LOCUS       sequence-170512-        6797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal mCherry-tagged protein expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6797)
  AUTHORS   Kim YM, Jung CH, Seo M, Kim EK, Park JM, Bae SS, Kim DH
  TITLE     mTORC1 phosphorylates UVRAG to negatively regulate autophagosome and
            endosome maturation.
  JOURNAL   Mol Cell. 2015 Jan 22;57(2):207-18. doi: 
            10.1016/j.molcel.2014.11.013. Epub 2014 Dec 18.
  PUBMED    25533187
REFERENCE   2  (bases 1 to 6797)
  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.molcel.2014.11.013"; journalName: "Mol Cell"; date: 
            "2015-01-22- 22"; volume: "57"; issue: "2"; pages: "207-18"
COMMENT     Sequence Label: mCherry-UVRAG S498D
FEATURES             Location/Qualifiers
     source          1..6797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1320
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(758..776)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1026..1045
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3585..3706
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3622..3641)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3676..3695
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3713..4168)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3850..3871)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4062..4081
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4195..4299
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4301..4658
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4301..4492
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4324..4344)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4509..4644
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4693..5487
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4693..5487
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4747..4766)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5357..5376
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5675..5694)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5719..5766
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6095..6683
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6584..6603
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
      661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      721 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      781 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
      841 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      961 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1081 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1141 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1201 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     1261 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1321 tccggactca gatctcgagg gtcgaccatg agcgcctccg cgtcggtcgg gggccaggtc
     1381 ccccagccac ccccgggccc ggccgctgct ctgcctcccg gttctgccgc gcgggccctg
     1441 catgtggagc tgccgtctca gcagcggcgt cttcgacatc ttcggaacat tgctgcccgg
     1501 aacattgtta atagaaatgg ccatcagctc cttgatacct actttacact tcacttgtgt
     1561 agtactgaaa agatatataa agaattttat agaagtgaag tgattaagaa ttccttgaat
     1621 cccacgtggc gaagtctcga ttttggaatt atgccagacc gtcttgatac atctgtgtct
     1681 tgtttcgtgg tgaagatatg gggtggaaag gagaacatct accagctgtt gattgaatgg
     1741 aaagtctgtt tggatgggct gaaatacttg ggtcagcaga ttcatgcccg aaaccaaaat
     1801 gaaataattt ttgggctgaa tgatggatac tatggtgctc catttgaaca taagggttat
     1861 tcaaatgctc agaagactat tcttctgcag gtggatcaga actgtgttcg caattcttac
     1921 gatgtcttct ctttgctacg gcttcataga gcccagtgtg caattaaaca gactcaggta
     1981 actgttcaga aaattggaaa ggaaattgaa gaaaaactaa gactcacatc tacaagcaat
     2041 gaactgaaaa aaaaaagtga atgcctgcag ttaaaaattt tggtgcttca gaatgaactg
     2101 gaacggcaga agaaagcttt gggacgggag gtggcattac tgcataagca acaaattgca
     2161 ttacaagaca aaggaagtgc attttcagct gagcacctca aacttcaact ccagaaggaa
     2221 tccctaaatg agctgaggaa ggagtgcact gcaaaaagag aactcttctt gaagactaat
     2281 gctcagttga caattcgttg caggcagtta ctctctgagc tttcctacat ttaccctatt
     2341 gatttgaatg aacataagga ttactttgta tgcggtgtca agttgcctaa ttctgaggac
     2401 ttccaagcaa aagatgatgg aagcattgct gttgcccttg gttatactgc acatctggtc
     2461 tccatgattt cctttttcct acaagtgccc ctcagatatc ctataattca taaggggtct
     2521 agatcaacaa tcaaagacaa tatcaatgac aaactgacgg aaaaggagag agagtttcca
     2581 ctgtatccaa aaggagggga gaagttgcag tttgattatg gtgtctatct tctgaacaaa
     2641 aatatagcac agctaagata tcaacatgga ctagggactc cagacttgcg gcaaaccctt
     2701 cccaacctga aaaacttcat ggagcatgga ctaatggtca ggtgtgacag acatcacacc
     2761 tccagtgcaa tccctgttcc taagagacaa agctccatat ttgggggtgc agatgtaggc
     2821 ttctctgggg ggatccctga cccagacaaa ggacatcgaa aacgggccag ctctgagaat
     2881 gagagacttc agtacaaaac ccctcctccc agttacaact cagcattagc ccagcctgtg
     2941 accaccgtcc cctccatggg agagaccgag agaaagataa catctctatc ctcctccttg
     3001 gatacctcct tggacttctc caaagaaaac aagaaaaaag gagaggatct agttggcagc
     3061 ttaaacggag gccacgcgaa tgtgcaccct agccaagaac aaggagaagc cctctccggg
     3121 caccgggcca cagtcaatgg cactctccta cccagcgagc aggccgggtc cgccagtgtc
     3181 cagcttccag gcgagttcca cccagtctca gaagctgagc tctgctgtac tgtggagcaa
     3241 gcagaagaaa tcatcgggct ggaagccaca ggtttcgcct caggtgatca gctagaagca
     3301 tttaactgca tcccagtgga cagtgctgtg gcagtagagt gtgacgaaca agttctggga
     3361 gaatttgaag agttctcccg aaggatctat gcactgaatg aaaacgtatc cagcttccgc
     3421 cggccgcgca ggagttccga taagtgagcg gccgcagatc caccggatct agataactga
     3481 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3541 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3601 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3661 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     3721 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3781 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3841 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3901 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3961 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4021 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4081 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4141 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4201 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4261 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4321 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4381 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4441 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4501 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4561 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4621 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4681 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4741 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4801 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4861 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4921 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4981 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5041 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5101 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5161 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5221 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5281 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5341 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5401 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5461 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5521 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5581 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5641 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     5701 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     5761 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     5821 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5881 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5941 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6001 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6061 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6121 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6181 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6241 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6301 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6361 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6421 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6481 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6541 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6601 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6661 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6721 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6781 gtattaccgc catgcat
//
