LOCUS       sequence-464761-        6710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mini FXN plasmid with a partial deletion of the 5' UTR and intron 1 
            in FXN. Does not contain GAA repeats..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6710)
  AUTHORS   Li J, Li Y, Wang J, Gonzalez TJ, Asokan A, Napierala JS, Napierala M
  TITLE     Defining Transcription Regulatory Elements in the Human Frataxin 
            Gene: Implications for Gene Therapy.
  JOURNAL   Hum Gene Ther. 2020 Aug;31(15-16):839-851. doi: 
            10.1089/hum.2020.053. Epub 2020 Jul 13.
  PUBMED    32527155
REFERENCE   2  (bases 1 to 6710)
  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: "Hum Gene 
            Ther."; date: "2020-08"; volume: "31(15-16):839-851. doi"; pages: " 
            10.1089/hum.2020.053. Epub 2020 Jul 13"
COMMENT     Sequence Label: pCR4_miniFXN_7
FEATURES             Location/Qualifiers
     source          1..6710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    1044..1178
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1297..1320)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3574..3708
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3852..3871)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3853..3871)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3878..3895)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3878..3894)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3887..3909)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             4032..4334
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4213..4232
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             4683..5477
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4683..5477
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4737..4756)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5347..5366
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(5727..6518,6519..6587))
                     /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     6350..6369
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctca gaattaaccc
     1021 tcactaaagg gactagagtc gcagatccag acatgataag atacattgat gagtttggac
     1081 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     1141 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     1201 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     1261 aatgtggtat ggctgattat gatcctctag atatcactta tcgtcgtcat ccttgtaatc
     1321 catcaccagt atccgagcat cttttccgga ataggccaag gaagacaagt ccagtttggt
     1381 ttttaaggct ttagtgagct ctgcggccag cagctcatgg agggacacgc cgtcgtggga
     1441 gtacacccag tttttcccag tccagtcata acgcttaggt ccactggatg gagaagatag
     1501 ccagatttgc ttgtttggcg tctgcttgtt gatcacatag gttcctagat ctccacccag
     1561 tttgacagtt aagacaccac tcccaaagga gacatcatag tcctcaaacg tgtatggctt
     1621 gtctgcaagg tcttcaaaaa actctgctaa agagtccagc gtttcctctg ctagtctttc
     1681 ataggtggtc tcatctagag agcctgggtg gcccaaagtt ccagatttcc tcaaattcat
     1741 caaatagaca ctctgctttt tgacattcca aatctggttg aggccacgtt ggttcgaact
     1801 ctgaaagcca aagttaagaa gtacgttact atttttcagt gaattataac ccatgtgtgg
     1861 gaagtttaag cctaaagtac aaactccgga gagcaacaca aatatggctt ggacgtggcc
     1921 tgcctctttc atctccccta atacatgcgg cgtaccagcc actctgaagg gatccccttc
     1981 cgccttcctg gcctttgccc agacggttcc ctcctcgtga aacaccctct accacttcct
     2041 tggcatcttc aagaccctca ttaaatacca tgtcctcccc ttgaggaatc ttcctcatcc
     2101 ccacagccat tctttgggtt tcctcctttc aagccgtggc gtaactgggt cagatacttc
     2161 tttgtacccc ccaaaggaag aaaggggaac taatttgaag ttctgaagac tttatctatg
     2221 ttattcccat ttaatcctca caccaggtcc gcaaaatggg cgtcaccttt atcttctccc
     2281 tacagaaaac aaagccacgt ctcagagagg ttaggggaat cccccaaggt cacacagctc
     2341 tgcggagtgg ggcagaatct ggaataaagg tcgggtcagt ttccaaaagc cagggcccct
     2401 gctgctctag ctattctgca gctctgaaag tttcacctcg ttccaggaaa gcatttcgtt
     2461 caaaaaaatc cagccttgaa aatattaggt gtgcgcgcgc ccgtgcgcgc aaacacacac
     2521 acacaacaca cacgctatat acagatacac acagacacac acattatgtg tatgtatata
     2581 ctagcagata ccatctatta atattactga aacaagttat cgcagagaag tgacaagcat
     2641 ggagacagcc gcacacccct cggaaccggt cccctccacc agcctgccct gccttcgccc
     2701 gtgccttgac ccgcagtcgc accgcaggac aaaatgtccc cttttccttc ggaaagcaga
     2761 agccagtgta aatgcaaccg ggagaaccag agaagggagt tgcaaggccg cttccgccgg
     2821 gccgccctga gaaggagcgg ggtgagctag tccagcgcgc gtacccggag cgaccccggc
     2881 gtgcgcggcg cctccctgtg caggcgtgcg gcgtgcggcc cgcggcgtgc ggcccgcggc
     2941 tgttcccggc gcggatactt actgcgcggc ggggcgtgca ggtcgcatcg atgtcggtgc
     3001 gcaggccacg gcggccgcag agtggggcca actctgccgg ccgcgggacc cgggtgaggg
     3061 tctgggcctg ggccgggctg ggtgacgcca ggaggccggc tactgcgcgg cgcccgagag
     3121 tccacatgct gctccgggtc tgccgcccgc tccgccctcc agcgctgggt gctgcggcga
     3181 cccctggtgg ccactggccg caggcactct tctgtggggg agcagctaga ggttagacct
     3241 caggaagaac ttcccagctt agcactattc gtgcatttaa caaaaatgga gagcctgctt
     3301 tgtgcaaagc acggagtgca accaggaccc ctgacccaag ggagactgca gcctggtggc
     3361 ccgccgcttc caaaattcta aacaggagga acttgggagc tgctgtcttg ctgggaagtg
     3421 ggtgcatgca caaattgagg ctgcttggcc gccggtatgg gtttacagca gttgggtatg
     3481 tggggccagg agacggatgc cttgtgtaaa tctgtactct gcccacttcc tagctgtgtg
     3541 accagagggc aagttaaatt cactagagtc gcagatccag acatgataag atacattgat
     3601 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     3661 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     3721 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     3781 aacctctaca aatgtggtat ggctgattat gatcctctag ttacgacgtc gcggccgcta
     3841 aattcaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc
     3901 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     3961 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     4021 tatacgtacg gcagtttaag gtttacacct ataaaagaga gagccgttat cgtctgtttg
     4081 tggatgtaca gagtgatatt attgacacgc cggggcgacg gatggtgatc cccctggcca
     4141 gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg catatcgggg
     4201 atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc gttatcgggg
     4261 aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt aacctgatgt
     4321 tctggggaat ataaatgtca ggcatgagat tatcaaaaag gatcttcacc tagatccttt
     4381 tcacgtagaa agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg
     4441 ctatctggac aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta
     4501 catggcgata gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg
     4561 gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc
     4621 caaggatctg atggcgcagg ggatcaagct ctgatcaaga gacaggatga ggatcgtttc
     4681 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4741 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4801 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4861 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4921 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4981 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5041 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5101 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5161 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     5221 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5281 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5341 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5401 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5461 ttgacgagtt cttctgaatt attaacgctt acaatttcct gatgcggtat tttctcctta
     5521 cgcatctgtg cggtatttca caccgcatca ggtggcactt ttcggggaaa tgtgcgcgga
     5581 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagattatca
     5641 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5701 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5761 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5821 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5881 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5941 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6001 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6061 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6121 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6181 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6241 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6301 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6361 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6421 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6481 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6541 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6601 caatattatt gaagcattta tcagggttat tgtctcatga ccaaaatccc ttaacgtgag
     6661 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
