LOCUS       sequence-402543-        7453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RUSH cargo: Str-KDEL_IRES_SBP-mCherry-VSVG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7453)
  AUTHORS   Chen Y, Gershlick DC, Park SY, Bonifacino JS
  TITLE     Segregation in the Golgi complex precedes export of endolysosomal 
            proteins in distinct transport carriers.
  JOURNAL   J Cell Biol. 2017 Dec 4;216(12):4141-4151. doi: 
            10.1083/jcb.201707172. Epub 2017 Oct 4.
  PUBMED    28978644
REFERENCE   2  (bases 1 to 7453)
  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"; doi: 
            "10.1083/jcb.201707172"; journalName: "J Cell Biol"; date: 
            "2017-12-4- 4"; volume: "216"; issue: "12"; pages: "4141-4151"
COMMENT     Sequence Label: VSVG-mCherry-RUSH
FEATURES             Location/Qualifiers
     source          1..7453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(208..999,1000..1068))
                     /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     831..850
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1069..1173)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1248..1267)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1439..1818
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1819..2022
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1973..1993
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2067..2086
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2067..2085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          2850..3079
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3062..3081
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(3151..3725,3726..3728,3729..3737)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3318..3335)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3545..3564
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     sig_peptide     3751..3810
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             3811..3921
                     /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="DEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     CDS             3931..4635
                     /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(4073..4091)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4341..4360
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             6100..6132
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     polyA_signal    6277..6398
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6314..6333)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6500..6516)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6500..6516)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6513..6535)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6524..6540
                     /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(join(6548..6554,6555..6572,6573..6578))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6593..6614
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6731..6748)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6902..7453,1..37))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6982..7001)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
       61 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
      121 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
      181 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
      241 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
      301 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
      361 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
      421 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
      481 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
      541 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
      601 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
      661 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
      721 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
      781 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
      841 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
      901 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
      961 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     1021 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     1081 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     1141 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     1201 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     1261 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     1321 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     1381 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
     1441 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     1501 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     1561 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     1621 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     1681 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     1741 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     1801 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     1861 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     1921 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     1981 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa
     2041 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagcttgg
     2101 taccgagctc ggatcgatat ctgcggccta gctagcgctt aaggcctgtt aaccggtatg
     2161 gatgtatgcg tccgtcttgc cctgtggctc ctctggggac tcctcctgca ccagggccag
     2221 agcctcagcc atagtcacag tgagaaggcg acaggaacca gctcgggcgc cggtttaaac
     2281 gaccctagca aagactcaaa agctcaggtg tccgctgccg aggctggcat tactggaaca
     2341 tggtacaatc agctcgggag cacctttatt gtgactgctg gagccgatgg agccctcacc
     2401 ggaacatacg aatctgctgt gggaaacgcc gaatcacggt acgtcctcac tggccgatac
