LOCUS       sequence-173879-        7267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retroviral stable expression of truncation protein Flag-TZAPZNF1-8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7267)
  AUTHORS   Li JS, Miralles Fuste J, Simavorian T, Bartocci C, Tsai J, Karlseder
            J, Lazzerini Denchi E
  TITLE     TZAP: A telomere-associated protein involved in telomere length 
            control.
  JOURNAL   Science. 2017 Jan 12. pii: aah6752. doi: 10.1126/science.aah6752.
  PUBMED    28082411
REFERENCE   2  (bases 1 to 7267)
  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"; journalName: "Science. 
            2017 Jan 12. pii: aah6752. doi: 10.1126/science.aah6752."
COMMENT     Sequence Label: pLPC-Flag-TZAPZNF1-8
FEATURES             Location/Qualifiers
     source          1..7267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     373..393
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(565..584)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        756..1135
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1136..1339
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1290..1310
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1384..1403
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1384..1402
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1434..1443
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1443..1466
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     LTR             3262..3731
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3861..3878)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4032..4620)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4112..4131)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4780..5571,5572..5640))
                     /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     5403..5422
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5641..5745)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5776..6245
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6310..6509
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6710..7126
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7056..7078
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7094..7110
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             join(7144..7267,1..476)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7144..7163)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
       61 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
      121 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
      181 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
      241 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
      301 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
      361 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
      421 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc
      481 cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca tcggatctcc
      541 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
      601 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
      661 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
      721 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
      781 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      841 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      901 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      961 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     1021 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     1081 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     1141 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     1201 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     1261 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     1321 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
     1381 aattaatacg actcactata gggagaccca agcttggtac cgagctcgga tcagccacca
     1441 tggactacaa agacgatgac gataaaggat catccatgga cggctccttc gtccagcaca
     1501 gtgtgagggt tctgcaggag ctcaacaagc agcgggagaa gggccagtac tgcgacgcca
     1561 ctctggacgt ggggggcctg gtgtttaagg cacactggag tgtccttgcc tgctgcagtc
     1621 actttttcca gagcctctac ggggatggct cagggggcag tgtcgtcctc cctgctggct
     1681 tcgctgagat ctttggcctc ttgttggact ttttctacac tggtcacctc gctctcacct
     1741 cagggaaccg ggatcaggtg ctcctggcag ccagggagtt gcgagtgcca gaggccgtag
     1801 agctgtgcca gagcttcaag cccaaaactt cagtgggaca ggcagcaggt ggccagagtg
     1861 ggctggggcc ccctgcctcc cagaatgtga acagccacgt caaggagccg gcaggcttgg
     1921 aagaagagga agtttcgagg actctgggtc tagtccccag ggatcaggag cccagaggca
     1981 gtcatagtcc tcagaggccc cagctccatt ccccagctca gagtgagggc ccctcctccc
     2041 tctgtgggaa actgaagcag gccttgaagc cttgtcccct tgaggacaag aaacccgagg
     2101 actgcaaagt gcccccaagg cccttagagg ctgaaggtgc ccagctgcag ggcggcagta
     2161 atgagtggga agtggtggtt caagtggagg atgatgggga tggcgattac atgtctgagc
     2221 ctgaggctgt gctgaccagg aggaagtcaa atgtaatccg aaagccctgt gcagctgagc
     2281 cagccctgag cgcgggctcc ctagcagctg agcctgctga gaacagaaaa ggtacagcgg
     2341 tgccggtcga atgccccaca tgtcataaaa agttcctcag caaatattat ctaaaagtcc
     2401 acaacaggaa acatactggg gagaaaccct ttgagtgtcc caaatgtggg aagtgttact
     2461 ttcggaagga gaacctcctg gagcatgaag cccggaattg catgaaccgc tcggaacagg
     2521 tcttcacgtg ctctgtgtgc caggagacat tccgccgaag gatggagctg cgggtgcaca
     2581 tggtgtctca cacaggggag atgccctaca agtgttcctc ctgctcccag cagttcatgc
     2641 agaagaagga cttgcagagc cacatgatca aacttcatgg agcccccaag ccccatgcat
     2701 gccccacctg tgccaagtgc ttcctgtctc ggacagagct gcagctgcat gaagctttca
     2761 agcaccgtgg tgagaagctg tttgtgtgtg aggagtgtgg gcaccgggcc tcgagccgga
     2821 atggcctgca gatgcacatc aaggccaagc acaggaatga gaggccacac gtatgtgagt
     2881 tctgcagcca cgccttcacc caaaaggcca atctcaacat gcacctgcgc acacacacgg
     2941 gtgagaagcc cttccagtgc cacctctgtg gcaagacctt ccgaacccaa gccagcctgg
     3001 acaagcacaa ccgcacccac accggggaaa ggcccttcta ggaattctgc agatatccat
     3061 cacactggcg gccgctcgag catgcatcta gagggccgat aaaataaaag attttattta
     3121 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
     3181 taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag ttcagatcaa
     3241 ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
     3301 cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca aacaggatat
     3361 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt
     3421 ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga
     3481 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
     3541 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc gccagtcctc
     3601 cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg
     3661 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg
     3721 ggggtctttc acatgcagca tgtatcaaaa ttaatttggt tttttttctt aagtatttac
     3781 attaaatggc catagttgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3841 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3901 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3961 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4021 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4081 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4141 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4201 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4261 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4321 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4381 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4441 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4501 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4561 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4621 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4681 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt gcggccgcaa
     4741 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4801 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4861 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4921 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4981 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5041 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5101 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5161 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5221 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5281 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5341 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5401 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5461 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5521 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5581 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5641 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5701 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5761 agtgccacct gcagcctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     5821 ccggctcagg gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg
     5881 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc
     5941 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     6001 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     6061 ctccccgagc tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg
     6121 actgagtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt
     6181 ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc
     6241 tttcatttgg gggctcgtcc gggatcggga gacccctgcc cagggaccac cgacccacca
     6301 ccgggaggca agctggccag caacttatct gtgtctgtcc gattgtctag tgtctatgac
     6361 tgattttatg cgcctgcgtc ggtactagtt agctaactag ctctgtatct ggcggacccg
     6421 tggtggaact gacgagttct gaacacccgg ccgcaaccct gggagacgtc ccagggactt
     6481 tgggggccgt ttttgtggcc cgacctgagg aagggagtcg atgtggaatc cgaccccgtc
     6541 aggatatgtg gttctggtag gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt
     6601 tttgctttcg gtttggaacc gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt
     6661 tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct gaaaattagg gccagactgt
     6721 taccactccc ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa
     6781 ccagtcggta gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac
     6841 ctttaacgtc ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa
     6901 gatcaaggtc ttttcacctg gcccgcatgg acacccagac caggtcccct acatcgtgac
     6961 ctgggaagcc ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc
     7021 tccgcctcct cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc
     7081 gcctcgatcc tccctttatc cagccctcac tccttctcta ggcgccggcc ggatcagctt
     7141 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     7201 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     7261 gaccgcc
//
