LOCUS       sequence-328225-        8203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RUSH cargo: Str-KDEL_IRES_Transferrin Receptor (TfR)-SBP-mApple.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8203)
  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 8203)
  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-TfR-RUSH
FEATURES             Location/Qualifiers
     source          1..8203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     225..244
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             complement(1370..1381)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2720..2833
                     /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             2843..3550
                     /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     3256..3275
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3707..3828
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3744..3763)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3930..3946)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3930..3946)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3943..3965)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3954..3970
                     /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(3978..3984,3985..4002,4003..4008))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4023..4044
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4161..4178)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4332..4920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4412..4431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5091..5882,5883..5951))
                     /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     5714..5733
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5952..6056)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6131..6150)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6322..6701
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6702..6905
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6856..6876
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6950..6969
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6950..6968
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          7733..7962
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     7945..7964
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(8034..8203,1..405,406..408,409..417)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(join(8201..8203,1..15))
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
ORIGIN
        1 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
       61 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
      121 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
      181 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
      241 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
      301 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
      361 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataagcttgc cacaacccgg
      421 gaggcgcgcc atgatggatc aagctagatc agcattctct aacttgtttg gtggagaacc
      481 attgtcatat acccggttca gcctggctcg gcaagtagat ggcgataaca gtcatgtgga
      541 gatgaaactt gctgtagatg aagaagaaaa tgctgacaat aacacaaagg ccaatgtcac
      601 aaaaccaaaa aggtgtagtg gaagtatctg ctatgggact attgctgtga tcgtcttttt
      661 cttgattgga tttatgattg gctacttggg ctattgtaaa ggggtagaac caaaaactga
      721 gtgtgagaga ctggcaggaa ccgagtctcc agtgagggag gagccaggag aggacttccc
      781 tgcagcacgt cgcttatatt gggatgacct gaagagaaag ttgtcggaga aactggacag
      841 cacagacttc accagcacca tcaagctgct gaatgaaaat tcatatgtcc ctcgtgaggc
      901 tggatctcaa aaagatgaaa atcttgcgtt gtatgttgaa aatcaatttc gtgaatttaa
      961 actcagcaaa gtctggcgtg atcaacattt tgttaagatt caggtcaaag acagcgctca
     1021 aaactcggtg atcatagttg ataagaacgg tagacttgtt tacctggtgg agaatcctgg
     1081 gggttatgtg gcgtatagta aggctgcaac agttactggt aaactggtcc atgctaattt
     1141 tggtactaaa aaagattttg aggatttata cactcctgtg aatggatcta tagtgattgt
     1201 cagagcaggg aaaatcacct ttgcagaaaa ggttgcaaat gctgaaagct taaatgcaat
     1261 tggtgtgttg atatacatgg accagactaa atttcccatt gttaacgcag aactttcatt
     1321 ctttggacat gctcatctgg ggacaggtga cccttacaca cctggattcc cttccttcaa
     1381 tcacactcag tttccaccat ctcggtcatc aggattgcct aatatacctg tccagacaat
     1441 ctccagagct gctgcagaaa agctgtttgg gaatatggaa ggagactgtc cctctgactg
     1501 gaaaacagac tctacatgta ggatggtaac ctcagaaagc aagaatgtga agctcactgt
     1561 gagcaatgtg ctgaaagaga taaaaattct taacatcttt ggagttatta aaggctttgt
     1621 agaaccagat cactatgttg tagttggggc ccagagagat gcatggggcc ctggagctgc
     1681 aaaatccggt gtaggcacag ctctcctatt gaaacttgcc cagatgttct cagatatggt
     1741 cttaaaagat gggtttcagc ccagcagaag cattatcttt gccagttgga gtgctggaga
     1801 ctttggatcg gttggtgcca ctgaatggct agagggatac ctttcgtccc tgcatttaaa
     1861 ggctttcact tatattaatc tggataaagc ggttcttggt accagcaact tcaaggtttc
     1921 tgccagccca ctgttgtata cgcttattga gaaaacaatg caaaatgtga agcatccggt
     1981 tactgggcaa tttctatatc aggacagcaa ctgggccagc aaagttgaga aactcacttt
     2041 agacaatgct gctttccctt tccttgcata ttctggaatc ccagcagttt ctttctgttt
     2101 ttgcgaggac acagattatc cttatttggg taccaccatg gacacctata aggaactgat
     2161 tgagaggatt cctgagttga acaaagtggc acgagcagct gcagaggtcg ctggtcagtt
     2221 cgtgattaaa ctaacccatg atgttgaatt gaacctggac tatgagaggt acaacagcca
     2281 actgctttca tttgtgaggg atctgaacca atacagagca gacataaagg aaatgggcct
     2341 gagtttacag tggctgtatt ctgctcgtgg agacttcttc cgtgctactt ccagactaac
     2401 aacagatttc gggaatgctg agaaaacaga cagatttgtc atgaagaaac tcaatgatcg
     2461 tgtcatgaga gtggagtatc acttcctctc tccctacgta tctccaaaag agtctccttt
     2521 ccgacatgtc ttctggggct ccggctctca cacgctgcca gctttactgg agaacttgaa
     2581 actgcgtaaa caaaataacg gtgcttttaa tgaaacgctg ttcagaaacc agttggctct
     2641 agctacttgg actattcagg gagctgcaaa tgccctctct ggtgacgttt gggacattga
     2701 caatgagttt ctgaattcca tggacgagaa gaccactggt tggcgaggtg gacacgttgt
