LOCUS       sequence-401139-        7126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RUSH cargo: Str-KDEL_IRES_SBP-mCherry-Lamp1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7126)
  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 7126)
  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: Lamp1-mCherry-RUSH
FEATURES             Location/Qualifiers
     source          1..7126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     13..30
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(267..286)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          269..498
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     complement(1262..1281)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1263..1281)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1326..1529)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1355..1375)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1530..1909
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2081..2100
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2175..2279
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2280..2348,2349..3140)
                     /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(2498..2517)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3311..3899
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3800..3819
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4053..4070
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4187..4208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4223..4228,4229..4246,4247..4253)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4261..4277
                     /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     4266..4288
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4285..4301
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4285..4301
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4403..4524
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4468..4487
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5839..6543)
                     /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(6114..6133)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6383..6401
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(6553..6663)
                     /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(6664..6723)
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     misc_feature    join(6750..7126,1..197)
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6910..6929)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
ORIGIN
        1 tcaacagacc ttgcattcct ttggcgagag gggaaagacc cctaggaatg ctcgtcaaga
       61 agacagggcc aggtttccgg gccctcacat tgccaaaaga cggcaatatg gtggaaaata
      121 acatatagac aaacgcacac cggccttatt ccaagcggct tcggccagta acgttagggg
      181 gggggnaggg agaggggggg ggggagaggg gcggaattgg ccgccctaga tgcatgctcg
      241 acctgcagtt ggacctggga gtggacacct gtggagagaa aggcaaagtg gatgtcattg
      301 tcactcaagt gtatggccag atctcaagcc tgccacacct caagcttgac aacaaaaaga
      361 ttgtcttttc tgaccagatg gacgcggcca ccctcaaagg catcaccgcg ggccaggtga
      421 atatcaaatc ctcctcgttt ttggaaactg acaatcttag cgcagaagtc atgcccgctt
      481 ttgagaggga gtactcaccc caacagctgg ccctcgcaga cagcgaatta attccagcac
      541 actggatcag ttatctatgc ggccgctcac agttcatctt tcggtccgag ctgctggacg
      601 gcatccagag gattgccgtt gttgactccg gcttttttgg cggcatcgat tgaagcggcg
      661 gagggtttca ctttggtgaa tgtgtcatgt cccaccaggg tggatttcca agcatttgcc
      721 tcggttgtgc cagatgtgag cagccactgt gtattgattc tagcctcagc gcctcccacg
      781 tactgtccac tccatgttgt ggcagaatga gcattccggt agttgttttt ccaagcgaca
      841 gtccatccca gggcagtccc agatccgtcg gtggcagggg cactatcgta tcggccagtg
      901 aggacgtacc gtgattcggc gtttcccaca gcagattcgt atgttccggt gagggctcca
      961 tcggctccag cagtcacaat aaaggtgctc ccgagctgat tgtaccatgt tccagtaatg
     1021 ccagcctcgg cagcggacac ctgagctttt gagtctttgc tagggtcgtt taaaccggcg
     1081 cccgagctgg ttcctgtcgc cttctcactg tgactatggc tgaggctctg gccctggtgc
     1141 aggaggagtc cccagaggag ccacagggca agacggacgc atacatccat accggttaac
     1201 aggccttaag cgctagctag gccgcagata tcgatccgag ctcggtacca agcttgggtc
     1261 tccctatagt gagtcgtatt aatttcgata agccagtaag cagtgggttc tctagttagc
     1321 cagagagctc tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat
     1381 ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc
     1441 ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc
     1501 ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg
     1561 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
     1621 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
     1681 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
     1741 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
     1801 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
     1861 aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtca acgcgtatat
     1921 ctggcccgta catcgcgaag cagcgcaaaa cgcctaaccc taagcagatt cttcatgcaa
     1981 ttgtcggtca agccttgcct tgttgtagct taaattttgc tcgcgcacta ctcagcgacc
     2041 tccaacacac aagcagggag cagatactgg cttaactatg cggcatcaga gcagattgta
     2101 ctgagagtgc accatagggg atcgggagat ctcccgatcc gtcgacgtca ggtggcactt
