LOCUS       sequence-402545-        6733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RUSH cargo: Str-KDEL_IRES_SBP-EGFP-CD-MPR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6733)
  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 6733)
  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-GFP-RUSH
FEATURES             Location/Qualifiers
     source          1..6733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(250..269)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          252..481
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     complement(1245..1264)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1246..1264)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1309..1512)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1338..1358)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1513..1892
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2064..2083
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2158..2262
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2263..2331,2332..3123)
                     /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(2481..2500)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3294..3882
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3783..3802
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4036..4053
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4170..4191
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4206..4211,4212..4229,4230..4236)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4244..4260
                     /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     4249..4271
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4268..4284
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4268..4284
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4386..4507
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4451..4470
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5417..6127,6128..6130,6131..6133))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(5417..6127,6128..6130,6131..6133))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5459..5480)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5808..5827
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6067..6088
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(6143..6253)
                     /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(6254..6313)
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     misc_feature    join(6327..6335,6336..6338,6339..6733,1..180)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6500..6519)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     join(6729..6733,1..13)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
ORIGIN
        1 cctttggcga gaggggaaag acccctagga atgctcgtca agaagacagg gccaggtttc
       61 cgggccctca cattgccaaa agacggcaat atggtggaaa ataacatata gacaaacgca
      121 caccggcctt attccaagcg gcttcggcca gtaacgttag ggggggggga gggagagggg
      181 gggggggaga ggggcggaat tggccgccct agatgcatgc tcgacctgca gttggacctg
      241 ggagtggaca cctgtggaga gaaaggcaaa gtggatgtca ttgtcactca agtgtatggc
      301 cagatctcaa gcctgccaca cctcaagctt gacaacaaaa agattgtctt ttctgaccag
      361 atggacgcgg ccaccctcaa aggcatcacc gcgggccagg tgaatatcaa atcctcctcg
      421 tttttggaaa ctgacaatct tagcgcagaa gtcatgcccg cttttgagag ggagtactca
      481 ccccaacagc tggccctcgc agacagcgaa ttaattccag cacactggat cagttatcta
      541 tgcggccgct cacagttcat ctttcggtcc gagctgctgg acggcatcca gaggattgcc
      601 gttgttgact ccggcttttt tggcggcatc gattgaagcg gcggagggtt tcactttggt
      661 gaatgtgtca tgtcccacca gggtggattt ccaagcattt gcctcggttg tgccagatgt
      721 gagcagccac tgtgtattga ttctagcctc agcgcctccc acgtactgtc cactccatgt
      781 tgtggcagaa tgagcattcc ggtagttgtt tttccaagcg acagtccatc ccagggcagt
      841 cccagatccg tcggtggcag gggcactatc gtatcggcca gtgaggacgt accgtgattc
      901 ggcgtttccc acagcagatt cgtatgttcc ggtgagggct ccatcggctc cagcagtcac
      961 aataaaggtg ctcccgagct gattgtacca tgttccagta atgccagcct cggcagcgga
     1021 cacctgagct tttgagtctt tgctagggtc gtttaaaccg gcgcccgagc tggttcctgt
     1081 cgccttctca ctgtgactat ggctgaggct ctggccctgg tgcaggagga gtccccagag
     1141 gagccacagg gcaagacgga cgcatacatc cataccggtt aacaggcctt aagcgctagc
     1201 taggccgcag atatcgatcc gagctcggta ccaagcttgg gtctccctat agtgagtcgt
     1261 attaatttcg ataagccagt aagcagtggg ttctctagtt agccagagag ctctgcttat
     1321 atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg
     1381 acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg
     1441 gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca
     1501 aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
     1561 gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
     1621 aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
     1681 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
     1741 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
     1801 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
     1861 ttactattaa taactagtca ataatcaatg tcaacgcgta tatctggccc gtacatcgcg
     1921 aagcagcgca aaacgcctaa ccctaagcag attcttcatg caattgtcgg tcaagccttg
     1981 ccttgttgta gcttaaattt tgctcgcgca ctactcagcg acctccaaca cacaagcagg
     2041 gagcagatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatag
     2101 gggatcggga gatctcccga tccgtcgacg tcaggtggca cttttcgggg aaatgtgcgc
     2161 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2221 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     2281 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     2341 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     2401 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     2461 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     2521 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     2581 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     2641 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     2701 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2761 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     2821 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2881 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2941 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3001 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3061 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3121 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3181 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3241 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3301 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3361 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3421 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     3481 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3541 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3601 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3661 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3721 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3781 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3841 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3901 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3961 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4021 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     4081 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     4141 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     4201 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     4261 tttcacacag gaaacagcta tgaccatgat tacgccaagc tctagctaga ggtcgacggt
     4321 atacagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcactcga
     4381 cgcgttaaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     4441 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     4501 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     4561 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcagttatc
     4621 tagagttaat taattggccc tcgaggccct acattggtaa taaatggtca tccctttctt
     4681 ctgactcctc ccccagctgg tcatccccca caccacgata tgctgcaggc acatttcgag
     4741 gtttagaacg gcagacaaag tcacagccat ctgctaccag gttgccaaga tcctgccaga
     4801 aggctaagtg gggaaactgc tccattcctt tggctcccac taccagtcgc tggtatagga
     4861 accccccaac aacataaaca gcaaccagtg atgcaaacgt gacaagtaag atggaaccca
     4921 cactgaggtg ggagatctct ggtgaacagg ccaggctgct atccatctca aagaggtaga
     4981 aacaatcttg gactttgcca cgctcctcag acacagggtt aaaattgtcc gctagggtgt
     5041 gtcgattgca ggagatcatc accactgcac gacgctgctc cttgccacag tggttgtcat
     5101 attcatcacc ccctttatag atcagcatga tccaattact tccgttgaag atgtgagtct
     5161 cgttgagtct ccctaccact gtctccttcc cattactttt gttgatttgc accaggcctg
     5221 ccccagaagt gtggttgcca gcttcccggc acaccctgaa gatgtagatg tatgtgtctg
     5281 aaccctggcc cacagtgctc tcaaagcttt tattaaacag tggtttcagc ctcttcacta
     5341 gagccaactc tttctctgac tctctaccct tttctcctac caagtcgcaa gttttttctt
     5401 ctgtgtctgg ccggcccttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     5461 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact
     5521 gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     5581 gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt
     5641 tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt
     5701 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     5761 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
     5821 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     5881 agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca
     5941 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca
     6001 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
     6061 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     6121 ccttgctcac catacctgca ggtggttcac gttgaccttg tgggtgatgc tccagtcgtg
     6181 cacgaagttg ttcaagttcc ccagccagtc cttcaacaac gtgtccacct cgccaaccag
     6241 tggtcttctc gtcggaattc gtgacaagtg caagacttag tgcaatgcaa gacaggagtt
     6301 gcatcctgta catggcgcgc ctcccgggtt gtggcaagct tatcatcgtg tttttcaaag
     6361 gaaaaccacg tccccgtggt tcggggggcc tagacgtttt tttaacctcg actaaacaca
     6421 tgtaaagcat gtgcaccgag gccccagatc agatcccata caatggggta ccttctgggc
     6481 atccttcagc cccttgttga atacgcttga ggagagccat ttgactcttt ccacaactat
     6541 ccaactcaca acgtggcact ggggttgtgc cgcctttgca ggtgtatctt atacacgtgg
     6601 cttttggccg cagaggcacc tgtcgccagg tggggggttc cgctgcctgc aaagggtcgc
     6661 tacagacgtt gtttgtcttc aagaagcttc cagaggaact gcttccttca cgacattcaa
     6721 cagaccttgc att
//
