LOCUS       sequence-402548-        6721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RUSH cargo: Str-KDEL_IRES_SBP-mCherry-CD-MPR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6721)
  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 6721)
  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: CD-MPR-mCherry-RUSH
FEATURES             Location/Qualifiers
     source          1..6721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(125..144)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          127..356
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     complement(1120..1139)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1121..1139)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1184..1387)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1213..1233)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1388..1767
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1939..1958
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2033..2137
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2138..2206,2207..2998)
                     /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     complement(2356..2375)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3169..3757
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3658..3677
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3911..3928
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4045..4066
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4081..4086,4087..4104,4105..4111)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4119..4135
                     /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)."
     primer_bind     4124..4146
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4143..4159
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4143..4159
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4261..4382
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4326..4345
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5292..5996)
                     /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(5567..5586)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5836..5854
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(6006..6116)
                     /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"
     sig_peptide     complement(6117..6176)
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     misc_feature    join(6190..6198,6199..6201,6202..6721,1..55)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6363..6382)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     6592..6609
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
ORIGIN
        1 gccttattcc aagcggcttc ggccagtaac gttagggggg ggggagggag aggggggggg
       61 ggagaggggc ggaattggcc gccctagatg catgctcgac ctgcagttgg acctgggagt
      121 ggacacctgt ggagagaaag gcaaagtgga tgtcattgtc actcaagtgt atggccagat
      181 ctcaagcctg ccacacctca agcttgacaa caaaaagatt gtcttttctg accagatgga
      241 cgcggccacc ctcaaaggca tcaccgcggg ccaggtgaat atcaaatcct cctcgttttt
      301 ggaaactgac aatcttagcg cagaagtcat gcccgctttt gagagggagt actcacccca
      361 acagctggcc ctcgcagaca gcgaattaat tccagcacac tggatcagtt atctatgcgg
      421 ccgctcacag ttcatctttc ggtccgagct gctggacggc atccagagga ttgccgttgt
      481 tgactccggc ttttttggcg gcatcgattg aagcggcgga gggtttcact ttggtgaatg
      541 tgtcatgtcc caccagggtg gatttccaag catttgcctc ggttgtgcca gatgtgagca
      601 gccactgtgt attgattcta gcctcagcgc ctcccacgta ctgtccactc catgttgtgg
      661 cagaatgagc attccggtag ttgtttttcc aagcgacagt ccatcccagg gcagtcccag
      721 atccgtcggt ggcaggggca ctatcgtatc ggccagtgag gacgtaccgt gattcggcgt
      781 ttcccacagc agattcgtat gttccggtga gggctccatc ggctccagca gtcacaataa
      841 aggtgctccc gagctgattg taccatgttc cagtaatgcc agcctcggca gcggacacct
      901 gagcttttga gtctttgcta gggtcgttta aaccggcgcc cgagctggtt cctgtcgcct
      961 tctcactgtg actatggctg aggctctggc cctggtgcag gaggagtccc cagaggagcc
     1021 acagggcaag acggacgcat acatccatac cggttaacag gccttaagcg ctagctaggc
     1081 cgcagatatc gatccgagct cggtaccaag cttgggtctc cctatagtga gtcgtattaa
     1141 tttcgataag ccagtaagca gtgggttctc tagttagcca gagagctctg cttatataga
     1201 cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt
     1261 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga
     1321 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
     1381 gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     1441 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     1501 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     1561 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     1621 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     1681 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
     1741 attaataact agtcaataat caatgtcaac gcgtatatct ggcccgtaca tcgcgaagca
     1801 gcgcaaaacg cctaacccta agcagattct tcatgcaatt gtcggtcaag ccttgccttg
     1861 ttgtagctta aattttgctc gcgcactact cagcgacctc caacacacaa gcagggagca
     1921 gatactggct taactatgcg gcatcagagc agattgtact gagagtgcac cataggggat
     1981 cgggagatct cccgatccgt cgacgtcagg tggcactttt cggggaaatg tgcgcggaac