     2161 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2221 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2281 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2341 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     2401 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2461 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2521 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2581 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2641 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2701 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2761 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2821 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2881 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     2941 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3001 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3061 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3121 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3181 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3241 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3301 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3361 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3421 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3481 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3541 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3601 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3661 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3721 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3781 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3841 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3901 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3961 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     4021 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     4081 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     4141 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     4201 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     4261 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
     4321 agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac
     4381 aactagaatg cactcgacgc gttaagatac attgatgagt ttggacaaac cacaactaga
     4441 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     4501 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     4561 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
     4621 gattatgatc agttatctag agttaattaa ttggccctcg aggccctaga tggtctgata
     4681 gcccgcgtga ctcctcttcc tgccgatgag gtaggcgatg aggacgatga ggaccagccc
     4741 tgccagggcc ccgcccacag caatggggat cagcatgttg ttaccgtcct gtacacactc
     4801 ttccacagac ccaaacctgt cactttctac cctgaaagcc tggacttgca cgctgaagac
     4861 attgagggcg agcgccttgc tgacaaagat gtgctcctca ctgttgcact tgtatgagtt
     4921 gccgacactg gcctgaagag ctttcaggga atagttggag gtgctgaacg tgggctctat
     4981 ggcatcagga agagtcatgt tcaactgaac tccttgcagg aaaaacaggc tagaagtggc
     5041 attcatccca aactgcagct ccaggactct gctcttgttc cccaccttca gggtcaccaa
     5101 ctgggcaccg caagtcccac tatatttgtc acttgggttg atgttgaatg ctctggtcac
     5161 agtcgtgttg tccttcttca tgtaggtgat gttgagttgc agtgccatag aggccagcag
     5221 gcaggttcca ttgtcaccag tcacattgta cttggacaca ctggggtttg tgggcacaag
     5281 tggtggtgag gggctgggtg gcccagtagt tggggaaggt tgatcctgtg ggcagcgtgt
     5341 ctcttccttg ctgaagttgc tactcggcag gtaggcctgg atagtagcgt cccagagcac
     5401 aatggtcaca ttcttcatgt agaccctgat gtcgctgaca catcggtatg ttttgttgat
     5461 gtctgccttg atgtcagttg tggaatccac agtgtcgggc cctttggagc tggcattggg
     5521 aaagaattgt gtgtctgaca ggttatatgt gaaatacatg tgctggacac tgtaacgtgt
     5581 tgtgtttttt gtgaaggtga gtttcagtaa atatccttct ccaaaggtga ttgcgagggt
     5641 gggctcagaa gcattctttt caccacaaga gctgctattc ttcaggactt ctgcagaggc
     5701 tggcagggtc atattcgaga ccttagaaac atgtccagcc tcataggtgg tcagaaagga
     5761 ggcagagaag ctggccatta tacacgctgt gccgttgttg tctttcacct cgaacagtgc
     5821 tggggcgtct ggccggccct tgtacagctc gtccatgccg ccggtggagt ggcggccctc
     5881 ggcgcgttcg tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccaactt
     5941 gatgttgacg ttgtaggcgc cgggcagctg cacgggcttc ttggccttgt aggtggtctt
     6001 gacctcagcg tcgtagtggc cgccgtcctt cagcttcagc ctctgcttga tctcgccctt
     6061 cagggcgccg tcctcggggt acatccgctc ggaggaggcc tcccagccca tggtcttctt
     6121 ctgcattacg gggccgtcgg aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa
     6181 ctcgccgtcc tgtagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt
     6241 catcacgcgc tcccacttga agccctcggg gaaggacagc ttcaagtagt cggggatgtc
     6301 ggcggggtgc ttcacgtagg ccttggagcc gtacatgaac tgaggggaca ggatgtccca
     6361 ggcgaagggc agggggccac ccttggtcac cttcagcttg gcggtctggg tgccctcgta
     6421 ggggcggccc tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat
     6481 gtgcaccttg aagcgcatga actccttgat gatggccatg ttatcctcct cgcccttgct
     6541 cacacctgca ggtggttcac gttgaccttg tgggtgatgc tccagtcgtg cacgaagttg
     6601 ttcaagttcc ccagccagtc cttcaacaac gtgtccacct cgccaaccag tggtcttctc
     6661 gtcggaattc gtgacaagtg caagacttag tgcaatgcaa gacaggagtt gcatcctgta
     6721 catggcgcgc ctcccgggtt gtggcaagct tatcatcgtg tttttcaaag gaaaaccacg
     6781 tccccgtggt tcggggggcc tagacgtttt tttaacctcg actaaacaca tgtaaagcat
     6841 gtgcaccgag gccccagatc agatcccata caatggggta ccttctgggc atccttcagc
     6901 cccttgttga atacgcttga ggagagccat ttgactcttt ccacaactat ccaactcaca
     6961 acgtggcact ggggttgtgc cgcctttgca ggtgtatctt atacacgtgg cttttggccg
     7021 cagaggcacc tgtcgccagg tggggggttc cgctgcctgc aaagggtcgc tacagacgtt
     7081 gtttgtcttc aagaagcttc cagaggaact gcttccttca cgacat
//
