LOCUS       sequence-181639-        4945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains expanded CGG repeats (98 repeats with 2 AGG interruptions) 
            within the 5'UTR of FMR1. To generate capillary electrophoresis size
            standards..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4945)
  AUTHORS   Hayward BE, Usdin K
  TITLE     Improved Assays for AGG Interruptions in Fragile X Premutation 
            Carriers.
  JOURNAL   J Mol Diagn. 2017 Nov;19(6):828-835. doi: 
            10.1016/j.jmoldx.2017.06.008. Epub 2017 Aug 14.
  PUBMED    28818679
REFERENCE   2  (bases 1 to 4945)
  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: 
            "10.1016/j.jmoldx.2017.06.008"; journalName: "J Mol Diagn"; date: 
            "2017-11"; volume: "19"; issue: "6"; pages: "828-835"
COMMENT     Sequence Label: pGLACTE-HT51-10
FEATURES             Location/Qualifiers
     source          1..4945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(136..153)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(307..895)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(387..406)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1066..1857,1858..1926))
                     /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     1689..1708
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1927..2031)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2058..2513
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2145..2164)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2355..2376
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     protein_bind    2744..2812
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     protein_bind    3008..3026
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3044..3062
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3080..3098
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3116..3134
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3152..3170
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3188..3206
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3224..3242
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3287..3311
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(3890..3892,3893..3895,3896..4606)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3890..3892,3893..3895,3896..4606)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3935..3956)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4196..4215)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4543..4564
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4607..4672
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    4796..4917
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4833..4852)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4887..4906
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact
       61 atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca
      121 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
      181 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
      241 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      301 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      361 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      421 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      481 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      541 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      601 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      661 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      721 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
      781 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      841 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
      901 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
      961 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1021 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1081 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1141 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1201 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1261 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1321 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1381 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1441 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1501 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1561 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1621 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1681 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1741 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1801 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1861 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     1921 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     1981 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2041 ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2101 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2161 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2221 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2281 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2341 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     2401 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     2461 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttgccattc
     2521 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     2581 ccagcccaag ctaccatgat aagtaagtaa tattaaggta cgggaggtac ttggagcggc
     2641 cgcggtcagg accttttata tcggcctcta tagtctcatc tcttggtctc ggctgtgttc
     2701 ggtgggacgc ttgcgatttg ctgatatgta atttcaagtt attctcgaag ccgcggtgcg
     2761 ggtgccaggg cgtgcccttg ggctccccgg gcgcgtactc cacctcaccc atttcatttt
     2821 ggggttgcta atgcgcgtag gatacagaga gccagctctc acgatagccg taagtttttg
     2881 cgcctatgcc caagtatgac gcacttcgtt tccgtggcca ccagggggcg ccggtggccg
     2941 ccagggggca ccggtggcca ccagggggcg cctgaggctt tcgtcttcaa gaattcctcg
     3001 agtttactcc ctatcagtga tagagaacgt atgaagagtt tactccctat cagtgataga
     3061 gaacgtatgc agactttact ccctatcagt gatagagaac gtataaggag tttactccct
     3121 atcagtgata gagaacgtat gaccagttta ctccctatca gtgatagaga acgtatctac
     3181 agtttactcc ctatcagtga tagagaacgt atatccagtt tactccctat cagtgataga
     3241 gaacgtataa gctttaggcg tgtacggtgg gcgcctataa aagcagagct cgtttagtga
     3301 accgtcagat cgcctggagc aattccggcg cgccgctcag ctccgtttcg gtttcacttc
     3361 cggtggaggg ccgcctctga gcgggcggcg ggccgacggc gagcgcgggc ggcggcggtg
     3421 acggaggcgc cgctgccagg gggcgtgcgg cagcgcggcg gcggcggcgg cggcggcggc
     3481 ggaggcggcg gcggcggcgg cggcggcggc ggaggcggcg gcggcggcgg cggcggcggc
     3541 ggcggcggcg gcggcggcgg cggcggcggc ggcggcggcg gcggcggcgg cggcggcggc
     3601 ggcggcggcg gcggcggcgg cggcggcggc ggcggcggcg gcggcggcgg cggcggcggc
     3661 ggcggcggcg gcggcggcgg cggcggcggc ggcggcggcg gcggcggcgg cggcggcggc
     3721 ggcggcggcg gcggcggcgg cggcggcggc tgggcctcga gcgcccgcag cccacctctc
     3781 gggggcgggc tcccggcgct agcagggctg aagagaagat ggaggagctg gtggtggaag
     3841 tgcggggctc caatggagct tctacaaggc atttgtaaag gatgttccca tggtgagcaa
     3901 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     3961 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     4021 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     4081 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     4141 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     4201 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     4261 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     4321 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     4381 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     4441 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     4501 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     4561 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtccg gactcagatc
     4621 tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc
     4681 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     4741 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     4801 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4861 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg
     4921 cgtaaattgt aagcgttgcg gccgc
//
