LOCUS       sequence-396915-        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(RKRKR232-236AAAAA)-3xFLAG-pCDNA
FEATURES             Location/Qualifiers
     source          1..6344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(116..220)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(295..314)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        486..865
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        866..1069
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1020..1040
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1114..1133
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1114..1132
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2073..2138
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2186..2203)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2192..2416
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2462..2890
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2549..2568)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2759..2780
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2899..2919)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2904..3233
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3084..3219
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3146..3165
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3300..4094
                     /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(3354..3373)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3964..3983
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4268..4389
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4305..4324)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4359..4378
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4438..4454)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4438..4454)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4451..4473)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4462..4478
                     /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(4486..4492,4493..4510,4511..4516))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4531..4552
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4669..4686)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4840..5428)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4920..4939)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5599..6344,1..46,47..115))
                     /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"
     primer_bind     6222..6241
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
       61 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
      121 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
      181 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
      241 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
      301 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
      361 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
      421 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
      481 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      541 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      601 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      661 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      721 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      781 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      841 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      901 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      961 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     1021 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
     1081 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
     1141 agctggctag cgtttaaact taagcttggt accgagctcg gatccactag tccagtgtgg
     1201 tggaattcgc caccatgaga ctcctccccc gcttgctgct gcttctctta ctcgtgttcc
     1261 ctgccactgt cttgttccga ggcggcccca gaggcttgtt agcagtggca caagatctta
     1321 cagaggatga agaaacagta gaagattcca taattgagga tgaagatgat gaagccgagg
     1381 tagaagaaga tgaacccaca gatttggtag aagataaaga ggaagaagat gtgtctggtg
     1441 aacctgaagc ttcaccgagt gcagatacaa ctatactgtt tgtaaaagga gaagattttc
     1501 cagcaaataa cattgtgaag ttcctggtag gctttaccaa caagggtaca gaagatttta
     1561 ttgttgaatc cttagatgcc tcattccgtt atcctcagga ctaccagttt tatatccaga
     1621 atttcacagc tcttcctctg aacactgtag tgccacccca gagacaggca acttttgagt
     1681 actctttcat tcctgcagag cccatgggcg gacgaccatt tggtttggtc atcaatctga
     1741 actacaaaga tttgaacggc aatgtattcc aagatgcagt cttcaatcaa acagttacag
     1801 ttattgaaag agaggatggg ttagatggag aaacaatctt tatgtatatg ttccttgctg
     1861 gtcttgggct tctggttatt gttggccttc atcaactcct agaatctgcg gcagctgccg
     1921 cgcccataca gaaagtagaa atgggtacat caagtcagaa tgatgttgac atgagttgga
     1981 ttcctcagga aacattgaat caaatcaata aagcttcacc aagaaggttg cccaggaaac
     2041 gggcacagaa gagatcagtg ggatctgatg aggactataa agatcatgac ggtgattaca
     2101 aggatcacga tatcgactat aaagatgacg acgacaagta ggcggccgct cgagtctaga
     2161 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2221 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2281 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2341 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2401 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     2461 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2521 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2581 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2641 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2701 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2761 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2821 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2881 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     2941 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3001 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3061 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3121 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3181 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3241 cgggagctta tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     3301 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3361 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3421 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3481 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3541 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3601 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3661 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3721 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3781 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     3841 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     3901 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     3961 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4021 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4081 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4141 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4201 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4261 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4321 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4381 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4441 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4501 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4561 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4621 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4681 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4741 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4801 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4861 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4921 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4981 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5041 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5101 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5161 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5221 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5281 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5341 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5401 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5461 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5521 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5581 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5641 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5701 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5761 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5821 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5881 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5941 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6001 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6061 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6121 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6181 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6241 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6301 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcac
//
