LOCUS       sequence-140065-        8016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human TRAFD1 (FLN29) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8016)
  AUTHORS   Witwicka H, Jia H, Kutikov A, Reyes-Gutierrez P, Li X, Odgren PR
  TITLE     TRAFD1 (FLN29) Interacts with Plekhm1 and Regulates Osteoclast 
            Acidification and Resorption.
  JOURNAL   PLoS One. 2015 May 19;10(5):e0127537. doi: 
            10.1371/journal.pone.0127537. eCollection 2015.
  PUBMED    25992615
REFERENCE   2  (bases 1 to 8016)
  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: "PLoS One.";
            date: "2015-05-19"; volume: "10(5):e0127537. doi"; pages: " 
            10.1371/journal.pone.0127537. eCollection 2015"
COMMENT     Sequence Label: hTRAFD1-pCMV14-3xFlag
FEATURES             Location/Qualifiers
     source          1..8016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2699..2764
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2775..3397
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2974..2993)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3421..3441)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3439..3736
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3790..4584
                     /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(3844..3863)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4454..4473
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5236..5370
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5273..5292)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5327..5346
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5375..5394)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5376..5394)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5408..5424)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5408..5424)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5421..5443)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5432..5448
                     /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(5456..5462,5463..5480,5481..5486))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5501..5522
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5639..5656)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5810..6398)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5890..5909)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6569..7360,7361..7429))
                     /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     7192..7211
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7430..7534)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7561..7989
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7648..7667)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7858..7879
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttgtga accgtcagaa ttaagcttgc ggccgcgatg gctgaatttc tagatgacca
      961 ggaaactcga ctgtgtgaca actgcaaaaa agaaattcct gtgtttaact ttaccatcca
     1021 tgagatccac tgtcaaagga acattggtat gtgtcctacc tgtaaggaac catttcccaa
     1081 atctgacatg gagactcaca tggctgcaga acactgtcag gtgacctgca aatgtaacaa
     1141 gaagttggag aagaggctgt taaagaagca tgaggagact gagtgccctt tgcggcttgc
     1201 tgtctgccag cactgtgatt tagaactttc cattctcaaa ctgaaggaac atgaagatta
     1261 ttgtggtgcc cggacggaac tatgtggcaa ctgtggtcgc aatgtccttg tgaaagatct
     1321 gaagactcac cctgaagttt gtgggagaga gggggaggaa aagagaaatg aggttgccat
     1381 acctcctaat gcatatgatg aatcttgggg tcaggatgga atctggattg catcccaact
     1441 cctcagacaa attgaggctc tggacccacc catgaggctg ccgcgaaggc ccctgagagc
     1501 ctttgaatca gatgttttcc acaatagaac taccaaccaa aggaacatta cagcccaggt
     1561 ttcaattcag aataatctgt ttgaagaaca agagaggcag gaaaggaata gaggccaaca
     1621 gccccccaaa gagggtggtg aagagagtgc aaacttggac ttcatgttgg ccctaagtct
     1681 gcaaaatgaa ggccaagcct ccagtgtggc agagcaggac ttctggaggg ccgtatgtga
     1741 ggccgaccag tctcatggcg gtcccaggtc tctcagtgac ataaagggtg cagctgacga
     1801 gatcatgttg ccttgtgaat tttgtgagga gctctaccca gaggaactgc tgattgacca
     1861 tcagacaagc tgtaaccctt cacgtgcctt accttcactc aatactggca gctcttcccc
     1921 cagaggggtg gaggaacctg atgtcatctt ccagaacttc ttgcaacagg ctgcaagtaa
     1981 ccagttagac tctttgatgg gcctgagcaa ttcacaccct gtggaggaga gcatcattat
     2041 cccatgtgaa ttctgtgggg tacagctgga agaggaggtg ctgttccatc accaggacca
     2101 gtgtgaccaa cgcccagcca ctgcaaccaa ccatgtgaca gaggggattc ctagactgga
     2161 ttcccagcct caagagacct caccagagct gcccaggagg cgtgtcagac accagggaga
     2221 cctgtcttct ggttacctgg atgatactaa gcaggaaaca gctaatgggc ccacctcctg
     2281 tctgcctccc agccgaccca ttaacaatat gacagctacc tataaccagc tatcgagatc
     2341 aacatcaggc cccagacctg ggtgccagcc cagctctcct tgtgtgccga agctcagcaa
     2401 ctcagacagc caggacatcc aggggcggaa tcgagacagc cagaatgggg ccatagcccc
     2461 tgggcacgtt tcagtgattc gccctcctca aaatctctac ccagaaaaca ttgtgccctc
     2521 tttctcccct gggccttcag ggagatacgg agctagtggt aggagtgaag gtggcaggaa
     2581 ttcccgggtc acccctgcag ctgccaacta ccgcagcaga actgcaaagg caaagccttc
     2641 caagcaacag ggagctgggg atgcagaaga ggaagaggag gagcaaggat cccgggctga
     2701 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga
     2761 caagtagtga tcccgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg
     2821 aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt
     2881 ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc
     2941 aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag
     3001 tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc
     3061 agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt
     3121 tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg
     3181 tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct
     3241 tccctgtcct tctgatttta aaataactat accagcagga ggacgtccag acacagcata
     3301 ggctacctgg ccatgcccaa ccggtgggac atttgagttg cttgcttggc actgtcctct
     3361 catgcgttgg gtccactcag tagatgcctg ttgaattggg tacgcggcca gcttggctgt
     3421 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3481 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3541 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3601 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3661 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3721 gaggaggctt ttttggagga attgatcagc ttgggatctg atcaagagac aggatgagga
     3781 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3841 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3901 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3961 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4021 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4081 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4141 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4201 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4261 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4321 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4381 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4441 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4501 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4561 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4621 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4681 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4741 agttcttcgc ccaccccggg ctcgatcccc tcgcgagttg gttcagctgc tgcctgaggc
     4801 tggacgacct cgcggagttc taccggcagt gcaaatccgt cggcatccag gaaaccagca
     4861 gcggctatcc gcgcatccat gcccccgaac tgcaggagtg gggaggcacg atggccgctt
     4921 tggtcgaccc ggacgggacg ctcctgcgcc tgatacagaa cgaattgctt gcaggcatct
     4981 catgagtgtg tcttcccgtt ttccgcctga ggtcactgcg tggatggagc gctggcgcct
     5041 gctgcgcgac ggcgagctgc tcaccaccca ctcgccaagc tggaaccgta aaaaggccgc
     5101 gttgctggcg tttttccata ggctccgccg atcataatca gccataccac atttgtagag
     5161 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     5221 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5281 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5341 ctcatcaatg tatcttatca tgtctggatc aattccctat agtgagtcgt attaaattcg
     5401 taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5461 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5521 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5581 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5641 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5701 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5761 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5821 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5881 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5941 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6001 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6061 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6121 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6181 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6241 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6301 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6361 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6421 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6481 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6541 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6601 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6661 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6721 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6781 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6841 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6901 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6961 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7021 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7081 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7141 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7201 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7261 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7321 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7381 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7441 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7501 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7561 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     7621 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     7681 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     7741 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     7801 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     7861 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     7921 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     7981 acgcgaattt taacaaaata ttaacgctta caattt
//
