LOCUS       Exported                8679 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  PI(WT): Photo-inhibitable/blue light sensitive; kinase dead Src 
            mutation.
ACCESSION   .
VERSION     .
KEYWORDS    pBabe-PI(WT)-Src(KD)-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8679)
  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 8679)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8679
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     131..150
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             285..1310
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     complement(1787..1804)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1958..2546)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2038..2057)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2706..3566)
                     /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     3329..3348
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3567..3671)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             3702..4171
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    4236..4435
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             4636..5052
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4982..5004
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5020..5036
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    5093..5363
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        5396..5434
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5431..5455
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      5547..5556
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      7599..7608
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             7605..8315
                     /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(7750..7768)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     8018..8037
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    8329..8345
                     /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        8378..198
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8413..8436)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt
       61 gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc
      121 ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc
      181 tcatctccgg gcctttcgac ctgcagccaa taagcttgcc agctggggcg ccctctggta
      241 aggttgggaa gccctgcaga tctgatcccg gggggcaatg agatatgaaa aagcctgaac
      301 tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga
      361 tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat
      421 atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc
      481 actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa tttagcgaga
      541 gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa
      601 ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg
      661 atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta
      721 catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga
      781 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg
      841 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga
      901 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc
      961 aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga
     1021 cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata
     1081 tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg
     1141 cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc
     1201 gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg
     1261 ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag agtagatgcc
     1321 gaccgggatc tatcgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg
     1381 aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg
     1441 aaaaatacat aactgagaat agagaagttc agatcaaggt aggagatccc tgagcccaca
     1501 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
     1561 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     1621 tctgagtgat tgactacccg tcagcggggg tctttcacat gcagcatgta tcaaaattaa
     1681 tttggttttt tttcttaagt atttacatta aatggccata gttgcattaa tgaatcggcc
     1741 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     1801 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     1861 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     1921 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1981 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2041 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2101 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2161 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2221 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2281 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2341 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     2401 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2461 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     2521 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2581 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2641 tcacctagat ccttttgcgg ccgcaaatca atctaaagta tatatgagta aacttggtct
     2701 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2761 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     2821 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     2881 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     2941 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3001 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3061 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3121 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3181 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3241 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3301 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3361 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3421 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3481 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3541 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     3601 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     3661 ggggttccgc gcacatttcc ccgaaaagtg ccacctgcag cctgaatatg ggccaaacag
