LOCUS       sequence-322489-        6144 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6144)
  AUTHORS   Maejima I, Takahashi A, Omori H, Kimura T, Takabatake Y, Saitoh T, 
            Yamamoto A, Hamasaki M, Noda T, Isaka Y, Yoshimori T
  TITLE     Autophagy sequesters damaged lysosomes to control lysosomal 
            biogenesis and kidney injury.
  JOURNAL   EMBO J. 2013 Aug 28;32(17):2336-47. doi: 10.1038/emboj.2013.171. 
            Epub 2013 Aug 6.
  PUBMED    23921551
REFERENCE   2  (bases 1 to 6144)
  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: 
            "10.1038/emboj.2013.171"; journalName: "EMBO J"; date: "2013-08-28- 
            28"; volume: "32"; issue: "17"; pages: "2336-47"
COMMENT     Sequence Label: ptf-Galectin3
FEATURES             Location/Qualifiers
     source          1..6144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      605..614
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             611..1285
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             611..1285
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(741..759)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1009..1028
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1203..1226
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     regulatory      1298..1307
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1304..1306,1307..1309,1310..2020)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1304..1306,1307..1309,1310..2020)
                     /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(1349..1370)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1610..1629)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1957..1978
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2939..3060
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2976..2995)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3030..3049
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3067..3522)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3204..3225)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3416..3435
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3549..3653
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3655..4012
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3655..3846
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3678..3698)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3863..3998
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4047..4841
                     /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             4047..4841
                     /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(4101..4120)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4711..4730
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5029..5048)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5073..5120
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5449..6037
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5938..5957
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcggccacc atggcctcct ccgaggacgt catcaaggag ttcatgcgct tcaaggtgcg
      661 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
      721 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc
      781 ctgggacatc ctgtcccctc agttccagta cggctccaag gcctacgtga agcaccccgc
      841 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
      901 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
      961 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     1021 gaagaagacc atgggctggg aggcctccac cgagcggatg taccccgagg acggcgccct
     1081 gaagggcgag atcaagatga ggctgaagct gaaggacggc ggccactacg acgccgaggt
     1141 caagaccacc tacatggcca agaagcccgt gcagctgccc ggcgcctaca agaccgacat
     1201 caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgagcgcgc
     1261 cgagggccgc cactccaccg gcgccggtgg accggtcgcc accatggtga gcaagggcga
     1321 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1381 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1441 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     1501 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1561 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1621 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1681 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1741 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     1801 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     1861 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     1921 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1981 cgccgccggg atcactctcg gcatggacga gctgtacaag tccggactca gatctatggc
     2041 agacaatttt tcgctccatg atgcgttatc tgggtctgga aacccaaacc ctcaaggatg
     2101 gcctggcgca tgggggaacc agcctgctgg ggcagggggc tacccagggg cttcctatcc
     2161 tggggcctac cccgggcagg cacccccagg ggcttatcct ggacaggcac ctccaggcgc
     2221 ctaccctgga gcacctggag cttatcccgg agcacctgca cctggagtct acccagggcc
     2281 acccagcggc cctggggcct acccatcttc tggacagcca agtgcccccg gagcctaccc
     2341 tgccactggc ccctatggcg cccctgctgg gccactgatt gtgccttata acctgccttt
     2401 gcctggggga gtggtgcctc gcatgctgat aacaattctg ggcacggtga agcccaatgc
     2461 aaacagaatt gctttagatt tccaaagagg gaatgatgtt gccttccact ttaacccacg
     2521 cttcaatgag aacaacagga gagtcattgt ttgcaataca aagctggata ataactgggg
     2581 aagggaagaa agacagtcgg ttttcccatt tgaaagtggg aaaccattca aaatacaagt
     2641 actggttgaa cctgaccact tcaaggttgc agtgaatgat gctcacttgt tgcagtacaa
     2701 tcatcgggtt aaaaaactca atgaaatcag caaactggga atttctggtg acatagacct
     2761 caccagtgct tcatatacca tgatataagt cgacggtacc gcgggcccgg gatccaccgg
     2821 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     2881 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     2941 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3001 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3061 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3121 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3181 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3241 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3301 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3361 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3421 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     3481 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     3541 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3601 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     3661 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3721 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3781 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3841 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3901 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3961 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     4021 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4081 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4141 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4201 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     4261 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4321 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4381 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4441 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4501 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4561 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4621 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4681 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4741 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4801 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4861 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4921 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4981 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     5041 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     5101 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     5161 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     5221 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     5281 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5341 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5401 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5461 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5521 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5581 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5641 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5701 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5761 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5821 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5881 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5941 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6001 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6061 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6121 ttctgtggat aaccgtatta ccgc
//
