LOCUS       sequence-396912-        6344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mutated Translocon-associated protein 
            subunit alpha.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6344)
  AUTHORS   Karki S, Javanainen M, Rehan S, Tranter D, Kellosalo J, Huiskonen 
            JT, Happonen L, Paavilainen V
  TITLE     Molecular view of ER membrane remodeling by the Sec61/TRAP 
            translocon.
  JOURNAL   EMBO Rep. 2023 Dec 6;24(12):e57910. doi: 10.15252/embr.202357910. 
            Epub 2023 Nov 20.
  PUBMED    37983950
REFERENCE   2  (bases 1 to 6344)
  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: "EMBO Rep.";
            date: "2023-12-6"; volume: "24(12):e57910. doi"; pages: " 
            10.15252/embr.202357910. Epub 2023 Nov 20"
COMMENT     Sequence Label: TRAPa(R150A)-3xFLAG-pCDNA
FEATURES             Location/Qualifiers
     source          1..6344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..25
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(244..348)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(423..442)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        614..993
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        994..1197
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1148..1168
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1242..1261
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1242..1260
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2201..2266
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2314..2331)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2320..2544
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2590..3018
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2677..2696)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2887..2908
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3027..3047)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3032..3361
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3212..3347
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3274..3293
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3428..4222
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3482..3501)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4092..4111
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4396..4517
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4433..4452)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4487..4506
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4566..4582)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4566..4582)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4579..4601)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4590..4606
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4614..4620,4621..4638,4639..4644))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4659..4680
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4797..4814)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4968..5556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5048..5067)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5727..6344,1..174,175..243))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
       61 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
      121 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
      181 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
      241 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
      301 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
      361 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
      421 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
      481 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
      541 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
      601 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      661 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      721 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      781 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      841 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      901 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      961 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     1021 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     1081 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     1141 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
     1201 tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg
     1261 gagacccaag ctggctagcg tttaaactta agcttggtac cgagctcgga tccactagtc
     1321 cagtgtggtg gaattcgcca ccatgagact cctcccccgc ttgctgctgc ttctcttact
     1381 cgtgttccct gccactgtct tgttccgagg cggccccaga ggcttgttag cagtggcaca
     1441 agatcttaca gaggatgaag aaacagtaga agattccata attgaggatg aagatgatga
     1501 agccgaggta gaagaagatg aacccacaga tttggtagaa gataaagagg aagaagatgt
     1561 gtctggtgaa cctgaagctt caccgagtgc agatacaact atactgtttg taaaaggaga
     1621 agattttcca gcaaataaca ttgtgaagtt cctggtaggc tttaccaaca agggtacaga
     1681 agattttatt gttgaatcct tagatgcctc attccgttat cctcaggact accagtttta
     1741 tatccagaat ttcacagctc ttcctctgaa cactgtagtg ccaccccagg cacaggcaac
     1801 ttttgagtac tctttcattc ctgcagagcc catgggcgga cgaccatttg gtttggtcat
     1861 caatctgaac tacaaagatt tgaacggcaa tgtattccaa gatgcagtct tcaatcaaac
     1921 agttacagtt attgaaagag aggatgggtt agatggagaa acaatcttta tgtatatgtt
     1981 ccttgctggt cttgggcttc tggttattgt tggccttcat caactcctag aatctagaaa
     2041 gcgtaagaga cccatacaga aagtagaaat gggtacatca agtcagaatg atgttgacat
     2101 gagttggatt cctcaggaaa cattgaatca aatcaataaa gcttcaccaa gaaggttgcc
     2161 caggaaacgg gcacagaaga gatcagtggg atctgatgag gactataaag atcatgacgg
     2221 tgattacaag gatcacgata tcgactataa agatgacgac gacaagtagg cggccgctcg
     2281 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     2341 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     2401 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     2461 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     2521 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     2581 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2641 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     2701 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2761 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     2821 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2881 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     2941 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3001 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3061 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3121 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3181 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3241 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3301 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3361 aaagctcccg ggagcttata tatccatttt cggatctgat caagagacag gatgaggatc
     3421 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3481 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3541 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3601 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3661 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3721 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3781 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3841 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3901 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     3961 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4021 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4081 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4141 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4201 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4261 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4321 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4381 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4441 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4501 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     4561 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4621 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4681 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4741 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4801 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4861 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4921 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4981 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5041 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5101 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5161 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5221 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5281 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5341 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5401 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5461 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5521 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5581 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5641 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5701 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5761 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5821 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5881 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5941 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6001 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6061 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6121 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6181 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6241 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6301 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caat
//