     3721 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga
     3781 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     3841 agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc
     3901 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     3961 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     4021 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
     4081 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     4141 tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga tcgggagacc
     4201 cctgcccagg gaccaccgac ccaccaccgg gaggcaagct ggccagcaac ttatctgtgt
     4261 ctgtccgatt gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta ctagttagct
     4321 aactagctct gtatctggcg gacccgtggt ggaactgacg agttctgaac acccggccgc
     4381 aaccctggga gacgtcccag ggactttggg ggccgttttt gtggcccgac ctgaggaagg
     4441 gagtcgatgt ggaatccgac cccgtcagga tatgtggttc tggtaggaga cgagaaccta
     4501 aaacagttcc cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct
     4561 tgtctgctgc agcgctgcag catcgttctg tgttgtctct gtctgactgt gtttctgtat
     4621 ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt aggtcactgg
     4681 aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt
     4741 accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt
     4801 aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac
     4861 ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg
     4921 gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc
     4981 ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct
     5041 tctctaggcg ccggccggat ccacgaggcc ctttcgtcgt cgagtttacc actccctatc
     5101 agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga
     5161 aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc
     5221 cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa
     5281 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta
     5341 ccactcccta tcagtgatag agaaaagtga aagtcgagct cggtacccgg gtcgaggtag
     5401 gcgtgtacgg tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgcctg
     5461 gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg
     5521 cggccccaga tccgtaaagt gaattcgcca ccatgggcag caacaagagc aagcccaagg
     5581 acgccagcca gcggcgccgc agcctggagc cctcggaaaa cgtgcacggg gcagggggcg
     5641 ccttcccggc ctcacagaca ccgagcaagc ccgcctccgc cgacggccac cgcgggccca
     5701 gcgccgcctt cgtgccgccc gcggccgagc ccaagctctt cggaggcttc aactcctcgg
     5761 acaccgtcac ctccccgcag agggcggggc ctctggcagg tggggtgacc acctttgtgg
     5821 ccctctatga ctatgagtca cggacagaga ctgacctgtc cttcaagaaa ggggagcggc
     5881 tgcagattgt taacaacacg gagggagact ggtggctggc acactcgctg agcacgggac
     5941 agaccggtta catccccagc aactatgtgg cgccctccga ctccatccag gctgaggagt
     6001 ggtactttgg caagatcact agacgggaat cagagcggct gctgctcaac gccgagaacc
     6061 cgagagggac cttcctcgtg agggagagtg agaccacaaa aggtgcctac tgcctctctg
     6121 tatccgactt cgacaatgcc aagggcctaa atgtgaaaca ctacaagatc cgcaagctgg
     6181 acagcggcgg tttctacatc acctcccgca cccagttcaa cagcctgcag cagctcgtgg
     6241 cttactactc caaacatgct gatggcctgt gtcaccgcct cactaccgta tgtcccacat
     6301 ccaagcctca gacccaggga ttggccaagg atgcgtggga gatcccccgg gagtccctgc
     6361 ggctggaggt caagctgggc cagggttgct tcggagaggt gtggatgggg acctggaact
     6421 tggctactac acttgaacgt attgagaaga actttgtcat tactgaccca agattgccag
     6481 ataatcccat tatattcgcg tccgatagtt tcttgcagtt gacagaatat agccgtgaag
     6541 aaattttggg aagaaactgc aggtttctac aaggtcctga aactgatcgc gcgacagtga
     6601 gaaaaattag agatgccata gataaccaaa cagaggtcac tgttcagctg attaattata
     6661 caaagagtgg taaaaagttc tggaacctct ttcacttgca gcctatgcga gatcagaagg
     6721 gagatgtcca gtactttatt ggggttcagt tggatggaac tgagcatgtc cgagatgctg
     6781 ccgagagaga gggagtcatg ctgattaaga aaactgcaga aaatattgat gaggcggcaa
     6841 aagaacttac cacgagggtt gccatcaaaa ctctgaagcc aggcaccatg tccccagagg
     6901 ccttcctgca ggaggcccaa gtcatgaaga aactgaggca cgagaaactg gtgcagctgt
     6961 atgctgtggt gtcggaagaa cccatttaca ttgtgacaga gtacatgaac aaggggagtc
     7021 tgctggactt tctcaagggg gaaacgggca aatatttgcg gctaccccag ctggtggaca
     7081 tgtctgctca gatcgcttca ggcatggcct atgtggagcg gatgaactat gtgcaccggc
     7141 gccttcgagc cgccaatatc ctagtagggg agaacctggt gtgcaaagtg gccgactttg
     7201 ggttggcccg gctcatagaa gacaacgaat acacagcccg gcaaggtgcc aaattcccca
     7261 tcaagtggac cgcccctgaa gctgctctgt acggcaggtt caccatcaag tcggatgtgt
     7321 ggtcctttgg gattctgctg accgagctca ccactaaggg aagagtgccc tatcctggga
     7381 tggtgaaccg tgaggttctg gaccaggtgg agcggggcta ccggatgcct tgtccccccg
     7441 agtgccccga gtccctgcat gaccttatgt gccagtgctg gcggaaggag cccgaggagc
     7501 ggcccacctt cgagtacctg caggccttcc tggaagacta ctttacgtcc actgagccac
     7561 agtaccagcc cggggagaac ctatcggatc caccggtcgc caccatggtg agcaagggcg
     7621 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
     7681 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     7741 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
     7801 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     7861 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     7921 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
     7981 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
     8041 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
     8101 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
     8161 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
     8221 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     8281 actccaccgg cggcatggac gagctgtaca agtaagtcga gtgtggaatt gtgagcggat
     8341 aacaatttgt cgagggcccc tgcaggtcaa ttctaccggg taggggaggc gcttttccca
     8401 aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc
     8461 ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt
     8521 ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag
     8581 ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg
     8641 gacagcaccg ctgagcaatg gaagcgggta ggcctttgg
//
