LOCUS       sequence-140079-        6919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express middle part (L175-D390) of human TRAFD1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6919)
  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 6919)
  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: hFLN29-Mid/pCMV-14-FLAG (L175-D390)
FEATURES             Location/Qualifiers
     source          1..6919
                     /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             1601..1666
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1678..2300
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1877..1896)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2324..2344)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2342..2639
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2693..3487
                     /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(2747..2766)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3357..3376
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4139..4273
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4176..4195)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4230..4249
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4278..4297)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4279..4297)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4311..4327)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4311..4327)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4324..4346)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4335..4351
                     /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(4359..4365,4366..4383,4384..4389))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4404..4425
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4542..4559)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4713..5301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4793..4812)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5472..6263,6264..6332))
                     /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     6095..6114
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6333..6437)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6464..6892
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6551..6570)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6761..6782
                     /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 tcgtttagtg aaccgtcaga attaagcttg cggccgcatg ctggacccac ccatgaggct
      961 gccgcgaagg cccctgagag cctttgaatc agatgttttc cacaatagaa ctaccaacca
     1021 aaggaacatt acagcccagg tttcaattca gaataatctg tttgaagaac aagagaggca
     1081 ggaaaggaat agaggccaac agccccccaa agagggtggt gaagagagtg caaacttgga
     1141 cttcatgttg gccctaagtc tgcaaaatga aggccaagcc tccagtgtgg cagagcagga
     1201 cttctggagg gccgtatgtg aggccgacca gtctcatggc ggtcccaggt ctctcagtga
     1261 cataaagggt gcagctgacg agatcatgtt gccttgtgaa ttttgtgagg agctctaccc
     1321 agaggaactg ctgattgacc atcagacaag ctgtaaccct tcacgtgcct taccttcact
     1381 caatactggc agctcttccc ccagaggggt ggaggaacct gatgtcatct tccagaactt
     1441 cttgcaacag gctgcaagta accagttaga ctctttgatg ggcctgagca attcacaccc
     1501 tgtggaggag agcatcatta tcccatgtga attctgtggg gtacagctgg aagaggaggt
     1561 gctgttccat caccaggacc agtgtgacgg atcccgggct gactacaaag accatgacgg
     1621 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagg tgatcccggg
     1681 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     1741 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     1801 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     1861 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     1921 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     1981 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     2041 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     2101 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     2161 ttaaaataac tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc
     2221 caaccggtgg gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact
     2281 cagtagatgc ctgttgaatt gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta
     2341 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2401 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2461 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     2521 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     2581 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     2641 ggaattgatc agcttgggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     2701 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     2761 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     2821 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     2881 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     2941 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3001 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     3061 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     3121 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     3181 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     3241 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     3301 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     3361 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     3421 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     3481 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     3541 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     3601 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     3661 gggctcgatc ccctcgcgag ttggttcagc tgctgcctga ggctggacga cctcgcggag
     3721 ttctaccggc agtgcaaatc cgtcggcatc caggaaacca gcagcggcta tccgcgcatc
     3781 catgcccccg aactgcagga gtggggaggc acgatggccg ctttggtcga cccggacggg
     3841 acgctcctgc gcctgataca gaacgaattg cttgcaggca tctcatgagt gtgtcttccc
     3901 gttttccgcc tgaggtcact gcgtggatgg agcgctggcg cctgctgcgc gacggcgagc
     3961 tgctcaccac ccactcgcca agctggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4021 ataggctccg ccgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     4081 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     4141 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4201 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4261 tcatgtctgg atcaattccc tatagtgagt cgtattaaat tcgtaatcat gtcatagctg
     4321 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4381 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4441 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4501 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4561 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4621 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4681 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4741 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4801 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4861 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4921 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4981 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5041 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5101 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5161 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5221 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5281 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5341 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5401 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5461 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5521 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5581 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5641 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5701 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5761 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5821 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5881 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5941 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6001 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6061 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6121 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6181 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6241 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6301 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6361 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6421 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
     6481 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     6541 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     6601 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     6661 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     6721 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     6781 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     6841 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     6901 atattaacgc ttacaattt
//
