LOCUS       sequence-181640-        4742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains normal CGG repeats (29 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 4742)
  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 4742)
  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-HT51-13
FEATURES             Location/Qualifiers
     source          1..4742
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(140..157)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(311..899)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(391..410)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1070..1861,1862..1930))
                     /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     1693..1712
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1931..2035)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2062..2517
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2149..2168)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2359..2380
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     protein_bind    2748..2816
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     protein_bind    3012..3030
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3048..3066
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3084..3102
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3120..3138
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3156..3174
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3192..3210
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3228..3246
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3291..3315
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(3687..3689,3690..3692,3693..4403)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3687..3689,3690..3692,3693..4403)
                     /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(3732..3753)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3993..4012)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4340..4361
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4404..4469
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    4593..4714
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4630..4649)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4684..4703
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
       61 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
      121 ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
      181 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
      241 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
      301 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
      361 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
      421 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
      481 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
      541 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
      601 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
      661 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
      721 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
      781 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
      841 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
      901 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
      961 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     1021 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     1081 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     1141 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     1201 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     1261 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     1321 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     1381 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     1441 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     1501 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     1561 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     1621 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     1681 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     1741 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     1801 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     1861 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     1921 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     1981 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     2041 ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2101 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     2161 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2221 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     2281 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     2341 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     2401 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     2461 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttgcc
     2521 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     2581 tacgccagcc caagctacca tgataagtaa gtaatattaa ggtacgggag gtacttggag
     2641 cggccgcggt caggaccttt tatatcggcc tctatagtct catctcttgg tctcggctgt
     2701 gttcggtggg acgcttgcga tttgctgata tgtaatttca agttattctc gaagccgcgg
     2761 tgcgggtgcc agggcgtgcc cttgggctcc ccgggcgcgt actccacctc acccatttca
     2821 ttttggggtt gctaatgcgc gtaggataca gagagccagc tctcacgata gccgtaagtt
     2881 tttgcgccta tgcccaagta tgacgcactt cgtttccgtg gccaccaggg ggcgccggtg
     2941 gccgccaggg ggcaccggtg gccaccaggg ggcgcctgag gctttcgtct tcaagaattc
     3001 ctcgagttta ctccctatca gtgatagaga acgtatgaag agtttactcc ctatcagtga
     3061 tagagaacgt atgcagactt tactccctat cagtgataga gaacgtataa ggagtttact
     3121 ccctatcagt gatagagaac gtatgaccag tttactccct atcagtgata gagaacgtat
     3181 ctacagttta ctccctatca gtgatagaga acgtatatcc agtttactcc ctatcagtga
     3241 tagagaacgt ataagcttta ggcgtgtacg gtgggcgcct ataaaagcag agctcgttta
     3301 gtgaaccgtc agatcgcctg gagcaattcc ggcgcgccgc tcagctccgt ttcggtttca
     3361 cttccggtgg agggccgcct ctgagcgggc ggcgggccga cggcgagcgc gggcggcggc
     3421 ggtgacggag gcgccgctgc cagggggcgt gcggcagcgc ggcggcggcg gcggcggcgg
     3481 cggcggaggc ggcggcggcg gcggcggcgg cggcggaggc ggcggcggcg gcggcggcgg
     3541 cggcggctgg gcctcgagcg cccgcagccc acctctcggg ggcgggctcc cggcgctagc
     3601 agggctgaag agaagatgga ggagctggtg gtggaagtgc ggggctccaa tggcgctttt
     3661 acaaggcatt tgtaaaggat gttcccatgg tgagcaaggg cgaggagctg ttcaccgggg
     3721 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     3781 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     3841 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     3901 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     3961 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     4021 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     4081 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     4141 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     4201 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     4261 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     4321 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     4381 tcggcatgga cgagctgtac aagtccggac tcagatctcg agctcaagct tcgaattctg
     4441 cagtcgacgg taccgcgggc ccgggatcca ccggatctag ataactgatc ataatcagcc
     4501 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     4561 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     4621 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4681 gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttgcggcc
     4741 gc
//
