LOCUS       sequence-489462-        9069 bp    DNA     circular SYN 15-OCT-2025
DEFINITION  Retroviral expression of human dyskerin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9069)
  AUTHORS   Venteicher AS, Abreu EB, Meng Z, McCann KE, Terns RM, Veenstra TD, 
            Terns MP, Artandi SE
  TITLE     A human telomerase holoenzyme protein required for Cajal body 
            localization and telomere synthesis.
  JOURNAL   Science. 2009 Jan 30;323(5914):644-8. doi: 10.1126/science.1165357.
  PUBMED    19179534
REFERENCE   2  (bases 1 to 9069)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2009-01-30- 30"; volume: "323"; 
            issue: "5914"; pages: "644-8"
COMMENT     Sequence Label: pMGIB-3xFLAG-dyskerin
FEATURES             Location/Qualifiers
     source          1..9069
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     LTR             1517..1989
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    2054..2411
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             2484..2900
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2830..2852
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2868..2884
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     3214..3230
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     promoter        3260..3278
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      3339..3348
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3360..3425
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    5078..5539
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5134..5151)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5361..5380
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             6279..6748
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(7444..7461)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7444..7460)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7453..7475)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7669..7688)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7788..7810
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7848..7866)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7934..8038
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8039..8107,8108..8899)
                     /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(8257..8276)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttt gctcttagga
     1021 gtttcctaat acatcccaaa ctcaaatata taaagcattt gacttgttct atgccctagg
     1081 gggcgggggg aagctaagcc agcttttttt aacatttaaa atgttaattc cattttaaat
     1141 gcacagatgt ttttatttca taagggtttc aatgtgcatg aatgctgcaa tattcctgtt
     1201 accaaagcta gtataaataa aaatagataa acgtggaaat tacttagagt ttctgtcatt
     1261 aacgtttcct tcctcagttg acaacataaa tgcgctgctg agaagccagt ttgcatctgt
     1321 caggatcaat ttcccattat gccagtcata ttaattacta gtcaattagt tgatttttat
     1381 ttttgacata tacatgtgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg
     1441 ccattttgca aggcatggaa aaatacataa ctgagaatag aaaagttcag atcaaggtca
     1501 ggaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     1561 cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
     1621 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc
     1681 cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
     1741 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
     1801 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgccgcgc cagtcctccg
     1861 attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac
     1921 ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg
     1981 ggtctttcat ttgggggctc gtccgggatc gggagacccc tgcccaggga ccaccgaccc
     2041 accaccggga ggtaagctgg ccagcaactt atctgtgtct gtccgattgt ctagtgtcta
     2101 tgactgattt tatgcgcctg cgtcggtact agttagctaa ctagctctgt atctggcgga
     2161 cccgtggtgg aactgacgag ttcggaacac ccggccgcaa ccctgggaga cgtcccaggg
     2221 acttcggggg ccgtttttgt ggcccgacct gagtccaaaa atcccgatcg ttttggactc
     2281 tttggtgcac cccccttaga ggagggatat gtggttctgg taggagacga gaacctaaaa
     2341 cagttcccgc ctccgtctga atttttgctt tcggtttggg accgaagccg cgccgcgcgt
     2401 cttgtctgct gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct
     2461 gaaaatatgg gcccgccgcg gcgggccaga ctgttaccac tcccttaagt ttgaccttag
     2521 gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac
     2581 gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg
     2641 gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc
     2701 atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc
     2761 ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc
     2821 cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc
     2881 tcactccttc tctaggcgcc cccatatgag atcttatatg gggcaccccc gccccttgta
     2941 aacttccctg accctgacat gacaagagtt actaacagcc cctctctcca agctcactta
     3001 caggctctct acttagtcca gcacgaagtc tggagacctc tggcggcagc ctaccaagaa
     3061 caactggacc gaccggtggt acctcaccct taccgagtcg gcgacacagt gtgggtccgc
     3121 cgacaccaga ctaagaacct agaacctcgc tggaaaggac cttacacagt cctgctgacc
     3181 acccccaccg ccctcaaagt agacggcatc gcagcttgga tacacgccgc ccacgtaaaa
     3241 caattgaaga tctagtcgaa attaaccctc actaaaggga acaaaagctg gagctccacc
     3301 gcggtggcgg ccaattcgag ctcggtaccc gtttaaacgc caccatgggg tcccgggctg
     3361 actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg
     3421 acaaggggcg cgccgcggat gcggaagtaa ttattttgcc aaagaaacat aagaagaaaa
     3481 aggagcggaa gtcattgcca gaagaagatg tagccgaaat acaacacgct gaagaatttc
     3541 ttatcaaacc tgaatccaaa gttgctaagt tggacacgtc tcagtggccc cttttgctaa
     3601 agaattttga taagctgaat gtaaggacaa cacactatac acctcttgca tgtggttcaa
     3661 atcctctgaa gagagagatt ggggactata tcaggacagg tttcattaat cttgacaagc
     3721 cctctaaccc ctcttcccat gaggtggtag cctggattcg acggatactt cgggtggaga
     3781 agacagggca cagtggtact ctggatccca aggtgactgg ttgtttaatc gtgtgcatag
     3841 aacgagccac tcgcttggtg aagtcacaac agagtgcagg caaagagtat gtggggattg
     3901 tccggctgca caatgctatt gaagggggga cccagctttc tagggcccta gaaactctga
