LOCUS       Exported                6277 bp ds-DNA     circular SYN 20-APR-2021
DEFINITION  synchronize trafficking of Ecadherin from the ER (RUSH system).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6277)
  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 6277)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Apr 21, 2021 from SnapGene 5.2.4
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2012-03-11- 11"; volume: "9"; 
            issue: "5"; pages: "493-8"
FEATURES             Location/Qualifiers
     source          1..6277
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             954..1577
                     /codon_start=1
                     /label=Str-KDEL
                     /translation="MDVCVRLALWLLWGLLLHQGQSLSHSHSEKATGTSSGAGLNDPSK
                     DSKAQVSAAEAGITGTWYNQLGSTFIVTAGADGALTGTYESAVGNAESRYVLTGRYDSA
                     PATDGSGTALGWTVAWKNNYRNAHSATTWSGQYVGGAEARINTQWLLTSGTTEANAWKS
                     TLVGHDTFTKVKPSAASIDAAKKAGVNNGNPLDAVQQLGPKDEL"
     intron          1646..1875
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1858..1877
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    1933..2506
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2100..2117)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2327..2346
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2533..3765
                     /codon_start=1
                     /label=EPV-CXCL9-mCherry-SBP
                     /translation="MKKSGVLFLLGIILLVLIGVQGEPVVRKGRCSCISTNQGTIHLQS
                     LKDLKQFAPSPSCEKIEIIATLKNGVQTCLNPDSADVKELIKKWEKQVSQKKKQKNGKK
                     HQKKKVLKVRKSQRSRQKKTTVDSGSSGSMVSKGEEDNMAIIKEFMRFKVHMEGSVNGH
                     EFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLK
                     LSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGW
                     EASSERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDIT
                     SHNEDYTIVEQYERAEGRHSTGGMDELYKGSGSDEKTTGWRGGHVVEGLAGELEQLRAR
                     LEHHPQGQREP"
     CDS             2533..2907
                     /codon_start=1
                     /label=CXCL9 (T23E)
                     /translation="MKKSGVLFLLGIILLVLIGVQGEPVVRKGRCSCISTNQGTIHLQS
                     LKDLKQFAPSPSCEKIEIIATLKNGVQTCLNPDSADVKELIKKWEKQVSQKKKQKNGKK
                     HQKKKVLKVRKSQRSRQKKTT"
     CDS             2932..3639
                     /codon_start=1
                     /label=mCherry
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             3652..3765
                     /codon_start=1
                     /label=SBP
                     /translation="DEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     polyA_signal    3897..4018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3934..3953)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4120..4136)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4120..4136)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4133..4155)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4144..4160
                     /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(4168..4198)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4213..4234
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4351..4368)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4522..5110)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4602..4621)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5281..6141)
                     /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     5904..5923
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6142..6246)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgatatctgc ggcctagcta gcgcttaagg cctgttaacc ggtatggatg
      961 tatgcgtccg tcttgccctg tggctcctct ggggactcct cctgcaccag ggccagagcc
     1021 tcagccatag tcacagtgag aaggcgacag gaaccagctc gggcgccggt ttaaacgacc
     1081 ctagcaaaga ctcaaaagct caggtgtccg ctgccgaggc tggcattact ggaacatggt
     1141 acaatcagct cgggagcacc tttattgtga ctgctggagc cgatggagcc ctcaccggaa
     1201 catacgaatc tgctgtggga aacgccgaat cacggtacgt cctcactggc cgatacgata
     1261 gtgcccctgc caccgacgga tctgggactg ccctgggatg gactgtcgct tggaaaaaca
     1321 actaccggaa tgctcattct gccacaacat ggagtggaca gtacgtggga ggcgctgagg
     1381 ctagaatcaa tacacagtgg ctgctcacat ctggcacaac cgaggcaaat gcttggaaat
     1441 ccaccctggt gggacatgac acattcacca aagtgaaacc ctccgccgct tcaatcgatg
     1501 ccgccaaaaa agccggagtc aacaacggca atcctctgga tgccgtccag cagctcggac
     1561 cgaaagatga actgtgagcg gccgcataga taactgatcc agtgtgctgg aattaattcg
     1621 ctgtctgcga gggccagctg ttggggtgag tactccctct caaaagcggg catgacttct
     1681 gcgctaagat tgtcagtttc caaaaacgag gaggatttga tattcacctg gcccgcggtg
     1741 atgcctttga gggtggccgc gtccatctgg tcagaaaaga caatcttttt gttgtcaagc
     1801 ttgaggtgtg gcaggcttga gatctggcca tacacttgag tgacaatgac atccactttg
