LOCUS       sequence-445290-        5356 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  0CGG with BsmBI site for inserting CGG repeats.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5356)
  AUTHORS   Sirois CL, Guo Y, Li M, Wolkoff NE, Korabelnikov T, Sandoval S, Lee 
            J, Shen M, Contractor A, Sousa AMM, Bhattacharyya A, Zhao X
  TITLE     CGG repeats in the human FMR1 gene regulate mRNA localization and 
            cellular stress in developing neurons.
  JOURNAL   Cell Rep. 2024 Jun 11;43(6):114330. doi: 
            10.1016/j.celrep.2024.114330.
  PUBMED    38865241
REFERENCE   2  (bases 1 to 5356)
  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: "Cell Rep.";
            date: "2024-06-11"; volume: "43(6):114330. doi"; pages: " 
            10.1016/j.celrep.2024.114330"
COMMENT     Sequence Label: pDonor-FMR1-CGG-BsmBI
FEATURES             Location/Qualifiers
     source          1..5356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1789..1808)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1790..1808)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1815..1832)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1815..1831)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1824..1846)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1972..2400
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2059..2078)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2269..2290
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             2744..3538
                     /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(2798..2817)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3408..3427
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3556..3624,3625..4416)
                     /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(3774..3793)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4587..5175
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5076..5095
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5329..5346
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 gtaccgagct cggagggtct gaggaaaggc tcacattttg agatcccgac tcaatccatg
      301 tcccttaaag ggcacagggt gtctccacag ggccgcccaa aatctggtga gagagggcgt
      361 agacgcctca ccttctgcct ctacgggtca caaaagcctg ggtcaccctg gttgccactg
      421 ttcctagttc aaagtcttct tctgtctaat ccttcacccc tattctcgcc ttccactcca
      481 cctcccgctc agtcagactg cgctactttg aaccggacca aaccaaacca aaccaaacca
      541 aaccaaacca gaccagacac cccctcccgc ggaatcccag agaggccgaa ctgggataac
      601 cggatgcatt tgatttccca cgccactgag tgcacctctg cagaaatggg cgttctggcc
      661 ctcgcgaggc agtgcgacct gtcaccgccc ttcagccttc ccgccctcca ccaagcccgc
      721 gcacgcccgg cccgcgcgtc tgtctttcga cccggcaccc cggccggttc ccagcagcgc
      781 gcatgcgcgc gctcccaggc cacttgaaga gagagggcgg ggccgagggg ctgagcccgc
      841 ggggggaggg aacagcgttg atcacgtgac gtggtttcag tgtttacacc cgcagcgggc
      901 cgggggttcg gcctcagtca ggcgctcagc tccgtttcgg tttcacttcc ggtggagggc
      961 cgcctctgag cgggcggcgg gccgacggcg agcgcgggcg gcggcggtga cggaggcgcc
     1021 gctgccaggg ggcgtgcggc agcgggagac gggataccgt ctcactgggc ctcgagcgcc
     1081 cgcagcccac ctctcggggg cgggctcccg gcgctagcag ggctgaagag aagatggagg
     1141 agctggtggt ggaagtgcgg ggctccaatg gcgctttcta caaggtactt ggctctaggg
     1201 caggccccat cttcgccctt ccttccctcc cttttcttct tggtgtcggc gggaggcagg
     1261 cccggggccc tcttcccgag caccgcgcct gggtgccagg gcacgctcgg cgggatgttg
     1321 ttgggaggga aggactggac ttggggcctg ttggaagccc ctctccgact ccgagaggcc
     1381 ctagcgccta tcgaaatgag agaccagcga ggagagggtt ctctttcggc gccgagcccc
     1441 gccggggtga gctggggatg ggcgagggcc ggcggcaggt actagagccg ggcgggaagg
     1501 gccgaaatcg gcgctaagtg acggcgatgg cttattcccc ctttcctaaa catcatctcc
     1561 cagcgggatc cgggcctgtc gtgtgggtag ttgtggagga gcggggggcg cttcagccgg
     1621 gccgcctcct gcagcgccaa gagggcttca ggtctccttt ggcttctctt ttccggtcta
     1681 gcattgggac ttcggagagc tccactgttc tgggcgaggg ctgtgaagaa agagtagtaa
     1741 gaagcggtag tcggcacgct cgagcatgca tctagagggc ccaattcgcc ctatagtgag
     1801 tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     1861 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     1921 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
     1981 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2041 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2101 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2161 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2221 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2281 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2341 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2401 ttaacaaaat tcagggcgca agggctgcta aaggaagcgg aacacgtaga aagccagtcc
     2461 gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga caagggaaaa
     2521 cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat agctagactg
     2581 ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct ctggtaaggt
     2641 tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct gatggcgcag
     2701 gggatcaaga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     2761 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     2821 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     2881 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     2941 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     3001 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatccca
     3061 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     3121 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     3181 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     3241 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     3301 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     3361 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     3421 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     3481 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgaat
     3541 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3601 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3661 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3721 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3781 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     3841 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3901 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3961 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4021 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     4081 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     4141 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4201 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     4261 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4321 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4381 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4441 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4501 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4561 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4621 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4681 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4741 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4801 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4861 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4921 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4981 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5041 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5101 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5161 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5221 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5281 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     5341 agcgaggaag cggaag
//
