LOCUS       sequence-384091-        6370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of siRNA-resistant ABHD16A tagged with mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6370)
  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 6370)
  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-mCherry
FEATURES             Location/Qualifiers
     source          1..6370
                     /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..3036
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2471..2489)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2739..2758
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3158..3279
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3195..3214)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3249..3268
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3286..3741)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3423..3444)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3635..3654
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3768..3872
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3874..4231
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3874..4065
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3897..3917)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4082..4217
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4266..5060
                     /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             4266..5060
                     /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(4320..4339)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4930..4949
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5248..5267)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5292..5339
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5668..6256
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6157..6176
                     /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 cgggatccac cggtcatggt gagcaagggc
     2341 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     2401 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     2461 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     2521 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     2581 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     2641 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     2701 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     2761 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     2821 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     2881 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     2941 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     3001 cactccaccg gcggcatgga cgagctgtac aagtaggcgg ccgcgactct agatcataat
     3061 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     3121 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3181 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3241 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     3301 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3361 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3421 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3481 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3541 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3601 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3661 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3721 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3781 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3841 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     3901 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3961 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4021 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4081 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4141 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4201 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     4261 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4321 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4381 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4441 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4501 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4561 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4621 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4681 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4741 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     4801 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4861 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4921 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4981 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5041 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     5101 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5161 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5221 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     5281 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     5341 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     5401 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     5461 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     5521 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5581 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5641 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5701 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5761 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5821 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5881 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5941 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6001 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6061 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6121 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6181 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6241 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6301 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6361 cgccatgcat
//