     2761 tgaaggactg gctggggaac ttgaacaact tcgtgcacga ctggagcatc acccacaagg
     2821 tcaacgtgaa ccacctgcag gtatggtgag caagggcgag gagaataaca tggccatcat
     2881 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
     2941 gatcgagggc gagggcgagg gccgccccta cgaggccttt cagaccgcta agctgaaggt
     3001 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     3061 ctccaaggtc tacattaagc acccagccga catccccgac tacttcaagc tgtccttccc
     3121 cgagggcttc aggtgggagc gcgtgatgaa cttcgaggac ggcggcatta ttcacgttaa
     3181 ccaggactcc tccctgcaag acggcgtgtt catctacaag gtgaagctgc gcggcaccaa
     3241 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccgagga
     3301 gcggatgtac cccgaggacg gcgccctgaa gagcgagatc aagaagaggc tgaagctgaa
     3361 ggacggcggc cactacgccg ccgaggtcaa gaccacctac aaggccaaga agcccgtgca
     3421 gctgcccggc gcctacatcg tcgacatcaa gttggacatc gtgtcccaca acgaggacta
     3481 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     3541 gctgtacaag tccggccggc cttaaggcct cgagggccaa ttaattaact ctagataact
     3601 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     3661 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3721 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3781 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtcgagtg
     3841 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     3901 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     3961 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4021 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4081 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4141 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4201 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4261 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4321 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4381 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4441 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4501 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4561 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4621 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4681 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4741 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4801 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4861 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4921 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4981 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5041 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5101 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5161 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5221 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5281 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5341 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5401 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5461 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5521 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5581 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5641 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5701 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5761 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5821 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5881 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5941 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6001 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6061 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     6121 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     6181 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     6241 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     6301 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
     6361 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     6421 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     6481 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     6541 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     6601 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     6661 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     6721 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     6781 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     6841 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     6901 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
     6961 actataggga gacccaagct tggtaccgag ctcggatcga tatctgcggc ctagctagcg
     7021 cttaaggcct gttaaccggt atggatgtat gcgtccgtct tgccctgtgg ctcctctggg
     7081 gactcctcct gcaccagggc cagagcctca gccatagtca cagtgagaag gcgacaggaa
     7141 ccagctcggg cgccggttta aacgacccta gcaaagactc aaaagctcag gtgtccgctg
     7201 ccgaggctgg cattactgga acatggtaca atcagctcgg gagcaccttt attgtgactg
     7261 ctggagccga tggagccctc accggaacat acgaatctgc tgtgggaaac gccgaatcac
     7321 ggtacgtcct cactggccga tacgatagtg cccctgccac cgacggatct gggactgccc
     7381 tgggatggac tgtcgcttgg aaaaacaact accggaatgc tcattctgcc acaacatgga
     7441 gtggacagta cgtgggaggc gctgaggcta gaatcaatac acagtggctg ctcacatctg
     7501 gcacaaccga ggcaaatgct tggaaatcca ccctggtggg acatgacaca ttcaccaaag
     7561 tgaaaccctc cgccgcttca atcgatgccg ccaaaaaagc cggagtcaac aacggcaatc
     7621 ctctggatgc cgtccagcag ctcggaccga aagatgaact gtgagcggcc gcatagataa
     7681 ctgatccagt gtgctggaat taattcgctg tctgcgaggg ccagctgttg gggtgagtac
     7741 tccctctcaa aagcgggcat gacttctgcg ctaagattgt cagtttccaa aaacgaggag
     7801 gatttgatat tcacctggcc cgcggtgatg cctttgaggg tggccgcgtc catctggtca
     7861 gaaaagacaa tctttttgtt gtcaagcttg aggtgtggca ggcttgagat ctggccatac
     7921 acttgagtga caatgacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc
     7981 aactgcaggt cgagcatgca tctagggcgg ccaattccgc ccctctcccc ccccccctct
     8041 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     8101 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     8161 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctc
//
