LOCUS       Exported                8018 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synchronize trafficking of VSVG from the ER (RUSH system).
ACCESSION   .
VERSION     .
KEYWORDS    Str-Ii_VSVG-SBP-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8018)
  AUTHORS   Boncompain G, Divoux S, Gareil N, de Forges H, Lescure A, Latreche 
            L, Mercanti V, Jollivet F, Raposo G, Perez F
  TITLE     Synchronization of secretory protein traffic in populations of 
            cells.
  JOURNAL   Nat Methods. 2012 Mar 11;9(5):493-8. doi: 10.1038/nmeth.1928.
  PUBMED    22406856
REFERENCE   2  (bases 1 to 8018)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8018
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(48..67)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        239..618
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..822
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     773..793
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     867..886
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        867..885
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1459..1485
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          2220..2449
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2432..2451
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2522..3092
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2686..2703)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2913..2932
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3119..4651
                     /codon_start=1
                     /product="vesicular stomatitis virus G glycoprotein"
                     /label=VSV-G
                     /note="Indiana strain"
                     /translation="MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSD
                     LNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPS
                     VEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQ
                     FINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFR
                     SNYFAYETGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQT
                     SVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLK
                     YFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLRTSSGYKFP
                     LYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWF
                     SSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK"
     CDS             4661..4774
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     CDS             4784..5488
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(4926..4944)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5194..5213
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5642..5763
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5679..5698)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5865..5881)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5865..5881)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5878..5900)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5889..5905
                     /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        complement(5913..5943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5958..5979
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6096..6113)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6267..6855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6347..6366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7026..7886)
                     /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     7649..7668
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7887..7991)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
       61 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
      121 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
      181 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
      241 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      301 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      361 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      421 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      481 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      541 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      601 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      661 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      721 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      781 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa
      841 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagcttgg
      901 taccgagctc ggatcgatat catgcaccgg aggagatcac gctcttgtag ggaggaccag
      961 aaacctgtca ccggtgaccc tagcaaagac tcaaaagctc aggtgtccgc tgccgaggct
     1021 ggcattactg gaacatggta caatcagctc gggagcacct ttattgtgac tgctggagcc
     1081 gatggagccc tcaccggaac atacgaatct gctgtgggaa acgccgaatc acggtacgtc
     1141 ctcactggcc gatacgatag tgcccctgcc accgacggat ctgggactgc cctgggatgg
     1201 actgtcgctt ggaaaaacaa ctaccggaat gctcattctg ccacaacatg gagtggacag
     1261 tacgtgggag gcgctgaggc tagaatcaat acacagtggc tgctcacatc tggcacaacc
     1321 gaggcaaatg cttggaaatc caccctggtg ggacatgaca cattcaccaa agtgaaaccc
     1381 tccgccgctt caatcgatgc cgccaaaaaa gccggagtca acaacggcaa tcctctggat
     1441 gccgtccagc aggtcgacta tccgtacgac gtaccagact acgcagtcgg accgatggac
     1501 gatcagaggg acctcattag caacaacgaa cagctgccta tgctgggacg gcgacctgga
     1561 gcccctgaat ccaaatgctc taggggagca ctgtacactg gcttctccat tctcgtgaca
     1621 ctgctgctgg ccgggcaggc tactactgct tacttcctgt accagcagca ggggcggctg
     1681 gacaaactca ctgtgacatc tcagaacctc cagctggaaa atctgaggat gaaactgccc
     1741 aaacccccta aacccgtgtc caaaatgagg atggccacac ctctgctcat gcaggcactg
     1801 ccaatgggag ccctgcccca ggggcccatg cagaatgcca ccaagtatgg caacatgaca
     1861 gaggaccatg tgatgcacct gctccagaat gctgaccccc tgaaggtgta cccgccactg
     1921 aaggggagct tcccggagaa cctgagacac cttaagaaca ccatggagac catagactgg
     1981 aaggtctttg agagctggat gcaccattgg ctcctgtttg aaatgagcag gcactccttg
     2041 gagcaaaagc ccactgacgc tccaccgaaa gagtcactgg aactggagga cccgtcttct
     2101 gggctgggtg tgaccaagca ggatctgggc ccagtcccca tgtgaggatc cgcggccgca
     2161 tagataactg atccagtgtg ctggaattaa ttcgctgtct gcgagggcca gctgttgggg
     2221 tgagtactcc ctctcaaaag cgggcatgac ttctgcgcta agattgtcag tttccaaaaa
