LOCUS       sequence-191108-        8503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rapamycin-activatable Vav2;DPZ: DH-PH-ZnF domains of Vav2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8503)
  AUTHORS   Dagliyan O, Tarnawski M, Chu PH, Shirvanyants D, Schlichting I, 
            Dokholyan NV, Hahn KM
  TITLE     Engineering extrinsic disorder to control protein activity in living
            cells.
  JOURNAL   Science. 2016 Dec 16;354(6318):1441-1444.
  PUBMED    27980211
REFERENCE   2  (bases 1 to 8503)
  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: "Science."; 
            date: "2016-12-16"; volume: "354(6318)"; pages: "1441-1444"
COMMENT     Sequence Label: pBabe-Tet-off-uniRapR-mCherry-Vav2(DPZ)
FEATURES             Location/Qualifiers
     source          1..8503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             942..1358
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1288..1310
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1326..1342
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    join(1399..1417,1418..1440,1441..1459,1460..1482,1483..1501,
                     1502..1524,1525..1543,1544..1566,1567..1585,1586..1608,
                     1609..1627,1628..1650,1651..1669)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        1702..1740
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1737..1761
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1856..1865
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1862..2569
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2007..2025)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2275..2294
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    4917..4933
                     /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)."
     promoter        4966..5465
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5001..5024)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5398..5417
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             5488..6087
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5488..5507)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5984..6004
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6596..6613)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6767..7355)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6847..6866)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7515..8306,8307..8375))
                     /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     8138..8157
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8376..8480)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
      901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
      961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatccac gaggcccttt
     1381 cgtcgtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac
     1441 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
     1501 aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt
     1561 taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg
     1621 atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
     1681 cgagctcggt acccgggtcg aggtaggcgt gtacggtggg aggcctatat aagcagagct
     1741 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
     1801 agacaccggg accgatccag cctccgcggc cccagatccg taaagtgaat tcgccgccac
     1861 catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     1921 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     1981 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     2041 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     2101 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     2161 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     2221 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     2281 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     2341 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     2401 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     2461 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     2521 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt ccggaatgga
     2581 gcagtggcgg caatgcggac gctggctcat cgactgcaag gtcctgccgc ccaaccaccg
     2641 cgtcgtgtgg ccctcggcgg tggtcttcga cctggcgcag gcgctgcgcg acggcgtcct
     2701 tctgtgccag ctgctgcaca acctctcccc cggttccatc gaccttaagg acatcaactt
     2761 ccggccgcag atgtcccagt tcttgtgtct gaagaatatt cgcaccttcc tcaaagtctg
     2821 ccatgacaaa tttggactaa gaaacagtga gctgtttgac cctttcgacc tctttgatgt
     2881 ccgagacttc gggaaggtca tctctgcagt gtcccgtctg tccctgcaca gcatcgcaca
     2941 gagcaaaggg atcaggccat ttccatcaga ggagacagct gaaaatgatg acgacgtcga
     3001 acggagcctg gaagagctgg ctgacgagca tgacctgggt gaggacatcg aagactgtgt
     3061 cccgtgtgaa gatgaaggag atgacattga agaggacatc atcaaggtgg aggtgcagca
     3121 gcccatgaaa atgggaatga cagaagacga caagagaagc tgctgcttgt tagagattca
     3181 ggagaccgag gccaagtact accgcaccct ggaggacatt gagaagaact acatgggtcc
     3241 cttgcggctg gtgctgagcc cggcggatat ggctgctgtc ttcatcaacc tggaggacct
     3301 catcaaggtg catcacagct ttctgcgagc catcgatgtg tccatgatgg ctggtggcag
     3361 taccctggct aaggtctttc tggagtttaa ggaaaggctc ctgatctatg gagagtactg
     3421 tagccacatg gaacacgctc agagtacact gaaccagctc ctcgccagcc gagaggactt
     3481 caggcagaaa gtggaggagt gcacactcag ggttcaggat ggcaagttca agctgcaaga
     3541 cctgctggtg gtgcccatgc aacgggtgct gaagtaccac ctgctgctca aggagctcct
     3601 gagccattct gcaccaacct gcgtggtgca ctacaccggg atgcttgaag atggaaagaa
