LOCUS       sequence-140072-        6897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express N-terminal part (1-I210) of hTRAFD1 (FLN29) in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6897)
  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 6897)
  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-N/pCMV-14-FLAG (1-I210)
FEATURES             Location/Qualifiers
     source          1..6897
                     /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             1580..1645
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1656..2278
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1855..1874)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2302..2322)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2320..2617
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2671..3465
                     /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(2725..2744)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3335..3354
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4117..4251
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4154..4173)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4208..4227
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4256..4275)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4257..4275)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4289..4305)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4289..4305)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4302..4324)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4313..4329
                     /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(4337..4343,4344..4361,4362..4367))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4382..4403
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4520..4537)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4691..5279)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4771..4790)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5450..6241,6242..6310))
                     /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     6073..6092
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6311..6415)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6442..6870
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6529..6548)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6739..6760
                     /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 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 ttcaattgga tcccgggctg actacaaaga ccatgacggt gattataaag atcatgacat
     1621 cgactacaag gatgacgatg acaagtagtg atcccgggtg gcatccctgt gacccctccc
     1681 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
     1741 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
     1801 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
     1861 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
     1921 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca
     1981 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
     2041 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
     2101 aggcgtgaac cactgctccc ttccctgtcc ttctgatttt aaaataacta taccagcagg
     2161 aggacgtcca gacacagcat aggctacctg gccatgccca accggtggga catttgagtt
     2221 gcttgcttgg cactgtcctc tcatgcgttg ggtccactca gtagatgcct gttgaattgg
     2281 gtacgcggcc agcttggctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2341 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     2401 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2461 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2521 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     2581 ctgagctatt ccagaagtag tgaggaggct tttttggagg aattgatcag cttgggatct
     2641 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     2701 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     2761 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     2821 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     2881 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     2941 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3001 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3061 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3121 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3181 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3241 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3301 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3361 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3421 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     3481 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     3541 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     3601 agcgcgggga tctcatgctg gagttcttcg cccaccccgg gctcgatccc ctcgcgagtt
     3661 ggttcagctg ctgcctgagg ctggacgacc tcgcggagtt ctaccggcag tgcaaatccg
     3721 tcggcatcca ggaaaccagc agcggctatc cgcgcatcca tgcccccgaa ctgcaggagt
     3781 ggggaggcac gatggccgct ttggtcgacc cggacgggac gctcctgcgc ctgatacaga
     3841 acgaattgct tgcaggcatc tcatgagtgt gtcttcccgt tttccgcctg aggtcactgc
     3901 gtggatggag cgctggcgcc tgctgcgcga cggcgagctg ctcaccaccc actcgccaag
     3961 ctggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc gatcataatc
     4021 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     4081 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     4141 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4201 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat caattcccta
     4261 tagtgagtcg tattaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc
     4321 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4381 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4441 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4501 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4561 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4621 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4681 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4741 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4801 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4861 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4921 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4981 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5041 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5101 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5161 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5221 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5281 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5341 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5401 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5461 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5521 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5581 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5641 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5701 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5761 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5821 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5881 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5941 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6001 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6061 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6121 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6181 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6241 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6301 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6361 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6421 ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     6481 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     6541 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     6601 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     6661 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     6721 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     6781 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     6841 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaattt
//
