LOCUS       sequence-319466-        5906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  donor plasmid for C-terminal FLAG tag fusion of human FMR1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5906)
  AUTHORS   Li M, Shin J, Risgaard RD, Parries MJ, Wang J, Chasman D, Liu S, Roy
            S, Bhattacharyya A, Zhao X
  TITLE     Identification of FMR1-regulated molecular networks in human 
            neurodevelopment.
  JOURNAL   Genome Res. 2020 Mar;30(3):361-374. doi: 10.1101/gr.251405.119. Epub
            2020 Mar 16.
  PUBMED    32179589
REFERENCE   2  (bases 1 to 5906)
  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"; doi: 
            "10.1101/gr.251405.119"; journalName: "Genome Res"; date: "2020-03";
            volume: "30"; issue: "3"; pages: "361-374"
COMMENT     Sequence Label: FMR1-E17C-3xFLAG
FEATURES             Location/Qualifiers
     source          1..5906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     132..151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     385..402
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    519..540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(555..560,561..578,579..585)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    593..609
                     /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)."
     primer_bind     598..620
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     617..633
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     617..633
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1741..1806
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2751..2770)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2752..2770)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2777..2794)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2777..2793)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2786..2808)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2934..3362
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3021..3040)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3231..3252
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3706..4500
                     /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(3760..3779)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4370..4389
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4518..4586,4587..5378)
                     /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     complement(4736..4755)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5549..5906,1..231)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
       61 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      121 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      181 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      241 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      301 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      361 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      421 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      481 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
      541 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      601 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct tggtaccgag
      661 ctcggatcca catgcacata cccactactt aaacatactt tgaaattaca accatttggg
      721 gatgttttta gcatttgtgc ttgaagtaga tcaatatttg tagttgtttt agttccattt
      781 gtcactgtta actttcattt gtacctctgg aattagcagt gctgtattca gcattggcac
      841 ttaaaatatt ttatagctct tagaacacta ttttttaaat tacattgatc tttatactct
      901 cctaggattc cagtccttcc acaaattatt atatgcagtt ttctttatac tgttccattc
      961 ttcagacaga gttcaggttg taggatcatc tgaattttct tagtgaaata caagccagta
     1021 gaaaccaaag ctcgtgtttt tctactgtat gactatggga gctctcagta tagagaaact
     1081 tgatcatacc tgtgatagat ataaagatgt aaattaaagt ctagttagtg agttatttga
     1141 gggtaaagcc agaataggaa aacattttgt ttctcttttc ctgggcttca caatctttgg
     1201 agaataacta attttatttt aactctagtg ttagaggcat gtatgtcttc tggtaatcag
     1261 taatagtaat tcattttacc tgtttgtttc tcactgtttt attctactta gaagaagaca
     1321 tgataggatt gtgagttttt ctctaacttt ggatgcagtg agatgaccag tgtgttccag
     1381 ttaaagaaga agagtgtttt aaaatcataa accaaataaa gaatcctacc ttacattaat
     1441 ttcttgcact tcttctgttc tcatctccat ttctcttttt aacatgtgga atttacactt
     1501 caggtttaaa tttcttgtca ggccaattac agattacagt aggatatggt ctgtgtatat
     1561 aacaactata acttgtttta gatcagagtt gactgcaata atgaaaggag tgtccacact
     1621 aaaacattac agaatacctc cagtgaaggt agtcggctgc gcacgggtaa agatcgtaac
     1681 cagaagaaag agaagccaga cagcgtggat ggtcagcaac cactcgtgaa tggagtaccc
     1741 gattacaaag accatgacgg tgactataaa gatcatgaca tcgattacaa ggatgacgat
     1801 gacaagtaaa ctgcataatt ctgaagttat atttcctata ccatttccgt aattcttatt
     1861 ccatattaga aaactttgtt aggccaaaga caaatagtag gcaagatggc acagggcatg
     1921 aaatgaacac aaattatgct aagaattttt tattttttgg tattggccat aagcaacaat
     1981 tttcagattt gcacaaaaag ataccttaaa atttgaaaca ttgcttttaa aactacttag
     2041 cacttcaggg cagattttag ttttattttc taaagtactg agcagtgata ttctttgtta
     2101 atttggacca ttttcctgca ttgggtgatc attcaccagt acattctcag tttttcttaa
     2161 tatatagcat ttatggtaat catattagac ttctgttttc aatctcgtat agaagtcttc
     2221 atgaaatgct atgtcatttc atgtcctgtg tcagtttatg ttttggtcca cttttccagt
     2281 attttagtgg accctgaaat gtgtgtgatg tgacatttgt cattttcatt agcaaaaaaa
     2341 gttgtatgat ctgtgccttt tttatatctt ggcaggtagg aatattatat ttggatgcag
     2401 agttcaggga agataagttg gaaacactaa atgttaaaga tgtagcaaac cctgtcaaac
     2461 attagtactt tatagaagaa tgcatgcttt ccatattttt ttccttacat aaacatcagg
     2521 ttaggcagta taaagaatag gacttgtttt tgtttttgtt ttgttgcact gaagtttgat
     2581 aaatagtgtt attgagagag atgtgtaatt tttctgtata gacaggagaa gaaagaacta
     2641 tcttcatctg agagaggcta aaatgttttc agctaggaac aaatcttcct ggtcgaaagt
     2701 tagtaggata tgcctggccg ctcgagcatg catctagagg gcccaattcg ccctatagtg
     2761 agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     2821 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     2881 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc
     2941 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3001 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3061 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3121 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3181 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3241 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3301 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3361 ttttaacaaa attcagggcg caagggctgc taaaggaagc ggaacacgta gaaagccagt
     3421 ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa
     3481 aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac
     3541 tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag
     3601 gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc
     3661 aggggatcaa gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3721 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3781 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3841 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3901 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3961 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatcc
     4021 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4081 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4141 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4201 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4261 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4321 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4381 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4441 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4501 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4561 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4621 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4681 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4741 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4801 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4861 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4921 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4981 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5041 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5101 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5161 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5221 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5281 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5341 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5401 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5461 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5521 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5581 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5641 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5701 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5761 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5821 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5881 gcacacagcc cagcttggag cgaacg
//