     2281 cgaggaggat ttgatattca cctggcccgc ggtgatgcct ttgagggtgg ccgcgtccat
     2341 ctggtcagaa aagacaatct ttttgttgtc aagcttgagg tgtggcaggc ttgagatctg
     2401 gccatacact tgagtgacaa tgacatccac tttgcctttc tctccacagg tgtccactcc
     2461 caggtccaac tgcaggtcga gcatgcatct agggcggcca attccgcccc tctccccccc
     2521 ccccctctcc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     2581 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     2641 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     2701 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     2761 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     2821 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     2881 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     2941 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca
     3001 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     3061 acgtggtttt cctttgaaaa acacgatgat aagcttgcca caacccggga ggcgcgccat
     3121 gaagtgcctt ttgtacttag cctttttatt cattggggtg aattgcaagt tcaccatagt
     3181 ttttccacac aaccaaaaag gaaactggaa aaatgttcct tctaattacc attattgccc
     3241 gtcaagctca gatttaaatt ggcataatga cttaataggc acagccttac aagtcaaaat
     3301 gcccaagagt cacaaggcta ttcaagcaga cggttggatg tgtcatgctt ccaaatgggt
     3361 cactacttgt gatttccgct ggtatggacc gaagtatata acacattcca tccgatcctt
     3421 cactccatct gtagaacaat gcaaggaaag cattgaacaa acgaaacaag gaacttggct
     3481 gaatccaggc ttccctcctc aaagttgtgg atatgcaact gtgacggatg ccgaagcagt
     3541 gattgtccag gtgactcctc accatgtgct ggttgatgaa tacacaggag aatgggttga
     3601 ttcacagttc atcaacggaa aatgcagcaa ttacatatgc cccactgtcc ataactctac
     3661 aacctggcat tctgactata aggtcaaagg gctatgtgat tctaacctca tttccatgga
     3721 catcaccttc ttctcagagg acggagagct atcatccctg ggaaaggagg gcacagggtt
     3781 cagaagtaac tactttgctt atgaaactgg aggcaaggcc tgcaaaatgc aatactgcaa
     3841 gcattgggga gtcagactcc catcaggtgt ctggttcgag atggctgata aggatctctt
     3901 tgctgcagcc agattccctg aatgcccaga agggtcaagt atctctgctc catctcagac
     3961 ctcagtggat gtaagtctaa ttcaggacgt tgagaggatc ttggattatt ccctctgcca
     4021 agaaacctgg agcaaaatca gagcgggtct tccaatctct ccagtggatc tcagctatct
     4081 tgctcctaaa aacccaggaa ccggtcctgc tttcaccata atcaatggta ccctaaaata
     4141 ctttgagacc agatacatca gagtcgatat tgctgctcca atcctctcaa gaatggtcgg
     4201 aatgatcagt ggaactacca cagaaaggga actgtgggat gactgggcac catatgaaga
     4261 cgtggaaatt ggacccaatg gagttctgag gaccagttca ggatataagt ttcctttata
     4321 catgattgga catggtatgt tggactccga tcttcatctt agctcaaagg ctcaggtgtt
     4381 cgaacatcct cacattcaag acgctgcttc gcaacttcct gatgatgaga gtttattttt
     4441 tggtgatact gggctatcca aaaatccaat cgagcttgta gaaggttggt tcagtagttg
     4501 gaaaagctct attgcctctt ttttctttat catagggtta atcattggac tattcttggt
     4561 tctccgagtt ggtatccatc tttgcattaa attaaagcac accaagaaaa gacagattta
     4621 tacagacata gagatgaacc gacttggaaa gctgaattcc atggacgaga agaccactgg
     4681 ttggcgaggt ggacacgttg ttgaaggact ggctggggaa cttgaacaac ttcgtgcacg
     4741 actggagcat cacccacaag gtcaacgtga accacctgca ggtgtgagca agggcgagga
     4801 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
     4861 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     4921 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
     4981 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
     5041 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     5101 cggcgtggtg accgtgaccc aggactcctc cctacaggac ggcgagttca tctacaaggt
     5161 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
     5221 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     5281 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
     5341 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
     5401 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
     5461 caccggcggc atggacgagc tgtacaaggg ccggccttaa ggcctcgagg gccaattaat
     5521 taactctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     5581 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     5641 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5701 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5761 ttaacgcgtc gagtgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5821 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5881 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5941 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6001 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6061 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6121 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6181 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6241 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6301 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6361 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6421 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6481 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6541 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6601 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6661 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6721 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6781 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6841 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6901 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6961 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7021 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7081 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7141 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7201 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7261 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7321 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7381 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7441 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7501 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7561 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7621 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7681 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7741 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7801 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7861 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7921 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7981 ggggttccgc gcacatttcc ccgaaaagtg ccacctga
//
