LOCUS       sequence-384019-        6730 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of ER localized ABHD16A tagged with mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6730)
  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 6730)
  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: Stim1NTD-ABHD16Acyto-mNG
FEATURES             Location/Qualifiers
     source          1..6730
                     /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      2680..2689
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2686..3396
                     /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      2941..2950
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3518..3639
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3555..3574)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3609..3628
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3646..4101)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3783..3804)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3995..4014
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4128..4232
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4234..4591
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4234..4425
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4257..4277)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4442..4577
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4626..5420
                     /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             4626..5420
                     /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(4680..4699)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5290..5309
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5608..5627)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5652..5699
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6028..6616
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6517..6536
                     /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 gatgtatgcg tccgtcttgc cctgtggctc
      661 ctctggggac tcctcctgca ccagggccag agcctcagcc atagtcacag tgagaaggcg
      721 acaggaacca gctcgggggc caactctgag gagtccactg cagcagagtt ttgccgaatt
      781 gacaagcccc tgtgtcacag tgaggatgag aaactcagct tcgaggcagt ccgtaacatc
      841 cacaaactga tggacgatga tgccaatggt gatgtggatg tggaagaaag tgatgagttc
      901 ctgagggaag acctcaatta ccatgaccca acagtgaaac acagcacctt ccatggtgag
      961 gataagctca tcagcgtgga ggacctgtgg aaggcatgga agtcatcaga agtatacaat
     1021 tggaccgtgg atgaggtggt acagtggctg atcacatatg tggagctgcc tcagtatgag
     1081 gagaccttcc ggaagctgca gctcagtggc catgccatgc caaggctggc tgtcaccaac
     1141 accaccatga cagggactgt gctgaagatg acagaccgga gtcatcggca gaagctgcag
     1201 ctgaaggctc tggatacagt gctctttggg cctcctctct tgactcgcca taatcacctc
     1261 aaggacttca tgctggtggt gtctatcgtt attggtgtgg gcggctgctg gtttgcctat
     1321 atccagaacg gccgctggac caacccccag taccggcagt tcatcaccat cttggaagca
     1381 acacatcgga accagtcttc agaaaacaag aggcagcttg ccaactacaa ctttgacttc
     1441 cggagctggc cagtcgactt ccactgggaa gaacccagca gccggaagga gtctcgaggg
     1501 ggcccttccc gccggggtgt ggccctgctt cgcccagagc ccctgcaccg ggggacagca
     1561 gacaccctcc tcaaccgggt taagaagctg ccttgtcaga tcaccagcta cctggtggcg
     1621 cacaccctag ggcgccggat gctgtatcca ggctctgtgt acctgctgca gaaggccctc
     1681 atgcctgtgc tgctgcaggg ccaggcccga ctggtggaag agtgtaatgg gcgccgggca
     1741 aagctgctgg cctgtgatgg caatgagatt gacaccatgt ttgtggaccg gcgggggaca
     1801 gctgagcccc agggacagaa gctggtgatc tgctgtgagg ggaatgctgg gttttatgag
     1861 gtgggctgcg tctccacgcc cctggaagct ggatattcag tcctgggctg gaatcatcca
     1921 ggctttgctg gaagcacggg ggtgccattc ccgcagaatg aggctaatgc catggatgtg
     1981 gtggtccagt ttgccatcca ccgcctgggc ttccagcccc aggacatcat catctacgcc
     2041 tggtccatcg gcggcttcac tgccacgtgg gcagccatgt cctacccaga tgttagtgcc
     2101 atgatcctgg atgcctcctt tgatgacctg gtgcccttgg ccttgaaggt catgccagac
     2161 agctggaggg gcctggtgac caggaccgtg aggcagcatc tcaatctaaa caacgcggag
     2221 cagctgtgca gataccaggg tcctgtactg ctgatccgga gaaccaagga tgagatcatc
     2281 accaccacgg ttcctgagga catcatgtcc aaccgaggca atgacctcct gctgaagctc
     2341 ctgcagcatc ggtatccccg ggtgatggca gaggagggtc ttcgagtggt gaggcagtgg
     2401 ttggaggcct cctcacagct ggaggaagcc tcaatttata gccgatggga ggtggaagag
     2461 gactggtgtc tgtctgtcct ccgctcctac caggcagaac acgggcctga cttcccctgg
     2521 agcgtggggg aggacatgag tgcagatgga cggcggcagc tggctttgtt tctggctcgg
     2581 aagcatctgc acaactttga ggccactcac tgcaccccac tcccagccca gaacttccag
     2641 atgccctggc acctcccgcg ggcccgggat ccaccggtcg ccaccatggt gagcaagggc
     2701 gaggaggata acatggcctc tctcccagcg acacatgagt tacacatctt tggctccatc
     2761 aacggtgtgg actttgacat ggtgggtcag ggcaccggca atccaaatga tggttatgag
     2821 gagttaaacc tgaagtccac caagggtgac ctccagttct ccccctggat tctggtccct
     2881 catatcgggt atggcttcca tcagtacctg ccctaccctg acgggatgtc gcctttccag
     2941 gccgccatgg tagatggctc cggctaccaa gtccatcgca caatgcagtt tgaagatggt
     3001 gcctccctta ctgttaacta ccgctacacc tacgagggaa gccacatcaa aggagaggcc
     3061 caggtgaagg ggactggttt ccctgctgac ggtcctgtga tgaccaactc gctgaccgct
     3121 gcggactggt gcaggtcgaa gaagacttac cccaacgaca aaaccatcat cagtaccttt
     3181 aagtggagtt acaccactgg aaatggcaag cgctaccgga gcactgcgcg gaccacctac
     3241 acctttgcca agccaatggc ggctaactat ctgaagaacc agccgatgta cgtgttccgt
     3301 aagacggagc tcaagcactc caagaccgag ctcaacttca aggagtggca aaaggccttt
     3361 accgatgtga tgggcatgga cgagctgtac aagtaagcgg ccgcgactct agatcataat
     3421 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     3481 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3541 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3601 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     3661 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3721 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3781 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3841 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3901 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3961 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     4021 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     4081 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4141 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4201 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     4261 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4321 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4381 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4441 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4501 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4561 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     4621 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4681 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4741 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4801 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4861 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4921 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4981 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5041 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5101 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     5161 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5221 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5281 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5341 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5401 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     5461 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5521 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5581 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     5641 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     5701 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     5761 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     5821 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     5881 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5941 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6001 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6061 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6121 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6181 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6241 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6301 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6361 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6421 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6481 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6541 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6601 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6661 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6721 cgccatgcat
//
