LOCUS       sequence-481769-        8970 bp    DNA     circular SYN 08-AUG-2025
DEFINITION  Lentiviral expression of SNAP-tagged NF2 fragment (aa 1-27).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8970)
  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 8970)
  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: pLVX-IRES-NEO-NF2(1-27)-SNAP-tag
FEATURES             Location/Qualifiers
     source          1..8970
                     /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(2561..3355)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2672..2691)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3282..3301
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     misc_feature    join(3356..3364,3365..3367,3368..3942)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3529..3548)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     3758..3775
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     regulatory      4668..4677
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4757..4781)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(4778..4981)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4807..4827)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4982..5285
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    5343..5458
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     CDS             complement(5721..5765)
                     /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    5950..6183
                     /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    6680..6805
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6852..7485
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    7653..7787
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7677..7696)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7731..7750
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        7835..7939
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7940..8008,8009..8800)
                     /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(8158..8177)
                     /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 tcagaagaac tcgtcaagaa
     2581 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
     2641 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
     2701 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
     2761 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg
     2821 gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
     2881 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat
     2941 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
     3001 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc
     3061 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag
     3121 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt
     3181 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
     3241 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
     3301 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca gccatggttg
     3361 tggcaagctt atcatcgtgt ttttcaaagg aaaaccacgt ccccgtggtt cggggggcct
     3421 agacgttttt ttaacctcga ctaaacacat gtaaagcatg tgcaccgagg ccccagatca
     3481 gatcccatac aatggggtac cttctgggca tccttcagcc ccttgttgaa tacgcttgag
     3541 gagagccatt tgactctttc cacaactatc caactcacaa cgtggcactg gggttgtgcc
     3601 gcctttgcag gtgtatctta tacacgtggc ttttggccgc agaggcacct gtcgccaggt
     3661 ggggggttcc gctgcctgca aagggtcgct acagacgttg tttgtcttca agaagcttcc
     3721 agaggaactg cttccttcac gacattcaac agaccttgca ttcctttggc gagaggggaa
     3781 agacccctag gaatgctcgt caagaagaca gggccaggtt tccgggccct cacattgcca
     3841 aaagacggca atatggtgga aaataacata tagacaaacg cacaccggcc ttattccaag
     3901 cggcttcggc cagtaacgtt aggggggggg gagggagagg ggcggaatcc ttagctaacg
     3961 cgtgaattcg atatccttac ccagcccagg cttgcccagt ctgtggccct cgtgggccag
     4021 cagccattcc tttaccgcga ggcccccctc ataaccacct acgtcgagat caccctgtac
     4081 cactctatga caggggatca gaatgggcac gggatttccg ctcagggcgg ttttcacggc
     4141 ggcggtggcg gcgggattgc cggccagggc ggccaggtgg ctgtagctga tgacctctcc
     4201 gaacttcacc actttcagca gtttccacag cacctggcgg gtaaagctct cctgctggaa
     4261 cactgggtgg tgcagggctg gcacagggaa ctcctcgatg gcctcaggct ggtgaaagta
     4321 ggcgttgagc caggcggtgg cctgcatcag tggctctggt ccgcccagca cggcggctgg
     4381 ggcaggcact tccacggcgt cggcggcaga tgttcctttg cccaggaaga tgatacggtg
     4441 caggccctgt tcgcacccag acagttccag cttgcccaga gggctatcca gggtggtgcg
     4501 cttcatttcg cagtctttgt cgatatctac cggtatactc cctgagccgc ctgatgcaga
     4561 gcccccaccg ctgccgctcc gaagaccgga gacgatcctc acggtgaacg tcttgggttg
     4621 cttcctcttg agagagctga agctcatgcg ggaagcgatg gccccggcca tggtggcact
     4681 agtaccggaa atagatcctc tagtagagtc ggtgtcttct atggaggtca aaacagcgtg
     4741 gatggcgtct ccaggcgatc tgacggttca ctaaacgagc tctgcttata tagacctccc
     4801 accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa
