LOCUS       sequence-328223-        6058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RUSH cargo: Str-KDEL_IRES_SBP-mApple-GPI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6058)
  AUTHORS   Weigel AV, Chang CL, Shtengel G, Xu CS, Hoffman DP, Freeman M, Iyer 
            N, Aaron J, Khuon S, Bogovic J, Qiu W, Hess HF, Lippincott-Schwartz 
            J
  TITLE     ER-to-Golgi protein delivery through an interwoven, tubular network 
            extending from ER.
  JOURNAL   Cell. 2021 Apr 29;184(9):2412-2429.e16. doi: 
            10.1016/j.cell.2021.03.035. Epub 2021 Apr 13.
  PUBMED    33852913
REFERENCE   2  (bases 1 to 6058)
  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: 
            "10.1016/j.cell.2021.03.035"; journalName: "Cell"; date: 
            "2021-04-29- 29"; volume: "184"; issue: "9"; pages: "2412-2429.e16"
COMMENT     Sequence Label: mApple-GPI-RUSH
FEATURES             Location/Qualifiers
     source          1..6058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        157..536
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        537..740
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     691..711
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     785..804
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        785..803
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          1568..1797
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1780..1799
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(1869..2443,2444..2446,2447..2455)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2036..2053)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2263..2282
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     sig_peptide     2469..2528
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             2529..2639
                     /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             2649..3356
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK
                     SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     3062..3081
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3600..3721
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3637..3656)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3823..3839)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3823..3839)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3836..3858)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3847..3863
                     /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(3871..3877,3878..3895,3896..3901))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3916..3937
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4054..4071)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4225..4813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4305..4324)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4984..5775,5776..5844))
                     /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     5607..5626
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5845..5949)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6024..6043)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
       61 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
      121 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt
      181 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      241 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      301 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      361 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      421 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      481 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      541 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      601 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      661 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      721 aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg
      781 aaattaatac gactcactat agggagaccc aagcttggta ccgagctcgg atcgatatct
      841 gcggcctagc tagcgcttaa ggcctgttaa ccggtatgga tgtatgcgtc cgtcttgccc
      901 tgtggctcct ctggggactc ctcctgcacc agggccagag cctcagccat agtcacagtg
      961 agaaggcgac aggaaccagc tcgggcgccg gtttaaacga ccctagcaaa gactcaaaag
     1021 ctcaggtgtc cgctgccgag gctggcatta ctggaacatg gtacaatcag ctcgggagca
     1081 cctttattgt gactgctgga gccgatggag ccctcaccgg aacatacgaa tctgctgtgg
     1141 gaaacgccga atcacggtac gtcctcactg gccgatacga tagtgcccct gccaccgacg
     1201 gatctgggac tgccctggga tggactgtcg cttggaaaaa caactaccgg aatgctcatt
     1261 ctgccacaac atggagtgga cagtacgtgg gaggcgctga ggctagaatc aatacacagt
     1321 ggctgctcac atctggcaca accgaggcaa atgcttggaa atccaccctg gtgggacatg
     1381 acacattcac caaagtgaaa ccctccgccg cttcaatcga tgccgccaaa aaagccggag
     1441 tcaacaacgg caatcctctg gatgccgtcc agcagctcgg accgaaagat gaactgtgag
     1501 cggccgcata gataactgat ccagtgtgct ggaattaatt cgctgtctgc gagggccagc
     1561 tgttggggtg agtactccct ctcaaaagcg ggcatgactt ctgcgctaag attgtcagtt
     1621 tccaaaaacg aggaggattt gatattcacc tggcccgcgg tgatgccttt gagggtggcc
     1681 gcgtccatct ggtcagaaaa gacaatcttt ttgttgtcaa gcttgaggtg tggcaggctt
     1741 gagatctggc catacacttg agtgacaatg acatccactt tgcctttctc tccacaggtg
     1801 tccactccca ggtccaactg caggtcgagc atgcatctag ggcggccaat tccgcccctc
     1861 tccccccccc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
     1921 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     1981 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     2041 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     2101 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     2161 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     2221 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     2281 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     2341 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     2401 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aagcttgcca caacccggga
     2461 ggcgcgccat gtacaggatg caactcctgt cttgcattgc actaagtctt gcacttgtca
     2521 cgaattccga cgagaagacc actggttggc gaggtggaca cgttgttgaa ggactggctg
     2581 gggaacttga acaacttcgt gcacgactgg agcatcaccc acaaggtcaa cgtgaaccac
     2641 ctgcaggtat ggtgagcaag ggcgaggaga ataacatggc catcatcaag gagttcatgc
     2701 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     2761 gcgagggccg cccctacgag gcctttcaga ccgctaagct gaaggtgacc aagggtggcc
     2821 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggtctaca
     2881 ttaagcaccc agccgacatc cccgactact tcaagctgtc cttccccgag ggcttcaggt
     2941 gggagcgcgt gatgaacttc gaggacggcg gcattattca cgttaaccag gactcctccc
     3001 tgcaagacgg cgtgttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     3061 gccccgtaat gcagaagaag accatgggct gggaggcctc cgaggagcgg atgtaccccg
     3121 aggacggcgc cctgaagagc gagatcaaga agaggctgaa gctgaaggac ggcggccact
     3181 acgccgccga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     3241 acatcgtcga catcaagttg gacatcgtgt cccacaacga ggactacacc atcgtggaac
     3301 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtccg
     3361 gccggccact ggaaaatggc ggaacctctc tgtctgaaaa aactgtgctg ctgctggtga
     3421 ctccctttct ggccgctgct tggtccctcc acccatgagg cctcgagggc caattaatta
     3481 actctagata actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     3541 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     3601 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3661 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3721 aacgcgtcga gtgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     3781 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     3841 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3901 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3961 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4021 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4081 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4141 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4201 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4261 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4321 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4381 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4441 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4501 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4561 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4621 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4681 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4741 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4801 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4861 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4921 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4981 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5041 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5101 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5161 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5221 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5281 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5341 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5401 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5461 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5521 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5581 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5641 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5701 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5761 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5821 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5881 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5941 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     6001 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagt
//
