LOCUS       sequence-170513-        7031 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 7031)
  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 7031)
  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-DFCP1
FEATURES             Location/Qualifiers
     source          1..7031
                     /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    3819..3940
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3856..3875)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3910..3929
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3947..4402)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4084..4105)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4296..4315
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4429..4533
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4535..4892
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4535..4726
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4558..4578)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4743..4878
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4927..5721
                     /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             4927..5721
                     /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(4981..5000)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5591..5610
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5909..5928)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5953..6000
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6329..6917
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6818..6837
                     /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 agtgcccaga cttccccagc agagaagggc
     1381 ctgaatccgg ggctgatgtg ccaggaaagt tacgcttgca gcgggactga tgaagctatc
     1441 tttgagtgtg atgagtgctg cagtctgcag tgtctccgct gcgaggagga gctccatcgg
     1501 caggagcgcc tgagaaacca tgagcggata agactcaaac ctggccatgt cccttactgt
     1561 gacctctgca agggtctcag tgggcattta ccaggtgtta ggcagagggc aatagtgagg
     1621 tgccagacct gcaaaattaa cttgtgcctg gagtgccaga agaggactca ttctgggggt
     1681 aacaaaagga gacaccctgt tactgtgtac aatgtcagta atctccagga gtcactggag
     1741 gcagaagaga tggatgagga gaccaagagg aagaagatga ctgagaaggt tgtgagtttc
     1801 ctcctagtag acgaaaatga agaaattcag gtaacaaatg aagaagactt tattagaaaa
     1861 ttggactgca aacctgatca gcatctgaaa gtggtttcca tttttggaaa tactggtgat
     1921 ggaaagtctc atactctcaa ccacactttc ttttatggtc gtgaagtctt taaaacctcc
     1981 ccgacccagg agtcctgcac tgtgggagtg tgggcagcct atgacccagt tcacaaagta
     2041 gcagtgatcg atacggaagg gctcctgggg gccaccgtga atctaagcca gagaacacgg
     2101 ctgctgctta aggtcctggc catctcagac ctcgtcatct atcgaactca tgcagaccgg
     2161 ctgcataacg acctcttcaa attccttggg gatgcctcag aagcttatct gaagcacttc
     2221 accaaggagc tcaaggccac cactgctcgc tgtggcctgg atgtcccttt atccacactg
     2281 ggccctgcag ttatcatctt ccatgagacc gtgcacaccc agctactggg ctctgatcat
     2341 ccctcagagg tgccagagaa gctcatccag gaccggttcc ggaagctggg ccgtttccct
     2401 gaagccttta gttccattca ctacaaggga acgaggactt acaaccctcc cacggacttt
     2461 tctgggcttc ggcgtgcttt ggagcagcta ctagagaata acaccacccg ttctccccgg
     2521 cacccgggag tcatcttcaa agccctgaag gcactaagtg accgcttcag cggtgagatc
     2581 cccgatgacc agatggcgca cagctccttt tttccagatg agtatttcac ctgctcctcc
     2641 ttgtgcctca gctgtggggt tggatgtaag aaaagcatga atcatgggaa ggaaggagtg
     2701 cctcatgaag ccaagagccg ctgcagatac tcccaccagt atgacaaccg agtgtatacc
     2761 tgcaaggcct gctatgagag aggcgaggaa gtcagtgtag tgcccaaaac atctgcttcc
     2821 actgactccc cctggatggg tctcgcaaaa tatgcctggt ctgggtatgt gatcgaatgt
     2881 cctaactgtg gcgtggtcta tcgtagtcgg cagtactggt ttggaaacca agatcctgtg
     2941 gatacggtgg tgcggacaga gattgtgcat gtgtggcctg gaactgatgg gtttctgaag
     3001 gacaacaaca atgctgccca gcgcctgttg gacgggatga acttcatggc tcagtcggtg
     3061 tccgagctta gccttggacc caccaaggct gtgacttcct ggctgacaga ccagatcgcc
     3121 cctgcctact ggaggcccaa ctcccagatt ctgagctgca acaagtgtgc gacgtccttt
     3181 aaagataacg acactaagca tcactgccga gcctgtgggg agggcttctg tgacagctgt
     3241 tcatcaaaga ctcggccagt gcctgagcgg ggctggggcc ctgcgccagt gcgggtctgt
     3301 gacaactgct acgaagccag gaacgtccag ttagctgtta ccgaggcaca agtggacgat
     3361 gaaggtggaa cgctcattgc tcggaaggtg ggcgaggccg tgcagaacac tctgggagcc
     3421 gtggtgacag ccattgacat accactaggt ctggtaaagg acgcggccag gcctgcgtac
     3481 tgggtgcctg accacgaaat cctccactgc cacaactgcc ggaaggagtt cagcatcaag
     3541 ctctccaagc accactgccg ggcctgcgga cagggcttct gtgatgagtg ctcccatgac
     3601 cgccgggctg ttccttctcg tggctgggac catcccgtcc gagtctgctt caactgcaat
     3661 aaaaagcccg gtgaccttta agcggccgca gatccaccgg atctagataa ctgatcataa
     3721 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3781 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3841 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3901 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     3961 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4021 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4081 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4141 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     4201 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     4261 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4321 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4381 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4441 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4501 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     4561 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4621 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4681 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4741 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4801 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     4861 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     4921 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4981 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5041 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5101 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5161 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5221 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5281 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5341 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5401 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     5461 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5521 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5581 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5641 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5701 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5761 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5821 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5881 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     5941 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     6001 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     6061 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     6121 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     6181 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6241 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6301 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6361 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6421 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6481 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6541 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6601 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6661 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6721 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6781 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6841 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6901 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6961 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7021 ccgccatgca t
//
