LOCUS       sequence-384106-        6223 bp    DNA     circular SYN 15-APR-2025
DEFINITION  mammalian expression of siRNA-resistant ABHD16A tagged with SNAP 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6223)
  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 6223)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-SNAP
FEATURES             Location/Qualifiers
     source          1..6223
                     /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             2341..2886
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     polyA_signal    3011..3132
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3048..3067)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3102..3121
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3139..3594)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3276..3297)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3488..3507
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3621..3725
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3727..4084
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3727..3918
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3750..3770)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3935..4070
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4119..4913
                     /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             4119..4913
                     /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(4173..4192)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4783..4802
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5101..5120)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5145..5192
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5521..6109
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6010..6029
                     /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 gcaccggggg
     1201 acagcagaca 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 cgggatccaa gcggtagcgg tggcagcagc
     2341 atggacaaag actgcgaaat gaagcgcacc accctggata gccctctggg caagctggaa
     2401 ctgtctgggt gcgaacaggg cctgcaccgt atcatcttcc tgggcaaagg aacatctgcc
     2461 gccgacgccg tggaagtgcc tgccccagcc gccgtgctgg gcggaccaga gccactgatg
     2521 caggccaccg cctggctcaa cgcctacttt caccagcctg aggccatcga ggagttccct
     2581 gtgccagccc tgcaccaccc agtgttccag caggagagct ttacccgcca ggtgctgtgg
     2641 aaactgctga aagtggtgaa gttcggagag gtcatcagct acagccacct ggccgccctg
     2701 gccggcaatc ccgccgccac cgccgccgtg aaaaccgccc tgagcggaaa tcccgtgccc
     2761 attctgatcc cctgccaccg ggtggtgcag ggcgacctgg acgtgggggg ctacgagggc
     2821 gggctcgccg tgaaagagtg gctgctggcc cacgagggtc acagactggg caagcctggg
     2881 ctgggttgag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     2941 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3001 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3061 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3121 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3181 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3241 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3301 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3361 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3421 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3481 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3541 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     3601 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3661 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3721 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3781 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3841 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3901 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3961 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4021 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4081 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4141 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4201 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4261 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     4321 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4381 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4441 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4501 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4561 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4621 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     4681 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4741 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4801 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4861 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4921 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4981 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5041 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     5101 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5161 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5221 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     5281 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5341 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     5401 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5461 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5521 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5581 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5641 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5701 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5761 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5821 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5881 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5941 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6001 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6061 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6121 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6181 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