     2041 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2101 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     2161 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     2221 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     2281 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     2341 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     2401 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     2461 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     2521 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     2581 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     2641 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     2701 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     2761 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     2821 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     2881 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     2941 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     3001 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     3061 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     3121 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3181 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3241 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3301 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3361 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3421 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3481 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3541 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3601 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3661 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3721 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3781 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3841 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3901 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     3961 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     4021 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     4081 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     4141 cacaggaaac agctatgacc atgattacgc caagctctag ctagaggtcg acggtataca
     4201 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca ctcgacgcgt
     4261 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     4321 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     4381 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     4441 tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcag ttatctagag
     4501 ttaattaatt ggccctcgag gccctacatt ggtaataaat ggtcatccct ttcttctgac
     4561 tcctccccca gctggtcatc ccccacacca cgatatgctg caggcacatt tcgaggttta
     4621 gaacggcaga caaagtcaca gccatctgct accaggttgc caagatcctg ccagaaggct
     4681 aagtggggaa actgctccat tcctttggct cccactacca gtcgctggta taggaacccc
     4741 ccaacaacat aaacagcaac cagtgatgca aacgtgacaa gtaagatgga acccacactg
     4801 aggtgggaga tctctggtga acaggccagg ctgctatcca tctcaaagag gtagaaacaa
     4861 tcttggactt tgccacgctc ctcagacaca gggttaaaat tgtccgctag ggtgtgtcga
     4921 ttgcaggaga tcatcaccac tgcacgacgc tgctccttgc cacagtggtt gtcatattca
     4981 tcaccccctt tatagatcag catgatccaa ttacttccgt tgaagatgtg agtctcgttg
     5041 agtctcccta ccactgtctc cttcccatta cttttgttga tttgcaccag gcctgcccca
     5101 gaagtgtggt tgccagcttc ccggcacacc ctgaagatgt agatgtatgt gtctgaaccc
     5161 tggcccacag tgctctcaaa gcttttatta aacagtggtt tcagcctctt cactagagcc
     5221 aactctttct ctgactctct acccttttct cctaccaagt cgcaagtttt ttcttctgtg
     5281 tctggccggc ccttgtacag ctcgtccatg ccgccggtgg agtggcggcc ctcggcgcgt
     5341 tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg tgatgtccaa cttgatgttg
     5401 acgttgtagg cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt cttgacctca
     5461 gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct tgatctcgcc cttcagggcg
     5521 ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc ccatggtctt cttctgcatt
     5581 acggggccgt cggaggggaa gttggtgccg cgcagcttca ccttgtagat gaactcgccg
     5641 tcctgtaggg aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg
     5701 cgctcccact tgaagccctc ggggaaggac agcttcaagt agtcggggat gtcggcgggg
     5761 tgcttcacgt aggccttgga gccgtacatg aactgagggg acaggatgtc ccaggcgaag
     5821 ggcagggggc cacccttggt caccttcagc ttggcggtct gggtgccctc gtaggggcgg
     5881 ccctcgccct cgccctcgat ctcgaactcg tggccgttca cggagccctc catgtgcacc
     5941 ttgaagcgca tgaactcctt gatgatggcc atgttatcct cctcgccctt gctcacacct
     6001 gcaggtggtt cacgttgacc ttgtgggtga tgctccagtc gtgcacgaag ttgttcaagt
     6061 tccccagcca gtccttcaac aacgtgtcca cctcgccaac cagtggtctt ctcgtcggaa
     6121 ttcgtgacaa gtgcaagact tagtgcaatg caagacagga gttgcatcct gtacatggcg
     6181 cgcctcccgg gttgtggcaa gcttatcatc gtgtttttca aaggaaaacc acgtccccgt
     6241 ggttcggggg gcctagacgt ttttttaacc tcgactaaac acatgtaaag catgtgcacc
     6301 gaggccccag atcagatccc atacaatggg gtaccttctg ggcatccttc agccccttgt
     6361 tgaatacgct tgaggagagc catttgactc tttccacaac tatccaactc acaacgtggc
     6421 actggggttg tgccgccttt gcaggtgtat cttatacacg tggcttttgg ccgcagaggc
     6481 acctgtcgcc aggtgggggg ttccgctgcc tgcaaagggt cgctacagac gttgtttgtc
     6541 ttcaagaagc ttccagagga actgcttcct tcacgacatt caacagacct tgcattcctt
     6601 tggcgagagg ggaaagaccc ctaggaatgc tcgtcaagaa gacagggcca ggtttccggg
     6661 ccctcacatt gccaaaagac ggcaatatgg tggaaaataa catatagaca aacgcacacc
     6721 g
//
