LOCUS       sequence-328226-        8204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RUSH cargo: Str-Ii_IRES_VSVG-SBP-Halo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8204)
  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 8204)
  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: Halo-VSVG-RUSH
FEATURES             Location/Qualifiers
     source          1..8204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..32)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        204..583
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        584..787
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     738..758
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     832..851
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        832..850
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1424..1450
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          2185..2414
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2397..2416
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2487..3060
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2654..2671)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2881..2900
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3087..3134,3135..4619)
                     /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             4629..4742
                     /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             4758..5642
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     polyA_signal    5793..5914
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5830..5849)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6016..6032)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6016..6032)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6029..6051)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6040..6056
                     /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(6064..6070,6071..6088,6089..6094))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6109..6130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6247..6264)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6418..7006)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6498..6517)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7177..7968,7969..8037))
                     /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     7800..7819
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8038..8142)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
       61 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
      121 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
      181 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
      241 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      301 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      361 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      421 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      481 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      541 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      601 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      661 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      721 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      781 cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
      841 tcactatagg gagacccaag cttggtaccg agctcggatc gatatcatgc accggaggag
      901 atcacgctct tgtagggagg accagaaacc tgtcaccggt gaccctagca aagactcaaa
      961 agctcaggtg tccgctgccg aggctggcat tactggaaca tggtacaatc agctcgggag
     1021 cacctttatt gtgactgctg gagccgatgg agccctcacc ggaacatacg aatctgctgt
     1081 gggaaacgcc gaatcacggt acgtcctcac tggccgatac gatagtgccc ctgccaccga
     1141 cggatctggg actgccctgg gatggactgt cgcttggaaa aacaactacc ggaatgctca
     1201 ttctgccaca acatggagtg gacagtacgt gggaggcgct gaggctagaa tcaatacaca
     1261 gtggctgctc acatctggca caaccgaggc aaatgcttgg aaatccaccc tggtgggaca
     1321 tgacacattc accaaagtga aaccctccgc cgcttcaatc gatgccgcca aaaaagccgg
     1381 agtcaacaac ggcaatcctc tggatgccgt ccagcaggtc gactatccgt acgacgtacc
     1441 agactacgca gtcggaccga tggacgatca gagggacctc attagcaaca acgaacagct
     1501 gcctatgctg ggacggcgac ctggagcccc tgaatccaaa tgctctaggg gagcactgta
     1561 cactggcttc tccattctcg tgacactgct gctggccggg caggctacta ctgcttactt
     1621 cctgtaccag cagcaggggc ggctggacaa actcactgtg acatctcaga acctccagct
     1681 ggaaaatctg aggatgaaac tgcccaaacc ccctaaaccc gtgtccaaaa tgaggatggc
     1741 cacacctctg ctcatgcagg cactgccaat gggagccctg ccccaggggc ccatgcagaa
     1801 tgccaccaag tatggcaaca tgacagagga ccatgtgatg cacctgctcc agaatgctga
     1861 ccccctgaag gtgtacccgc cactgaaggg gagcttcccg gagaacctga gacaccttaa
     1921 gaacaccatg gagaccatag actggaaggt ctttgagagc tggatgcacc attggctcct
     1981 gtttgaaatg agcaggcact ccttggagca aaagcccact gacgctccac cgaaagagtc
     2041 actggaactg gaggacccgt cttctgggct gggtgtgacc aagcaggatc tgggcccagt
     2101 ccccatgtga ggatccgcgg ccgcatagat aactgatcca gtgtgctgga attaattcgc
     2161 tgtctgcgag ggccagctgt tggggtgagt actccctctc aaaagcgggc atgacttctg
     2221 cgctaagatt gtcagtttcc aaaaacgagg aggatttgat attcacctgg cccgcggtga
     2281 tgcctttgag ggtggccgcg tccatctggt cagaaaagac aatctttttg ttgtcaagct
     2341 tgaggtgtgg caggcttgag atctggccat acacttgagt gacaatgaca tccactttgc
     2401 ctttctctcc acaggtgtcc actcccaggt ccaactgcag gtcgagcatg catctagggc
     2461 ggccaattcc gcccctctcc cccccccccn tctccctccc ccccccctaa cgttactggc
     2521 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     2581 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     2641 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     2701 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     2761 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     2821 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     2881 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     2941 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     3001 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     3061 gcttgccaca acccgggagg cgcgccatga agtgcctttt gtacttagcc tttttattca
