LOCUS       sequence-384021-        6226 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of outer mitochondrial membrane localized 
            ABHD16A tagged with mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6226)
  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 6226)
  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: Tom70NTD-ABHD16Acyto-mNG
FEATURES             Location/Qualifiers
     source          1..6226
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..229
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     180..200
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1837..1846
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1843..2553
                     /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      2098..2107
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2675..2796
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2712..2731)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2766..2785
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2803..3258)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2940..2961)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3152..3171
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3285..3389
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3391..3748
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3391..3582
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3414..3434)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3599..3734
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3783..4577
                     /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             3783..4577
                     /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(3837..3856)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4447..4466
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4765..4784)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4809..4856
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5185..5773
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5674..5693
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(5948..6226,1..25)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
       61 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      121 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      181 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
      241 ccgtcagatc cgctagcgct accggactca gatctcgagc tcaagcttat ggccgcctct
      301 aaacctgtgg aggcagcggt ggtcgcagcc gctgtaccga gctccgggag tggggtgggc
      361 ggcggcggga ctgcgggccc gggcacgggg gggctgccgc gatggcagct ggctctggcg
      421 gtcggggcac ccctgctgct gggcgcgggt gccatatacc tgtggagtcg gcagcaacgg
      481 cgccggggcc gctggaccaa cccccagtac cggcagttca tcaccatctt ggaagcaaca
      541 catcggaacc agtcttcaga aaacaagagg cagcttgcca actacaactt tgacttccgg
      601 agctggccag tcgacttcca ctgggaagaa cccagcagcc ggaaggagtc tcgagggggc
      661 ccttcccgcc ggggtgtggc cctgcttcgc ccagagcccc tgcaccgggg gacagcagac
      721 accctcctca accgggttaa gaagctgcct tgtcagatca ccagctacct ggtggcgcac
      781 accctagggc gccggatgct gtatccaggc tctgtgtacc tgctgcagaa ggccctcatg
      841 cctgtgctgc tgcagggcca ggcccgactg gtggaagagt gtaatgggcg ccgggcaaag
      901 ctgctggcct gtgatggcaa tgagattgac accatgtttg tggaccggcg ggggacagct
      961 gagccccagg gacagaagct ggtgatctgc tgtgagggga atgctgggtt ttatgaggtg
     1021 ggctgcgtct ccacgcccct ggaagctgga tattcagtcc tgggctggaa tcatccaggc
     1081 tttgctggaa gcacgggggt gccattcccg cagaatgagg ctaatgccat ggatgtggtg
     1141 gtccagtttg ccatccaccg cctgggcttc cagccccagg acatcatcat ctacgcctgg
     1201 tccatcggcg gcttcactgc cacgtgggca gccatgtcct acccagatgt tagtgccatg
     1261 atcctggatg cctcctttga tgacctggtg cccttggcct tgaaggtcat gccagacagc
     1321 tggaggggcc tggtgaccag gaccgtgagg cagcatctca atctaaacaa cgcggagcag
     1381 ctgtgcagat accagggtcc tgtactgctg atccggagaa ccaaggatga gatcatcacc
     1441 accacggttc ctgaggacat catgtccaac cgaggcaatg acctcctgct gaagctcctg
     1501 cagcatcggt atccccgggt gatggcagag gagggtcttc gagtggtgag gcagtggttg
     1561 gaggcctcct cacagctgga ggaagcctca atttatagcc gatgggaggt ggaagaggac
     1621 tggtgtctgt ctgtcctccg ctcctaccag gcagaacacg ggcctgactt cccctggagc
     1681 gtgggggagg acatgagtgc agatggacgg cggcagctgg ctttgtttct ggctcggaag
     1741 catctgcaca actttgaggc cactcactgc accccactcc cagcccagaa cttccagatg
     1801 ccctggcacc tcccgcgggc ccgggatcca ccggtcgcca ccatggtgag caagggcgag
     1861 gaggataaca tggcctctct cccagcgaca catgagttac acatctttgg ctccatcaac
     1921 ggtgtggact ttgacatggt gggtcagggc accggcaatc caaatgatgg ttatgaggag
     1981 ttaaacctga agtccaccaa gggtgacctc cagttctccc cctggattct ggtccctcat
     2041 atcgggtatg gcttccatca gtacctgccc taccctgacg ggatgtcgcc tttccaggcc
     2101 gccatggtag atggctccgg ctaccaagtc catcgcacaa tgcagtttga agatggtgcc
     2161 tcccttactg ttaactaccg ctacacctac gagggaagcc acatcaaagg agaggcccag
     2221 gtgaagggga ctggtttccc tgctgacggt cctgtgatga ccaactcgct gaccgctgcg
     2281 gactggtgca ggtcgaagaa gacttacccc aacgacaaaa ccatcatcag tacctttaag
     2341 tggagttaca ccactggaaa tggcaagcgc taccggagca ctgcgcggac cacctacacc
     2401 tttgccaagc caatggcggc taactatctg aagaaccagc cgatgtacgt gttccgtaag
     2461 acggagctca agcactccaa gaccgagctc aacttcaagg agtggcaaaa ggcctttacc
     2521 gatgtgatgg gcatggacga gctgtacaag taagcggccg cgactctaga tcataatcag
     2581 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2641 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2701 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2761 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     2821 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     2881 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     2941 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3001 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3061 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3121 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3181 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3241 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3301 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3361 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     3421 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3481 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3541 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3601 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3661 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3721 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     3781 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3841 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3901 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3961 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4021 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4081 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4141 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4201 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4261 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     4321 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4381 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4441 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4501 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4561 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4621 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4681 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4741 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     4801 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     4861 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4921 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     4981 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5041 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5101 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5161 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5221 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5281 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5341 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5401 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5461 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5521 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5581 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5641 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5701 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5761 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5821 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5881 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     5941 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     6001 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     6061 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     6121 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     6181 cccagtacat gaccttatgg gactttccta cttggcagta catcta
//
