LOCUS       sequence-384085-        6553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of siRNA-resistant mutant ABHD16A tagged with 
            Halo tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6553)
  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 6553)
  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-Halo
FEATURES             Location/Qualifiers
     source          1..6553
                     /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"
     CDS             2326..3216
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     polyA_signal    3341..3462
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3378..3397)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3432..3451
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3469..3924)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3606..3627)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3818..3837
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3951..4055
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4057..4414
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4057..4248
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4080..4100)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4265..4400
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4449..5243
                     /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             4449..5243
                     /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(4503..4522)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5113..5132
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5431..5450)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5475..5522
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5851..6439
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6340..6359
                     /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 cggtcatggc agaaatcggt
     2341 actggctttc cattcgaccc ccattatgtg gaagtcctgg gcgagcgcat gcactacgtc
     2401 gatgttggtc cgcgcgatgg cacccctgtg ctgttcctgc acggtaaccc gacctcctcc
     2461 tacgtgtggc gcaacatcat cccgcatgtt gcaccgaccc atcgctgcat tgctccagac
     2521 ctgatcggta tgggcaaatc cgacaaacca gacctgggtt atttcttcga cgaccacgtc
     2581 cgcttcatgg atgccttcat cgaagccctg ggtctggaag aggtcgtcct ggtcattcac
     2641 gactggggct ccgctctggg tttccactgg gccaagcgca atccagagcg cgtcaaaggt
     2701 attgcattta tggagttcat ccgccctatc ccgacctggg acgaatggcc agaatttgcc
     2761 cgcgagacct tccaggcctt ccgcaccacc gacgtcggcc gcaagctgat catcgatcag
     2821 aacgttttta tcgagggtac gctgccgatg ggtgtcgtcc gcccgctgac tgaagtcgag
     2881 atggaccatt accgcgagcc gttcctgaat cctgttgacc gcgagccact gtggcgcttc
     2941 ccaaacgagc tgccaatcgc cggtgagcca gcgaacatcg tcgcgctggt cgaagaatac
     3001 atggactggc tgcaccagtc ccctgtcccg aagctgctgt tctggggcac cccaggcgtt
     3061 ctgatcccac cggccgaagc cgctcgcctg gccaaaagcc tgcctaactg caaggctgtg
     3121 gacatcggcc cgggtctgaa tctgctgcaa gaagacaacc cggacctgat cggcagcgag
     3181 atcgcgcgct ggctgtcgac gctcgagatt tccggctgag cggccgcgac tctagatcat
     3241 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3301 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3361 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3421 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     3481 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3541 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3601 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3661 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3721 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3781 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3841 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3901 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3961 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4021 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     4081 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4141 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4201 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4261 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4321 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4381 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     4441 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4501 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4561 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4621 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4681 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4741 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4801 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4861 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4921 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     4981 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5041 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5101 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5161 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5221 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5281 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5341 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5401 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5461 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5521 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5581 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     5641 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     5701 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5761 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5821 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5881 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5941 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6001 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6061 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6121 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6181 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6241 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6301 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6361 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6421 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6481 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6541 taccgccatg cat
//
