc tgttggatgt cttcttcgag ttgtaaaact ccaacacatc cagaacagcc
     2641 tgcgggtttt tcttctgctc cgacttagtg atatttgatg tctgaagcaa gcgggcccac
     2701 tgctctggca ttcccgtaaa ctcccctgtg acagcatcaa aaccgacatg aattgtgtgt
     2761 tcaaaatctg aagggagaga aatctctggc cgctctttct ctttcttttt atttgtttta
     2821 tctccaggta aaatggatcc gagctcagat ctgagtccgg acttgtacag ctcgtccatg
     2881 cccatcacat cggtaaaggc cttttgccac tccttgaagt tgagctcggt cttggagtgc
     2941 ttgagctccg tcttacggaa cacgtacatc ggctggttct tcagatagtt agccgccatt
     3001 ggcttggcaa aggtgtaggt ggtccgcgca gtgctccggt agcgcttgcc atttccagtg
     3061 gtgtaactcc acttaaaggt actgatgatg gttttgtcgt tggggtaagt cttcttcgac
     3121 ctgcaccagt ccgcagcggt cagcgagttg gtcatcacag gaccgtcagc agggaaacca
     3181 gtccccttca cctgggcctc tcctttgatg tggcttccct cgtaggtgta gcggtagtta
     3241 acagtaaggg aggcaccatc ttcaaactgc attgtgcgat ggacttggta gccggagcca
     3301 tctaccatgg cggcctggaa aggcgacatc ccgtcagggt agggcaggta ctgatggaag
     3361 ccatacccga tatgagggac cagaatccag ggggagaact ggaggtcacc cttggtggac
     3421 ttcaggttta actcctcata accatcattt ggattgccgg tgccctgacc caccatgtca
     3481 aagtccacac cgttgatgga gccaaagatg tgtaactcat gtgtcgctgg gagagaggcc
     3541 atgttatcct cctcgccctt gctcaccatg gtggcactag ttagcgctag cggcggtcag
     3601 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
     3661 aatgaccccg taattgatta ctattaataa ctatcgataa actggatctc tgctgtccct
     3721 gtaataaacc cgaaaatttt gaatttttgt aatttgtttt tgtaattctt tagtttgtat
     3781 gtctgttgct attatgtcta ctattctttc ccctgcactg taccccccaa tccccccttt
     3841 tcttttaaag gcgataccgt cgagatccgt tcactaatcg aatggatctg tctctgtctc
     3901 tctctccacc ttcttcttct attccttcgg gcctgtcggg tcccctcggg gttgggaggt
     3961 gggtctgaaa cgataatggt gaatatccct gcctaactct attcactata gaaagtacag
     4021 caaaaactat tcttaaacct accaagcctc ctactatcat tatgaataat tttatatacc
     4081 acagccaatt tgttatgtta aaccaattcc acaaacttgc ccatttatct aattccaata
     4141 attcttgttc attcttttct tgctggtttt gcgattcttc aattaaggag tgtattaagc
     4201 ttgtgtaatt gttaatttct ctgtcccact ccatccaggt cgtgtgattc caaatctgtt
     4261 ccagagattt attactccaa ctagcattcc aaggcacagc agtggtgcaa atgagttttc
     4321 cagagcaacc ccaaatcccc aggagctgtt gatcctttag gtatctttcc acagccagga
     4381 ttcttgcctg gagctgcttg atgccccaga ctgtgagttg caacagatgc tgttgcgcct
     4441 caatagccct cagcaaattg ttctgctgct gcactatacc agacaataat tgtctggcct
     4501 gtaccgtcag cgtcattgac gctgcgccca tagtgcttcc tgctgctccc aagaacccaa
     4561 ggaacaaagc tcctattccc actgctcttt tttctctctg caccactctt ctctttgcct
     4621 tggtgggtgc tactcctaat ggttcaattt ttactacttt atatttatat aattcacttc
     4681 tccaattgtc cctcatatct cctcctccag gtctgaagat cagcggccgg ccgcttgctg
     4741 tgcggtggtc ttacttttgt tttgctcttc ctctatcttg tctaaagctt ccttggtgtc
     4801 ttttatctct atcctttgat gcacacaata gagggttgct actgtattat ataatgatct
     4861 aagttcttct gatcctgtct gaagggatgg ttgtagctgt cccagtattt gtctacagcc
     4921 ttctgatgtt tctaacaggc caggattaac tgcgaatcgt tctagctccc tgcttgccca
     4981 tactatatgt tttaatttat attttttctt tccccctggc cttaaccgaa ttttttccca
     5041 tcgcgatcta attctccccc gcttaatact gacgctctcg cacccatctc tctccttcta
     5101 gcctccgcta gtcaaaattt ttggcgtact caccagtcgc cgcccctcgc ctcttgccgt
     5161 gcgcgcttca gcaagccgag tcctgcgtcg agagagctcc tctggtttcc ctttcgcttt
     5221 caagtccctg ttcgggcgcc actgctagag attttccaca ctgactaaaa gggtctgagg
     5281 gatctctagt taccagagtc acacaacaga cgggcacaca ctacttgaag cactcaaggc
     5341 aagctttatt gaggcttaag cagtgggttc cctagttagc cagagagctc ccaggctcag
     5401 atctggtcta accagagaga cccagtacag gcaaaaagca gctgcttata tgcaggatct
     5461 gagggctcgc cactccccag tcccgcccag gccacgcctc cctggaaagt ccccagcgga
     5521 aagtcccttg tagcaagctc gatatcagca gttcttgaag tactccggat gcagctctcg
     5581 ggccacgtga tgaaatgcta ggcggctgtc aaacctccac tctaacactt ctctctccgg
     5641 gtcatccatc ccatgcaggc tcacagggtg taacaagctg gtgttctctc ctttattggc
     5701 ctcttctacc ttatctggct caactggtac tagcttgtag caccatccaa aggtcagtgg
     5761 atatctgacc cctggccctg gtgtgtagtt ctgctaatca gggaagtagc cttgtgtgtg
     5821 gtagatccac agatcaagga tatcttgtct tctttgggag tgaattagcc cttccaacta
     5881 ctaagtttgt agtacatatt taacaaatac aatttcttta aaatgaaaat aattcagagg
     5941 aatcacaggt ttagagtaaa tgaaaccaca ggtaattggc agtggtaata gggtatgggg
     6001 tgggaagttt gggatgattt tggttagctt gagttatcca gttgatccag acatgataag
     6061 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     6121 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttga
     6181 tctcccgatc cgtcgacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6241 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6301 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6361 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6421 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6481 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6541 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6601 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6661 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6721 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6781 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6841 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6901 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6961 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     7021 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     7081 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     7141 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     7201 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7261 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7321 cactgagcgt cagaccccgt agaaaagatc aaaggatctt c
//