     4861 agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg
     4921 gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca
     4981 ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
     5041 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
     5101 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
     5161 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
     5221 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
     5281 taacgcggaa ctcccaagct tatcgataaa ctggatctct gctgtccctg taataaaccc
     5341 gaaaattttg aatttttgta atttgttttt gtaattcttt agtttgtatg tctgttgcta
     5401 ttatgtctac tattctttcc cctgcactgt accccccaat cccccctttt cttttaaagg
     5461 cgataccgtc gagatccgtt cactaatcga atggatctgt ctctgtctct ctctccacct
     5521 tcttcttcta ttccttcggg cctgtcgggt cccctcgggg ttgggaggtg ggtctgaaac
     5581 gataatggtg aatatccctg cctaactcta ttcactatag aaagtacagc aaaaactatt
     5641 cttaaaccta ccaagcctcc tactatcatt atgaataatt ttatatacca cagccaattt
     5701 gttatgttaa accaattcca caaacttgcc catttatcta attccaataa ttcttgttca
     5761 ttcttttctt gctggttttg cgattcttca attaaggagt gtattaagct tgtgtaattg
     5821 ttaatttctc tgtcccactc catccaggtc gtgtgattcc aaatctgttc cagagattta
     5881 ttactccaac tagcattcca aggcacagca gtggtgcaaa tgagttttcc agagcaaccc
     5941 caaatcccca ggagctgttg atcctttagg tatctttcca cagccaggat tcttgcctgg
     6001 agctgcttga tgccccagac tgtgagttgc aacagatgct gttgcgcctc aatagccctc
     6061 agcaaattgt tctgctgctg cactatacca gacaataatt gtctggcctg taccgtcagc
     6121 gtcattgacg ctgcgcccat agtgcttcct gctgctccca agaacccaag gaacaaagct
     6181 cctattccca ctgctctttt ttctctctgc accactcttc tctttgcctt ggtgggtgct
     6241 actcctaatg gttcaatttt tactacttta tatttatata attcacttct ccaattgtcc
     6301 ctcatatctc ctcctccagg tctgaagatc agcggccggc cgcttgctgt gcggtggtct
     6361 tacttttgtt ttgctcttcc tctatcttgt ctaaagcttc cttggtgtct tttatctcta
     6421 tcctttgatg cacacaatag agggttgcta ctgtattata taatgatcta agttcttctg
     6481 atcctgtctg aagggatggt tgtagctgtc ccagtatttg tctacagcct tctgatgttt
     6541 ctaacaggcc aggattaact gcgaatcgtt ctagctccct gcttgcccat actatatgtt
     6601 ttaatttata ttttttcttt ccccctggcc ttaaccgaat tttttcccat cgcgatctaa
     6661 ttctcccccg cttaatactg acgctctcgc acccatctct ctccttctag cctccgctag
     6721 tcaaaatttt tggcgtactc accagtcgcc gcccctcgcc tcttgccgtg cgcgcttcag
     6781 caagccgagt cctgcgtcga gagagctcct ctggtttccc tttcgctttc aagtccctgt
     6841 tcgggcgcca ctgctagaga ttttccacac tgactaaaag ggtctgaggg atctctagtt
     6901 accagagtca cacaacagac gggcacacac tacttgaagc actcaaggca agctttattg
     6961 aggcttaagc agtgggttcc ctagttagcc agagagctcc caggctcaga tctggtctaa
     7021 ccagagagac ccagtacagg caaaaagcag ctgcttatat gcaggatctg agggctcgcc
     7081 actccccagt cccgcccagg ccacgcctcc ctggaaagtc cccagcggaa agtcccttgt
     7141 agcaagctcg atatcagcag ttcttgaagt actccggatg cagctctcgg gccacgtgat
     7201 gaaatgctag gcggctgtca aacctccact ctaacacttc tctctccggg tcatccatcc
     7261 catgcaggct cacagggtgt aacaagctgg tgttctctcc tttattggcc tcttctacct
     7321 tatctggctc aactggtact agcttgtagc accatccaaa ggtcagtgga tatctgaccc
     7381 ctggccctgg tgtgtagttc tgctaatcag ggaagtagcc ttgtgtgtgg tagatccaca
     7441 gatcaaggat atcttgtctt ctttgggagt gaattagccc ttccaactac taagtttgta
     7501 gtacatattt aacaaataca atttctttaa aatgaaaata attcagagga atcacaggtt
     7561 tagagtaaat gaaaccacag gtaattggca gtggtaatag ggtatggggt gggaagtttg
     7621 ggatgatttt ggttagcttg agttatccag ttgatccaga catgataaga tacattgatg
     7681 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     7741 atgctattgc tttatttgta accattataa gctgcaataa acaagttgat ctcccgatcc
     7801 gtcgacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     7861 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     7921 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     7981 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     8041 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     8101 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     8161 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     8221 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     8281 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     8341 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     8401 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     8461 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     8521 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     8581 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     8641 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     8701 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     8761 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     8821 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     8881 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     8941 agaccccgta gaaaagatca aaggatcttc
//