     2461 gatagtgccc ctgccaccga cggatctggg actgccctgg gatggactgt cgcttggaaa
     2521 aacaactacc ggaatgctca ttctgccaca acatggagtg gacagtacgt gggaggcgct
     2581 gaggctagaa tcaatacaca gtggctgctc acatctggca caaccgaggc aaatgcttgg
     2641 aaatccaccc tggtgggaca tgacacattc accaaagtga aaccctccgc cgcttcaatc
     2701 gatgccgcca aaaaagccgg agtcaacaac ggcaatcctc tggatgccgt ccagcagctc
     2761 ggaccgaaag atgaactgtg agcggccgca tagataactg atccagtgtg ctggaattaa
     2821 ttcgctgtct gcgagggcca gctgttgggg tgagtactcc ctctcaaaag cgggcatgac
     2881 ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca cctggcccgc
     2941 ggtgatgcct ttgagggtgg ccgcgtccat ctggtcagaa aagacaatct ttttgttgtc
     3001 aagcttgagg tgtggcaggc ttgagatctg gccatacact tgagtgacaa tgacatccac
     3061 tttgcctttc tctccacagg tgtccactcc caggtccaac tgcaggtcga gcatgcatct
     3121 agggcggcca attccgcccc tcyccccccc cccctctccc tccccccccc ctaacgttac
     3181 tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat
     3241 attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat
     3301 tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga
     3361 agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca
     3421 gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac
     3481 acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt
     3541 caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca
     3601 ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt
     3661 aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg
     3721 ataagcttgc cacaacccgg gaggcgcgcc atgtacagga tgcaactcct gtcttgcatt
     3781 gcactaagtc ttgcacttgt cacgaattcc gacgagaaga ccactggttg gcgaggtgga
     3841 cacgttgttg aaggactggc tggggaactt gaacaacttc gtgcacgact ggagcatcac
     3901 ccacaaggtc aacgtgaacc acctgcaggt gtgagcaagg gcgaggagga taacatggcc
     3961 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     4021 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     4081 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     4141 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     4201 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     4261 gtgacccagg actcctccct acaggacggc gagttcatct acaaggtgaa gctgcgcggc
     4321 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     4381 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     4441 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     4501 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     4561 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     4621 gacgagctgt acaagggccg gccagacaag ttcaccatag tttttccaca caaccaaaaa
     4681 ggaaactgga aaaatgttcc ttctaattac cattattgcc cgtcaagctc agatttaaat
     4741 tggcataatg acttaatagg cacagcctta caagtcaaaa tgcccaagag tcacaaggct
     4801 attcaagcag acggttggat gtgtcatgct tccaaatggg tcactacttg tgatttccgc
     4861 tggtatggac cgaagtatat aacacattcc atccgatcct tcactccatc tgtagaacaa
     4921 tgcaaggaaa gcattgaaca aacgaaacaa ggaacttggc tgaatccagg cttccctcct
     4981 caaagttgtg gatatgcaac tgtgacggat gccgaagcag tgattgtcca ggtgactcct
     5041 caccatgtgc tggttgatga atacacagga gaatgggttg attcacagtt catcaacgga
     5101 aaatgcagca attacatatg ccccactgtc cataactcta caacctggca ttctgactat
     5161 aaggtcaaag ggctatgtga ttctaacctc atttccatgg acatcacctt cttctcagag
     5221 gacggagagc tatcatccct gggaaaggag ggcacagggt tcagaagtaa ctactttgct
     5281 tatgaaactg gaggcaaggc ctgcaaaatg caatactgca agcattgggg agtcagactc
     5341 ccatcaggtg tctggttcga gatggctgat aaggatctct ttgctgcagc cagattccct
     5401 gaatgcccag aagggtcaag tatctctgct ccatctcaga cctcagtgga tgtaagtcta
     5461 attcaggacg ttgagaggat cttggattat tccctctgcc aagaaacctg gagcaaaatc
     5521 agagcgggtc ttccaatctc tccagtggat ctcagctatc ttgctcctaa aaacccagga
     5581 accggtcctg ctttcaccat aatcaatggt accctaaaat actttgagac cagatacatc
     5641 agagtcgata ttgctgctcc aatcctctca agaatggtcg gaatgatcag tggaactacc
     5701 acagaaaggg aactgtggga tgactgggca ccatatgaag acgtggaaat tggacccaat
     5761 ggagttctga ggaccagttc aggatataag tttcctttat acatgattgg acatggtatg
     5821 ttggactccg atcttcatct tagctcaaag gctcaggtgt tcgaacatcc tcacattcaa
     5881 gacgctgctt cgcaacttcc tgatgatgag agtttatttt ttggtgatac tgggctatcc
     5941 aaaaatccaa tcgagcttgt agaaggttgg ttcagtagtt ggaaaagctc tattgcctct
     6001 tttttcttta tcatagggtt aatcattgga ctattcttgg ttctccgagt tggtatccat
     6061 ctttgcatta aattaaagca caccaagaaa agacagattt atacagacat agagatgaac
     6121 cgacttggaa agtagggcct cgagggccaa ttaattaact ctagataact gatcataatc
     6181 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     6241 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     6301 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6361 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtcgagtg cattctagtt
     6421 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     6481 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6541 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6601 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6661 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6721 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6781 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6841 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6901 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6961 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7021 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7081 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7141 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7201 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7261 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7321 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     7381 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7441 gtttttttgt ttg
//
