LOCUS Exported 9142 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Photo-activatable/light sensitive Vav2. ACCESSION . VERSION . KEYWORDS pBabe-Tet-off-puro-mCherry-PA(wt)-Vav2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9142) 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 9142) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9142 /organism="synthetic DNA construct" /mol_type="other DNA" 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 1399..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 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" 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 5556..5572 /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 5605..6104 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(5640..5663) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 6037..6056 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 6127..6726 /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(6127..6146) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 6623..6643 /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind complement(7235..7252) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7406..7994) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7486..7505) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8154..9014) /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 8777..8796 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9015..9119) /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 caatgcggcc gctggctcat cgactgcaag gtcctgccgc ccaaccaccg 2641 cgtcgtgtgg ccctcgggat caggtttggc tactacactt gaacgtattg agaagaactt 2701 tgtcattact gacccaagat tgccagataa tcccattata ttcgcgtccg atagtttctt 2761 gcagttgaca gaatatagcc gtgaagaaat tttgggaaga aactgcaggt ttctacaagg 2821 tcctgaaact gatcgcgcga cagtgagaaa aattagagat gccatagata accaaacaga 2881 ggtcactgtt cagctgatta attatacaaa gagtggtaaa aagttctgga acctctttca 2941 cttgcagcct atgcgagatc agaagggaga tgtccagtac tttattgggg ttcagttgga 3001 tggaactgag catgtccgag atgctgccga gagagaggga gtcatgctga ttaagaaaac 3061 tgcagaaaat attgatgagg cggcaaaaga acttggcagt ggtgcggtgg tcttcgacct 3121 ggcgcaggcg ctgcgcgacg gcgtccttct gtgccagctg ctgcacaacc tctcccccgg 3181 ttccatcgac cttaaggaca tcaacttccg gccgcagatg tcccagttct tgtgtctgaa 3241 gaatattcgc accttcctca aagtctgcca tgacaaattt ggactaagaa acagtgagct 3301 gtttgaccct ttcgacctct ttgatgtccg agacttcggg aaggtcatct ctgcagtgtc 3361 ccgtctgtcc ctgcacagca tcgcacagag caaagggatc aggccatttc catcagagga 3421 gacagctgaa aatgatgacg acgtctaccg gagcctggaa gagctggctg acgagcatga 3481 cctgggtgag gacatctacg actgtgtccc gtgtgaagat gaaggagatg acatttatga 3541 ggacatcatc aaggtggagg tgcagcagcc catgaaaatg ggcatgactg aggacgacaa 3601 gagaagctgc tgcttgttag agattcagga gaccgaggcc aagtactacc gcaccctgga 3661 ggacattgag aagaactaca tgggtccctt gcggctggtg ctgagcccgg cggatatggc 3721 tgctgtcttc atcaacctgg aggacctcat caaggtgcat cacagctttc tgcgagccat 3781 cgatgtgtcc atgatggctg gtggcagtac cctggctaag gtctttctgg agtttaagga 3841 aaggctcctg atctatggag agtactgtag ccacatggaa cacgctcaga gtacactgaa 3901 ccagctcctc gccagccgag aggacttcag gcagaaagtg gaggagtgca cactcagggt 3961 tcaggatggc aagttcaagc tgcaagacct gctggtggtg cccatgcaac gggtgctgaa 4021 gtaccacctg ctgctcaagg agctcctgag ccattctgca gaccgaccag aaagacaaca 4081 gctcaaagaa gccctggaag ccatgcagga cttggccatg tacatcaatg aagtgaagcg 4141 ggacaaggag accttgaaga agattagcga gttccagtgc tccatagaaa acctgcaagt 4201 gaagctggag gaatttggga ggccaaagat tgacggggag cttaaagtcc ggtccatagt 4261 caaccacacc aagcaagaca ggtacctgtt cctatttgac aaggtggtca tcgtgtgcaa 4321 gaggaagggc tacagctatg agctgaagga ggtcattgag ctgctcttcc acaagatgac 4381 cgatgacccg atgcacaaca aggacatcaa gaagtggtcc tatggcttct acctgattca 4441 cctccaagga aagcaaggct ttcagttctt ctgcaagacg gaagacatga agcggaagtg 4501 gatggagcag ttcgagatgg ccatgtcaaa catcaagcca gataaggcca atgccaacca 4561 tcatagcttc cagatgtaca cattcgacaa gactaccaac tgcaaagcct gcaagatgtt 4621 tctcaggggt accttctacc agggatacct gtgtaccaga tgtggcgtcg gggcacacaa 4681 ggaatgcctg gaggtgatcc ccccctgcaa gatgagttca cctgcagatg tggatgctcc 4741 tggagcagga ccaggtccca agatggttgc tgtgcagaat taccatggta acccagcccc 4801 tcctgggaag cctgtgttga ccttccagac aggagacgtg atagaactgc tccggggtga 4861 ccccgattct ccttggtggg aagggcggct ggttcaaacc aggaagtcag ggtattttcc 4921 cagctcatct gtgaagccct gcccagtgga tggaaggccg cctactggcc gcccaccatc 4981 ccgggagatc gattacactg cctacccgtg gtttgctggc aacatggaac ggcagcagac 5041 ggacaatctg ctcaaatctc atgccagtgg gacctacctc atcagagagc gtccagcaga 5101 ggcagaacgc ttcgccatca gcatcaagtt caatgatgag gtgaaacata tcaaggtggt 5161 ggagaaagat agctggatcc acatcacgga agctaagaag tttgagagcc tcctggagct 5221 ggtggagtac taccagtgtc actcactcaa ggaaagcttc aagcagttgg acactacact 5281 caagttcccc tacaagtctc gggagcgaac caccagcagg gcctcaagcc ggtctccagt 5341 gttcacaccg cgggtcatcg gcacagccgt ggccaggtac aactttgctg ctcgagacat 5401 gcgggagctg tcactgcgag aaggtgatgt ggtgaagatc tacagccgaa ttggtgggga 5461 ccagggctgg tggaaaggag agacaaatgg gcggatcggt tggtttcctt cgacgtacgt 5521 ggaagaggaa ggcgtccagt gagtcgagtg tggaattgtg agcggataac aatttgtcga 5581 gggcccctgc aggtcaattc taccgggtag gggaggcgct tttcccaagg cagtctggag 5641 catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca 5701 cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc 5761 caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca 5821 ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg 5881 agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt 5941 tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg 6001 gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca 6061 cgtctgccgc gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agccaataag 6121 cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg 6181 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat 6241 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg 6301 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg 6361 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg 6421 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc 6481 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt 6541 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc 6601 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc 6661 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt 6721 gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca 6781 tcgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc 6841 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa 6901 ctgagaatag agaagttcag atcaaggtag gagatccctg agcccacaac ccctcactcg 6961 gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct 7021 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg 7081 actacccgtc agcgggggtc tttcacatgc agcatgtatc aaaattaatt tggttttttt 7141 tcttaagtat ttacattaaa tggccatagt tgcattaatg aatcggccaa cgcgcgggga 7201 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 7261 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 7321 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 7381 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 7441 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 7501 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 7561 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 7621 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 7681 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 7741 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 7801 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 7861 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 7921 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 7981 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 8041 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 8101 ttttgcggcc gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 8161 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 8221 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 8281 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 8341 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 8401 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 8461 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 8521 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 8581 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 8641 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 8701 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 8761 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 8821 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 8881 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 8941 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 9001 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 9061 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 9121 acatttcccc gaaaagtgcc ac //