LOCUS       sequence-288028-        6960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of 3xFLAG tagged pontin, N-terminal..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6960)
  AUTHORS   Venteicher AS, Meng Z, Mason PJ, Veenstra TD, Artandi SE
  TITLE     Identification of ATPases pontin and reptin as telomerase components
            essential for holoenzyme assembly.
  JOURNAL   Cell. 2008 Mar 21;132(6):945-57. doi: 10.1016/j.cell.2008.01.019.
  PUBMED    18358808
REFERENCE   2  (bases 1 to 6960)
  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.cell.2008.01"; journalName: "Cell"; date: "2008-03-21- 
            21"; volume: "132"; issue: "6"; pages: "945-57"
COMMENT     Sequence Label: pCDNA-3xFLAG-Pontin
FEATURES             Location/Qualifiers
     source          1..6960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      726..735
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             747..812
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2230..2259
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2275..2292
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2315..2332)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2321..2545
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2591..3019
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2678..2697)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2888..2909
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3028..3048)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3033..3362
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3213..3348
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3275..3294
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3429..4223
                     /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(3483..3502)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4093..4112
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4399..4520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4436..4455)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4490..4509
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4569..4585)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4569..4585)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4582..4604)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4593..4609
                     /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(4617..4623,4624..4641,4642..4647))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4662..4683
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4800..4817)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4971..5559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5051..5070)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5730..6521,6522..6590))
                     /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     6353..6372
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6591..6695)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6770..6789)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga attctgcaga
      721 taaacgccac catggggtcc cgggctgact acaaagacca tgacggtgat tataaagatc
      781 atgacatcga ctacaaggat gacgatgaca aggggcgcgc caagattgag gaggtgaaga
      841 gcactacgaa gacgcagcgc atcgcctccc acagccacgt gaaagggctg gggctggacg
      901 agagcggctt ggccaagcag gcggcctcag ggcttgtggg ccaggagaac gcgcgagagg
      961 catgtggcgt catagtagaa ttaatcaaaa gcaagaaaat ggctggaaga gctgtcttgt
     1021 tggcaggacc tcctggaact ggcaagacag ctctggctct ggctattgct caggagctgg
     1081 gtagtaaggt ccccttctgc ccaatggtgg ggagtgaagt ttactcaact gagatcaaga
     1141 agacagaggt gctgatggag aacttccgca gggccattgg gctgcgaata aaggagacca
     1201 aggaagttta tgaaggtgaa gtcacagagc taactccgtg tgagacagag aatcccatgg
     1261 gaggatatgg caaaaccatt agccatgtga tcataggact caaaacagcc aaaggaacca
     1321 aacagttgaa actggacccc agcatttttg aaagtttgca gaaagagcga gtagaagctg
     1381 gagatgtgat ttacattgaa gccaacagtg gggccgtgaa gaggcagggc aggtgtgata
     1441 cctatgccac agaattcgac cttgaagctg aagagtatgt ccccttgcca aaaggggatg
     1501 tgcacaaaaa gaaagaaatc atccaagatg tgaccttgca tgacttggat gtggctaatg
     1561 cgcggcccca ggggggacaa gatatcctgt ccatgatggg ccagctaatg aagccaaaga
     1621 agacagaaat cacagacaaa cttcgagggg agattaataa ggtggtgaac aagtacatcg
     1681 accagggcat tgctgagctg gtcccgggtg tgctgtttgt tgatgaggtc cacatgctgg
     1741 acattgagtg cttcacctac ctgcaccgcg ccctggagtc ttctatcgct cccatcgtca
     1801 tctttgcatc caaccgaggc aactgtgtca tcagaggcac tgaggacatc acatcccctc
     1861 acggcatccc tcttgacctt ctggaccgag tgatgataat ccggaccatg ctgtatactc
     1921 cacaggaaat gaaacagatc attaaaatcc gtgcccagac ggaaggaatc aacatcagtg
     1981 aggaggcact gaaccacctg ggggagattg gcaccaagac cacactgagg tactcagtgc
     2041 agctgctgac cccggccaac ttgcttgcta aaatcaacgg gaaggacagc attgagaaag
     2101 agcatgtcga agagatcagt gaacttttct atgatgccaa gtcctccgcc aaaatcctgg
     2161 ctgaccagca ggataagtac atgaagtgat taattaagtc tagagtcgag tctagagggc
     2221 ccgcggttcg aacaaaaact catctcagaa gaggatctga atatgcatac cggtcatcat
     2281 caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     2341 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2401 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2461 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     2521 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     2581 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2641 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2701 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2761 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2821 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2881 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2941 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3001 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3061 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3121 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3181 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3241 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3301 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3361 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     3421 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3481 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3541 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3601 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3661 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3721 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3781 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3841 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3901 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     3961 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4021 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4081 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4141 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4201 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg
     4261 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4321 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4381 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4441 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4501 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4561 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4621 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4681 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4741 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4801 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4861 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4921 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4981 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5041 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5101 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5161 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5221 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5281 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5341 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5401 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5461 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5521 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5581 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5641 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5701 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5761 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5821 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5881 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5941 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6001 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6061 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6121 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6181 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6241 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6301 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6361 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6421 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6481 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6541 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6601 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6661 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6721 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     6781 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     6841 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     6901 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
//