     3961 caggtgcctt attccagcga cccccactta ttgctgcagt aaagaggcag ctccgagtga
     4021 ggaccatcta cgagagcaaa atgattgaat acgatcctga aagaagatta ggaatctttt
     4081 gggtgagttg tgaggctggc acctacattc ggacattatg tgtgcacctt ggtttgttat
     4141 tgggagttgg tggtcagatg caggagcttc ggagggttcg ttctggagtc atgagtgaaa
     4201 aggaccacat ggtgacaatg catgatgtgc ttgatgctca gtggctgtat gataaccaca
     4261 aggatgagag ttacctgcgg cgagttgttt accctttgga aaagctgttg acatctcata
     4321 aacggctggt tatgaaagac agtgcagtaa atgccatctg ctatggggcc aagattatgc
     4381 ttccaggtgt tcttcgatat gaggacggca ttgaggtcaa tcaggagatt gtggttatca
     4441 ccaccaaagg agaagcaatc tgcatggcta ttgcattaat gaccacagcg gtcatctcta
     4501 cctgcgacca tggtatagta gccaagatca agagagtgat catggagaga gacacttacc
     4561 ctcggaagtg gggtttaggt ccaaaggcaa gtcagaagaa gctgatgatc aagcagggcc
     4621 ttctggacaa gcatgggaag cccacagaca gcacacctgc cacctggaag caggagtatg
     4681 ttgactacag tgagtctgcc aaaaaagagg tggttgctga agtggtaaaa gccccgcagg
     4741 tagttgccga agcagcaaaa actgcgaagc ggaagcgaga gagtgagagt gaaagtgacg
     4801 agactcctcc agcagctcct cagttgatca agaaggaaaa gaagaagagt aagaaggaca
     4861 agaaggccaa agctggtctg gagagcgggg ccgagcctgg agatggggac agtgatacca
     4921 ccaagaagaa gaagaagaag aagaaagcaa aagaggtaga attggtttct gagtagttaa
     4981 ttaagtctag agtcgactgt ttaaacctgc aggcatgcaa gctcgacctc gaccaattcc
     5041 ggttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     5101 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
     5161 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag
     5221 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     5281 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     5341 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     5401 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat
     5461 gtgtttagtc gaggttaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     5521 ttgaaaaaca cgatgataat accatgaaaa catttaacat ttctcaacag gatctagaat
     5581 tagtagaagt agcgacagag aagattacaa tgctttatga ggataataaa catcatgtgg
     5641 gagcggcaat tcgtacgaaa acaggagaaa tcatttcggc agtacatatt gaagcgtata
     5701 taggacgagt aactgtttgt gcagaagcca ttgcgattgg tagtgcagtt tcgaatggac
     5761 aaaaggattt tgacacgatt gtagctgtta gacaccctta ttctgacgaa gtagatagaa
     5821 gtattcgagt ggtaagtcct tgtggtatgt gtagggagtt gatttcagac tatgcaccag
     5881 attgttttgt gttaatagaa atgaatggca agttagtcaa aactacgatt gaagaactca
     5941 ttccactcaa atatacccga aattaaaagt tttaccatac caagctcatc gggccgccag
     6001 gtacaagtaa ggatccggat tagtccaatt tgttaaagac aggatatcag tggtccaggc
     6061 tctagttttg actcaacaat atcaccagct gaagcctata gagtacgagc catagataaa
     6121 ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt
     6181 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat
     6241 agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat
     6301 atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata
     6361 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     6421 tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg
     6481 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc
     6541 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac
     6601 tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc
     6661 tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga
     6721 ttgactaccc gtcagcgggg gtctttcaca tgcagcatgt atcaaaatta atttggtttt
     6781 ttttcttaag tatttacatt aaatggccat agtacttaaa gttacattgg cttccttgaa
     6841 ataaacatgg agtattcaga atgtgtcata aatatttcta attttaagat agtatctcca
     6901 ttggctttct actttttctt ttattttttt ttgtcctctg tcttccattt gttgttgttg
     6961 ttgtttgttt gtttgtttgt tggttggttg gttaattttt ttttaaagat cctacactat
     7021 agttcaagct agactattag ctactctgta acccagggtg accttgaagt catgggtagc
     7081 ctgctgtttt agccttccca catctaagat tacaggtatg agctatcatt tttggtatat
     7141 tgattgattg attgattgat gtgtgtgtgt gtgattgtgt ttgtgtgtgt gattgtgtat
     7201 atgtgtgtat ggttgtgtgt gattgtgtgt atgtatgttt gtgtgtgatt gtgtgtgtgt
     7261 gattgtgcat gtgtgtgtgt gtgattgtgt ttatgtgtat gattgtgtgt gtgtgtgtgt
     7321 gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt tgtgtatata tatttatggt agtgagaggc
     7381 aacgctccgg ctcaggtgtc aggttggttt ttgagacaga gtctttcact tagcttggaa
     7441 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     7501 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     7561 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct
     7621 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     7681 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     7741 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     7801 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     7861 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     7921 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     7981 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     8041 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     8101 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     8161 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     8221 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     8281 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     8341 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     8401 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     8461 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     8521 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     8581 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     8641 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     8701 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     8761 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     8821 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     8881 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     8941 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     9001 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     9061 aggatcttc
//
