LOCUS       sequence-384087-        6376 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of siRNA-resistant mutant ABHD16A tagged with 
            mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6376)
  AUTHORS   Nguyen TT, Voeltz GK
  TITLE     An ER phospholipid hydrolase drives ER-associated mitochondrial 
            constriction for fission and fusion.
  JOURNAL   Elife. 2022 Nov 30;11:e84279. doi: 10.7554/eLife.84279.
  PUBMED    36448541
REFERENCE   2  (bases 1 to 6376)
  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"; journalName: "Elife."; 
            date: "2022-11-30"; volume: "11:e84279. doi"; pages: " 
            10.7554/eLife.84279"
COMMENT     Sequence Label: siRNA-resistant ABHD16A-H189AD194A-mNG
FEATURES             Location/Qualifiers
     source          1..6376
                     /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      2326..2335
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2332..3042
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      2587..2596
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3164..3285
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3201..3220)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3255..3274
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3292..3747)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3429..3450)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3641..3660
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3774..3878
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3880..4237
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3880..4071
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3903..3923)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4088..4223
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4272..5066
                     /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             4272..5066
                     /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(4326..4345)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4936..4955
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5254..5273)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5298..5345
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5674..6262
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6163..6182
                     /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 ccggactcag atctcgagct caagcttatg gcgaagctgc tgagctgcgt cctaggcccc
      661 cggctctaca aaatctaccg ggagagggac tctgaaaggg ccccggccag cgtccctgag
      721 acgccaacgg cagtcactgc cccccattcc agctcctggg atacgtacta tcagccccgt
      781 gccctggaga aacatgctga cagcatcctg gcactggctt cagtattctg gtccatctct
      841 tattactcct ctcccttcgc cttcttctac ttgtacagga aaggttactt gagtttaagt
      901 aaggtagtac cattttctca ctatgctggg acattgctgc tacttctggc aggtgtggcc
      961 tgcctccgag gcattggccg ctggaccaac ccccagtacc ggcagttcat caccatcttg
     1021 gaagcaacac atcggaacca gtcttcagaa aacaagaggc agcttgccaa ctacaacttt
     1081 gacttccgga gctggccagt cgacttccac tgggaagaac ccagcagccg gaaggagtct
     1141 cgagggggcc cttcccgccg gggtgtggcc ctgcttcgcc cagagcccct ggcccggggg
     1201 acagcaaaca ccctcctcaa ccgggttaag aagctgcctt gtcagatcac cagctacctg
     1261 gtggcgcaca ccctagggcg ccggatgctg tatccaggct ctgtgtacct gctgcagaag
     1321 gccctcatgc ctgtgctgct gcagggccag gcccgactgg tggaagagtg taatgggcgc
     1381 cgggcaaagc tgctggcctg tgatggcaat gagattgaca ccatgtttgt ggaccggcgg
     1441 gggacagctg agccccaggg acagaagctg gtgatctgct gtgaggggaa tgctgggttt
     1501 tatgaggtgg gctgcgtctc cacgcccctg gaagctggat attcagtcct gggctggaat
     1561 catccaggct ttgctggaag cacgggggtg ccattcccgc agaatgaggc taatgccatg
     1621 gatgtggtgg tccagtttgc catccaccgc ctgggcttcc agccccagga catcatcatc
     1681 tacgcctggt ccatcggcgg cttcactgcc acgtgggcag ccatgtccta cccagatgtt
     1741 agtgccatga tcctggatgc ctcctttgat gacctggtgc ccttggcctt gaaggtcatg
     1801 ccagacagct ggaggggcct ggtgaccagg accgtgaggc agcatctcaa tctaaacaac
     1861 gcggagcagc tgtgcagata ccagggtcct gtactgctga tccggagaac caaggatgag
     1921 atcatcacca ccacggttcc tgaggacatc atgtccaacc gaggcaatga cctcctgctg
     1981 aagctcctgc agcatcggta tccccgggtg atggcagagg agggtcttcg agtggtgagg
     2041 cagtggttgg aggcctcctc acagctggag gaagcctcaa tttatagccg atgggaggtg
     2101 gaagaggact ggtgtctgtc tgtcctccgc tcctaccagg cagaacacgg gcctgacttc
     2161 ccctggagcg tgggggagga catgagtgca gatggacggc ggcagctggc tttgtttctg
     2221 gctcggaagc atctgcacaa ctttgaggcc actcactgca ccccactccc agcccagaac
     2281 ttccagatgc cctggcacct cccgcgggcc cgggatccac cggtcgccac catggtgagc
     2341 aagggcgagg aggataacat ggcctctctc ccagcgacac atgagttaca catctttggc
     2401 tccatcaacg gtgtggactt tgacatggtg ggtcagggca ccggcaatcc aaatgatggt
     2461 tatgaggagt taaacctgaa gtccaccaag ggtgacctcc agttctcccc ctggattctg
     2521 gtccctcata tcgggtatgg cttccatcag tacctgccct accctgacgg gatgtcgcct
     2581 ttccaggccg ccatggtaga tggctccggc taccaagtcc atcgcacaat gcagtttgaa
     2641 gatggtgcct cccttactgt taactaccgc tacacctacg agggaagcca catcaaagga
     2701 gaggcccagg tgaaggggac tggtttccct gctgacggtc ctgtgatgac caactcgctg
     2761 accgctgcgg actggtgcag gtcgaagaag acttacccca acgacaaaac catcatcagt
     2821 acctttaagt ggagttacac cactggaaat ggcaagcgct accggagcac tgcgcggacc
     2881 acctacacct ttgccaagcc aatggcggct aactatctga agaaccagcc gatgtacgtg
     2941 ttccgtaaga cggagctcaa gcactccaag accgagctca acttcaagga gtggcaaaag
     3001 gcctttaccg atgtgatggg catggacgag ctgtacaagt aagcggccgc gactctagat
     3061 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     3121 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3181 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3241 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     3301 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3361 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3421 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3481 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3541 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3601 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3661 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3721 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3781 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3841 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     3901 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3961 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4021 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4081 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4141 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4201 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     4261 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4321 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4381 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4441 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4501 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4561 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4621 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4681 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4741 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     4801 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4861 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4921 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4981 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5041 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5101 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5161 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5221 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     5281 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     5341 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     5401 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     5461 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     5521 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5581 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5641 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5701 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5761 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5821 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5881 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5941 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6001 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6061 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6121 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6181 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6241 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6301 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6361 tattaccgcc atgcat
//