     3121 ttggggtgaa ttgcaagttc accatagttt ttccacacaa ccaaaaagga aactggaaaa
     3181 atgttccttc taattaccat tattgcccgt caagctcaga tttaaattgg cataatgact
     3241 taataggcac agccttacaa gtcaaaatgc ccaagagtca caaggctatt caagcagacg
     3301 gttggatgtg tcatgcttcc aaatgggtca ctacttgtga tttccgctgg tatggaccga
     3361 agtatataac acattccatc cgatccttca ctccatctgt agaacaatgc aaggaaagca
     3421 ttgaacaaac gaaacaagga acttggctga atccaggctt ccctcctcaa agttgtggat
     3481 atgcaactgt gacggatgcc gaagcagtga ttgtccaggt gactcctcac catgtgctgg
     3541 ttgatgaata cacaggagaa tgggttgatt cacagttcat caacggaaaa tgcagcaatt
     3601 acatatgccc cactgtccat aactctacaa cctggcattc tgactataag gtcaaagggc
     3661 tatgtgattc taacctcatt tccatggaca tcaccttctt ctcagaggac ggagagctat
     3721 catccctggg aaaggagggc acagggttca gaagtaacta ctttgcttat gaaactggag
     3781 gcaaggcctg caaaatgcaa tactgcaagc attggggagt cagactccca tcaggtgtct
     3841 ggttcgagat ggctgataag gatctctttg ctgcagccag attccctgaa tgcccagaag
     3901 ggtcaagtat ctctgctcca tctcagacct cagtggatgt aagtctaatt caggacgttg
     3961 agaggatctt ggattattcc ctctgccaag aaacctggag caaaatcaga gcgggtcttc
     4021 caatctctcc agtggatctc agctatcttg ctcctaaaaa cccaggaacc ggtcctgctt
     4081 tcaccataat caatggtacc ctaaaatact ttgagaccag atacatcaga gtcgatattg
     4141 ctgctccaat cctctcaaga atggtcggaa tgatcagtgg aactaccaca gaaagggaac
     4201 tgtgggatga ctgggcacca tatgaagacg tggaaattgg acccaatgga gttctgagga
     4261 ccagttcagg atataagttt cctttataca tgattggaca tggtatgttg gactccgatc
     4321 ttcatcttag ctcaaaggct caggtgttcg aacatcctca cattcaagac gctgcttcgc
     4381 aacttcctga tgatgagagt ttattttttg gtgatactgg gctatccaaa aatccaatcg
     4441 agcttgtaga aggttggttc agtagttgga aaagctctat tgcctctttt ttctttatca
     4501 tagggttaat cattggacta ttcttggttc tccgagttgg tatccatctt tgcattaaat
     4561 taaagcacac caagaaaaga cagatttata cagacataga gatgaaccga cttggaaagc
     4621 tgaattccat ggacgagaag accactggtt ggcgaggtgg acacgttgtt gaaggactgg
     4681 ctggggaact tgaacaactt cgtgcacgac tggagcatca cccacaaggt caacgtgaac
     4741 cacctgcagg tggatccgaa atcggtactg gctttccatt cgacccccat tatgtggaag
     4801 tcctgggcga gcgcatgcac tacgtcgatg ttggtccgcg cgatggcacc cctgtgctgt
     4861 tcctgcacgg taacccgacc tcctcctacg tgtggcgcaa catcatcccg catgttgcac
     4921 cgacccatcg ctgcattgct ccagacctga tcggtatggg caaatccgac aaaccagacc
     4981 tgggttattt cttcgacgac cacgtccgct tcatggatgc cttcatcgaa gccctgggtc
     5041 tggaagaggt cgtcctggtc attcacgact ggggctccgc tctgggtttc cactgggcca
     5101 agcgcaatcc agagcgcgtc aaaggtattg catttatgga gttcatccgc cctatcccga
     5161 cctgggacga atggccagaa tttgcccgcg agaccttcca ggccttccgc accaccgacg
     5221 tcggccgcaa gctgatcatc gatcagaacg tttttatcga gggtacgctg ccgatgggtg
     5281 tcgtccgccc gctgactgaa gtcgagatgg accattaccg cgagccgttc ctgaatcctg
     5341 ttgaccgcga gccactgtgg cgcttcccaa acgagctgcc aatcgccggt gagccagcga
     5401 acatcgtcgc gctggtcgaa gaatacatgg actggctgca ccagtcccct gtcccgaagc
     5461 tgctgttctg gggcacccca ggcgttctga tcccaccggc cgaagccgct cgcctggcca
     5521 aaagcctgcc taactgcaag gctgtggaca tcggcccggg tctgaatctg ctgcaagaag
     5581 acaacccgga cctgatcggc agcgagatcg cgcgctggct gtctactctg gagatttccg
     5641 gccggcctta aggcctcgag ggccaattaa ttaactctag ataactgatc ataatcagcc
     5701 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     5761 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     5821 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5881 gttgtggttt gtccaaactc atcaatgtat cttaacgcgt cgagtgcatt ctagttgtgg
     5941 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     6001 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6061 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6121 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6181 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6241 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6301 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6361 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6421 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6481 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6541 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6601 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6661 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6721 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6781 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6841 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6901 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6961 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7021 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7081 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7141 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7201 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7261 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7321 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7381 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7441 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7501 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7561 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7621 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7681 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7741 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7801 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7861 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7921 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7981 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8041 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8101 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8161 gccacctgac gtcgacggat cgggagatct cccgatcccc tatg
//
