LOCUS       sequence-258500-        4968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  M11-92 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4968)
  AUTHORS   Chang CL, Weigel AV, Ioannou MS, Pasolli HA, Xu CS, Peale DR, 
            Shtengel G, Freeman M, Hess HF, Blackstone C, Lippincott-Schwartz J
  TITLE     Spastin tethers lipid droplets to peroxisomes and directs fatty acid
            trafficking through ESCRT-III.
  JOURNAL   J Cell Biol. 2019 Aug 5;218(8):2583-2599. doi: 
            10.1083/jcb.201902061. Epub 2019 Jun 21.
  PUBMED    31227594
REFERENCE   2  (bases 1 to 4968)
  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.201902061"; journalName: "J Cell Biol"; date: 
            "2019-08-5- 5"; volume: "218"; issue: "8"; pages: "2583-2599"
COMMENT     Sequence Label: mApple-M11-92
FEATURES             Location/Qualifiers
     source          1..4968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        320..523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     474..494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      562..571
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             568..1275
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK
                     SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     981..1000
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1711..1832
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1748..1767)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1802..1821
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1839..2294)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1976..1997)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2188..2207
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2321..2425
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2427..2784
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2427..2618
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2450..2470)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2635..2770
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2819..3613
                     /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             2819..3613
                     /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(2873..2892)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3483..3502
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3801..3820)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3845..3892
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4221..4809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4710..4729
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
       61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      541 gatccgctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagaa taacatggcc
      601 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
      661 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg cctttcagac cgctaagctg
      721 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
      781 tacggctcca aggtctacat taagcaccca gccgacatcc ccgactactt caagctgtcc
      841 ttccccgagg gcttcaggtg ggagcgcgtg atgaacttcg aggacggcgg cattattcac
      901 gttaaccagg actcctccct gcaggacggc gtgttcatct acaaggtgaa gctgcgcggc
      961 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     1021 gaggagcgga tgtaccccga ggacggcgcc ctgaagagcg agatcaagaa gaggctgaag
     1081 ctgaaggacg gcggccacta cgccgccgag gtcaagacca cctacaaggc caagaagccc
     1141 gtgcagctgc ccggcgccta catcgtcgac atcaagttgg acatcgtgtc ccacaacgag
     1201 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     1261 gacgagctgt acaagtccgg actcagatct cgagctcaag cttcgaattc tccgggtgga
     1321 cgagggaaga agaaaggctc cggcggcgcc agcaacccgg tgcctcccag gcctccgccc
     1381 ccttgcctgg cccccgcccc tcccgccgcc gggccggccc ctccgcccga gtcgccgcat
     1441 aagcggaacc tgtactattt ctcctacccg ctgtttgtag gcttcgcgct gctgcgtttg
     1501 gtcgccttcc acctggggct cctcttcgtg tggctctgcc agcgcttctc ccgcgccctc
     1561 atggcagcca agaggagcta aggatccacc ggatctagat aactgatcat aatcagccat
     1621 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     1681 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     1741 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     1801 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     1861 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     1921 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     1981 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     2041 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     2101 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     2161 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     2221 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     2281 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2341 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2401 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     2461 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2521 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2581 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     2641 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     2701 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     2761 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     2821 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     2881 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     2941 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3001 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3061 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3121 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     3181 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     3241 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     3301 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     3361 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     3421 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     3481 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     3541 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     3601 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     3661 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     3721 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     3781 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     3841 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     3901 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     3961 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     4021 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     4081 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4141 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4201 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4261 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4321 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4381 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4441 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4501 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4561 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4621 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4681 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4741 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4801 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4861 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     4921 cattagttat taatagtaat caattacggg gtcattagtt catagccc
//