     1861 cctttctctc cacaggtgtc cactcccagg tccaactgca ggtcgagcat gcatctaggg
     1921 cggccaattc cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga
     1981 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     2041 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     2101 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     2161 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     2221 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     2281 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     2341 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     2401 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     2461 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagctt gccacaaccc
     2521 gggaggcgcg ccatgaagaa gtctggagtc ctgttcctct tggggatcat tctgttggta
     2581 ctgataggtg tacaaggtga accagtcgta agaaaagggc ggtgttcctg catttccacc
     2641 aatcaaggga cgatccatct ccaatccctt aaagacctga agcaatttgc tccaagcccc
     2701 tcttgtgaga agattgaaat catcgctaca ctgaagaatg gagtacaaac atgcctgaac
     2761 ccggattctg cagacgtcaa agagctcatc aagaaatggg agaaacaagt ttctcaaaag
     2821 aagaaacaaa agaacggcaa gaagcatcag aagaagaaag tactgaaggt gcggaaaagc
     2881 caaaggtctc gccaaaagaa gacaaccgtc gactcaggca gtagtggaag catggtcagc
     2941 aagggagaag aggacaacat ggctatcata aaagagttca tgaggtttaa agtccatatg
     3001 gaaggcagcg tcaatggtca cgagttcgag atagaaggag agggcgaggg aaggccatac
     3061 gaaggtacgc agacagccaa gctgaaagtg accaaaggcg gtccccttcc cttcgcatgg
     3121 gacattctta gtccccagtt tatgtacggg tctaaagcgt atgttaaaca ccccgcggac
     3181 attccggatt acctgaagct gtcttttccg gaaggattta agtgggagcg ggtcatgaat
     3241 tttgaagacg gtggggtcgt gaccgttacc caagattcca gcctgcaaga cggagaattt
     3301 atctacaagg tcaagctcag gggcacaaat ttcccatccg atggacctgt gatgcaaaag
     3361 aaaactatgg gatgggaagc ctcaagcgag agaatgtatc cagaggacgg tgcattgaaa
     3421 ggagaaatta agcaaagact caaactcaaa gatggcgggc attacgacgc tgaggtcaag
     3481 accacctata aggcgaagaa acccgtccag ctgcccggcg cctataatgt taacattaaa
     3541 ctggacatca caagccataa cgaagattat acaatcgtcg agcagtacga gcgagcggaa
     3601 ggacggcata gcactggcgg gatggatgag ctttacaagg gatccggaag cgacgagaag
     3661 accactggtt ggcgaggtgg acacgttgtt gaaggactgg ctggggaact tgaacaactt
     3721 cgtgcacgac tggagcatca cccacaaggt caacgtgaac catagggcct cgagggccat
     3781 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     3841 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     3901 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3961 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
     4021 gcgtcgagtg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4081 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4141 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4201 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4261 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4321 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4381 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4441 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4501 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4561 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4621 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4681 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     4741 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4801 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4861 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4921 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     4981 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5041 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5101 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5161 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5221 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5281 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5341 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5401 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5461 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5521 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5581 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5641 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5701 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5761 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5821 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5881 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5941 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6001 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6061 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6121 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6181 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6241 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