     3661 atttgattcc tcccgggaca gaaacaagcc ctttaagttt atgctaggca agcaggaggt
     3721 gatccgaggc tgggaagaag gggttgccca gatgagtgtg ggtcagagag ccaaactgac
     3781 tatatctcca gattatgcct atggtgccac tgggcacggt tcgggctccg gatcaggcgt
     3841 caaggacctc ctccaagcct gggacctcta ttatcatgtg ttccgacgaa tctcaggtcc
     3901 tccaggacct ggatcaggtc tctggcatga gatgtggcat gaaggcctgg aagaggcatc
     3961 tcgtttgtac tttggggaaa ggaacgtgaa aggcatgttt gaggtgctgg agcccttgca
     4021 tgctatgatg gaacggggcc cccagactct gaaggaaaca tcctttaatc aggcctatgg
     4081 tcgagattta atggaggccc aagagtggtg caggaagtac atgaaatcag ggtcatcagg
     4141 gggctccgga tcaggcatca tcccaccaca tgccactctc gtcttcgatg tggagcttct
     4201 aaaactggaa cccgaccgac cagaaagaca acagctcaaa gaagccctgg aagccatgca
     4261 ggacttggcc atgtacatca atgaagtgaa gcgggacaag gagaccttga agaagattag
     4321 cgagttccag tgctccatag aaaacctgca agtgaagctg gaggaatttg ggaggccaaa
     4381 gattgacggg gagcttaaag tccggtccat agtcaaccac accaagcaag acaggtacct
     4441 gttcctattt gacaaggtgg tcatcgtgtg caagaggaag ggctacagct atgagctgaa
     4501 ggaggtcatt gagctgctct tccacaagat gaccgatgac ccgatgcaca acaaggacat
     4561 caagaagtgg tcctatggct tctacctgat tcacctccaa ggaaagcaag gctttcagtt
     4621 cttctgcaag acggaagaca tgaagcggaa gtggatggag cagttcgaga tggccatgtc
     4681 aaacatcaag ccagataagg ccaatgccaa ccatcatagc ttccagatgt acacattcga
     4741 caagactacc aactgcaaag cctgcaagat gtttctcagg ggtaccttct accagggata
     4801 cctgtgtacc agatgtggcg tcggggcaca caaggaatgc ctggaggtga tccccccctg
     4861 caagatgagt tcacctgcag atgtggatgc tcctggagca taactcgagt gtggaattgt
     4921 gagcggataa caatttgtcg agggcccctg caggtcaatt ctaccgggta ggggaggcgc
     4981 ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca
     5041 caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt
     5101 tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg
     5161 ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg
     5221 tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat
     5281 agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg
     5341 ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat
     5401 tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc
     5461 tttcgacctg cagccaataa gcttaccatg accgagtaca agcccacggt gcgcctcgcc
     5521 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     5581 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     5641 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     5701 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     5761 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     5821 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     5881 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     5941 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     6001 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     6061 tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa
     6121 aggagcgcac gaccccatgc atcgataaaa taaaagattt tatttagtct ccagaaaaag
     6181 gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc
     6241 aaggcatgga aaaatacata actgagaata gagaagttca gatcaaggta ggagatccct
     6301 gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc
     6361 cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg
     6421 agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcacatg cagcatgtat
     6481 caaaattaat ttggtttttt ttcttaagta tttacattaa atggccatag ttgcattaat
     6541 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6601 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6661 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6721 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6781 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6841 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6901 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6961 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7021 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7081 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7141 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7201 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7261 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7321 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7381 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7441 aaaggatctt cacctagatc cttttgcggc cgcaaatcaa tctaaagtat atatgagtaa
     7501 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7561 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7621 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7681 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7741 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7801 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7861 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7921 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7981 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     8041 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     8101 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     8161 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     8221 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     8281 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     8341 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     8401 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     8461 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac
//
