LOCUS       sequence-445048-        5336 bp    DNA     circular SYN 15-APR-2025
DEFINITION  To create 0CGG FMR1 cell line.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5336)
  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 5336)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-dCGG0
FEATURES             Location/Qualifiers
     source          1..5336
                     /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(1769..1788)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1770..1788)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1795..1812)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1795..1811)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1804..1826)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1952..2380
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2039..2058)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2249..2270
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             2724..3518
                     /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(2778..2797)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3388..3407
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3536..3604,3605..4396)
                     /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(3754..3773)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4567..5155
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5056..5075
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5309..5326
                     /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 agcgctgggc ctcgagcgcc cgcagcccac ctctcggggg
     1081 cgggctcccg gcgctagcag ggctgaagag aagatggagg agctggtggt ggaagtgcgg
     1141 ggctccaatg gcgctttcta caaggtactt ggctctaggg caggccccat cttcgccctt
     1201 ccttccctcc cttttcttct tggtgtcggc gggaggcagg cccggggccc tcttcccgag
     1261 caccgcgcct gggtgccagg gcacgctcgg cgggatgttg ttgggaggga aggactggac
     1321 ttggggcctg ttggaagccc ctctccgact ccgagaggcc ctagcgccta tcgaaatgag
     1381 agaccagcga ggagagggtt ctctttcggc gccgagcccc gccggggtga gctggggatg
     1441 ggcgagggcc ggcggcaggt actagagccg ggcgggaagg gccgaaatcg gcgctaagtg
     1501 acggcgatgg cttattcccc ctttcctaaa catcatctcc cagcgggatc cgggcctgtc
     1561 gtgtgggtag ttgtggagga gcggggggcg cttcagccgg gccgcctcct gcagcgccaa
     1621 gagggcttca ggtctccttt ggcttctctt ttccggtcta gcattgggac ttcggagagc
     1681 tccactgttc tgggcgaggg ctgtgaagaa agagtagtaa gaagcggtag tcggcacgct
     1741 cgagcatgca tctagagggc ccaattcgcc ctatagtgag tcgtattaca attcactggc
     1801 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     1861 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     1921 ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg
     1981 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2041 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2101 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2161 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     2221 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2281 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     2341 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat tcagggcgca
     2401 agggctgcta aaggaagcgg aacacgtaga aagccagtcc gcagaaacgg tgctgacccc
     2461 ggatgaatgt cagctactgg gctatctgga caagggaaaa cgcaagcgca aagagaaagc
     2521 aggtagcttg cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa
     2581 gcgaaccgga attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa
     2641 actggatggc tttcttgccg ccaaggatct gatggcgcag gggatcaaga tctgatcaag
     2701 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     2761 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     2821 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     2881 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     2941 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3001 tattgggcga agtgccgggg caggatctcc tgtcatccca ccttgctcct gccgagaaag
     3061 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3121 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3181 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     3241 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3301 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     3361 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     3421 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     3481 gcatcgcctt ctatcgcctt cttgacgagt tcttctgaat tgaaaaagga agagtatgag
     3541 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3601 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3661 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3721 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3781 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3841 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3901 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3961 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4021 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4081 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4141 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4201 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4261 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4321 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4381 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4441 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4501 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4561 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4621 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4681 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4741 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4801 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4861 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4921 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4981 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5041 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5101 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5161 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5221 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     5281 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
