LOCUS       sequence-253051-        7059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 3xFlag dsykerin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7059)
  AUTHORS   Egan ED, Collins K
  TITLE     Specificity and stoichiometry of subunit interactions in the human 
            telomerase holoenzyme assembled in vivo.
  JOURNAL   Mol Cell Biol. 2010 Jun;30(11):2775-86. doi: 10.1128/MCB.00151-10. 
            Epub 2010 Mar 29.
  PUBMED    20351177
REFERENCE   2  (bases 1 to 7059)
  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: 
            "10.1128/MCB.00151-10"; journalName: "Mol Cell Biol"; date: 
            "2010-06"; volume: "30"; issue: "11"; pages: "2775-86"
COMMENT     Sequence Label: pcDNA3.1 3xF-dyskerin
FEATURES             Location/Qualifiers
     source          1..7059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     635..654
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        635..653
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             676..741
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2422..2439)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2428..2652
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2698..3126
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2785..2804)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2995..3016
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3135..3155)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3140..3469
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3320..3455
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3382..3401
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3536..4330
                     /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(3590..3609)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4200..4219
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4504..4625
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4541..4560)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4595..4614
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4674..4690)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4674..4690)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4687..4709)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4698..4714
                     /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)."
     promoter        complement(join(4722..4728,4729..4746,4747..4752))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4767..4788
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4905..4922)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5076..5664)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5156..5175)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5835..6626,6627..6695))
                     /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     6458..6477
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6696..6800)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6875..6894)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa
      601 ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc
      661 aagctggcta gcatggacta caaagaccat gacggtgatt ataaagatca tgacatcgat
      721 tacaaggatg acgatgacaa gaagcttatg gccatggagg ccccggggat ctacatggcg
      781 gatgcggaag taattatttt gccaaagaaa cataagaaga aaaaggagcg gaagtcattg
      841 ccagaagaag atgtagccga aatacaacac gctgaagaat ttcttatcaa acctgaatcc
      901 aaagttgcta agttggacac gtctcagtgg ccccttttgc taaagaattt tgataagctg
      961 aatgtaagga caacacacta tacacctctt gcatgtggtt caaatcctct gaagagagag
     1021 attggggact atatcaggac aggtttcatt aatcttgaca agccctctaa cccctcttcc
     1081 catgaggtgg tagcctggat tcgacggata cttcgggtgg agaagacagg gcacagtggt
     1141 actctggatc ccaaggtgac tggttgttta atcgtgtgca tagaacgagc cactcgcttg
     1201 gtgaagtcac aacagagtgc aggcaaagag tatgtgggga ttgtccggct gcacaatgct
     1261 attgaagggg ggacccagct ttctagggcc ctagaaactc tgacaggtgc cttattccag
     1321 cgacccccac ttattgctgc agtaaagagg cagctccgag tgaggaccat ctacgagagc
     1381 aaaatgattg aatacgatcc tgaaagaaga ttaggaatct tttgggtgag ttgtgaggct
     1441 ggcacctaca ttcggacatt atgtgtgcac cttggtttgt tattgggagt tggtggtcag
     1501 atgcaggagc ttcggagggt tcgttctgga gtcatgagtg aaaaggacca catggtgaca
     1561 atgcatgatg tgcttgatgc tcagtggctg tatgataacc acaaggatga gagttacctg
     1621 cggcgagttg tttacccttt ggaaaagctg ttgacatctc ataaacggct ggttatgaaa
     1681 gacagtgcag taaatgccat ctgctatggg gccaagatta tgcttccagg tgttcttcga
     1741 tatgaggacg gcattgaggt caatcaggag attgtggtta tcaccaccaa aggagaagca
     1801 atctgcatgg ctattgcatt aatgaccaca gcggtcatct ctacctgcga ccatggtata
     1861 gtagccaaga tcaagagagt gatcatggag agagacactt accctcggaa gtggggttta
     1921 ggtccaaagg caagtcagaa gaagctgatg atcaagcagg gccttctgga caagcatggg
     1981 aagcccactg acagcacacc tgccacctgg aagcaggagt atgttgacta cagtgagtct
     2041 gccaaaaaag aggtggttgc tgaagtggta aaagccccgc aggtagttgc cgaagcagca
     2101 aaaactgcga agcggaagcg agagagtgag agtgaaagtg acgagactcc tccagcagct
     2161 cctcagttga tcaagaagga aaagaagaag agtaagaagg acaagaaggc caaagctggt
     2221 ctggagagcg gggccgagcc tggagatggg gacagtgata ccaccaagaa gaagaagaag
     2281 aagaagaaag caaaagaggt agaattggtt tctgagtagt gatgaagatc cgaattcgag
     2341 ctcgagagat ccgaattcga gctccgtcga caagcttgcg gccgctcgag tctagagggc
     2401 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     2461 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2521 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2581 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     2641 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     2701 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2761 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2821 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2881 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     2941 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3001 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3061 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3121 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3181 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3241 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3301 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3361 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     3421 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     3481 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     3541 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3601 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3661 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     3721 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3781 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3841 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3901 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3961 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4021 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     4081 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4141 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4201 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4261 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4321 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4381 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4441 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4501 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4561 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4621 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     4681 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4741 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4801 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4861 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4921 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4981 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5041 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5101 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5161 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5221 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5281 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5341 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5401 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5461 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5521 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5581 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5641 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5701 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5761 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5821 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5881 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5941 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6001 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6061 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6121 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6181 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6241 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6301 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6361 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6421 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6481 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6541 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6601 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6661 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6721 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6781 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     6841 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     6901 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     6961 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     7021 ggcgttttgc gctgcttcgc gatgtacggg ccagatata
//
