LOCUS       sequence-173803-        7765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retroviral stable expression of Flag-TZAP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7765)
  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 7765)
  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-TZAP
FEATURES             Location/Qualifiers
     source          1..7765
                     /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             3760..4229
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4359..4376)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4530..5118)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4610..4629)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5278..6069,6070..6138))
                     /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     5901..5920
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6139..6243)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             6274..6743
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6808..7007
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             7208..7624
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7554..7576
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7592..7608
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             join(7642..7765,1..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(7642..7661)
                     /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 ccatggacgg ctccttcgtc cagcacagtg
     1501 tgagggttct gcaggagctc aacaagcagc gggagaaggg ccagtactgc gacgccactc
     1561 tggacgtggg gggcctggtg tttaaggcac actggagtgt ccttgcctgc tgcagtcact
     1621 ttttccagag cctctacggg gatggctcag ggggcagtgt cgtcctccct gctggcttcg
     1681 ctgagatctt tggcctcttg ttggactttt tctacactgg tcacctcgct ctcacctcag
     1741 ggaaccggga tcaggtgctc ctggcagcca gggagttgcg agtgccagag gccgtagagc
     1801 tgtgccagag cttcaagccc aaaacttcag tgggacaggc agcaggtggc cagagtgggc
     1861 tggggccccc tgcctcccag aatgtgaaca gccacgtcaa ggagccggca ggcttggaag
     1921 aagaggaagt ttcgaggact ctgggtctag tccccaggga tcaggagccc agaggcagtc
     1981 atagtcctca gaggccccag ctccattccc cagctcagag tgagggcccc tcctccctct
     2041 gtgggaaact gaagcaggcc ttgaagcctt gtccccttga ggacaagaaa cccgaggact
     2101 gcaaagtgcc cccaaggccc ttagaggctg aaggtgccca gctgcagggc ggcagtaatg
     2161 agtgggaagt ggtggttcaa gtggaggatg atggggatgg cgattacatg tctgagcctg
     2221 aggctgtgct gaccaggagg aagtcaaatg taatccgaaa gccctgtgca gctgagccag
     2281 ccctgagcgc gggctcccta gcagctgagc ctgctgagaa cagaaaaggt acagcggtgc
     2341 cggtcgaatg ccccacatgt cataaaaagt tcctcagcaa atattatcta aaagtccaca
     2401 acaggaaaca tactggggag aaaccctttg agtgtcccaa atgtgggaag tgttactttc
     2461 ggaaggagaa cctcctggag catgaagccc ggaattgcat gaaccgctcg gaacaggtct
     2521 tcacgtgctc tgtgtgccag gagacattcc gccgaaggat ggagctgcgg gtgcacatgg
     2581 tgtctcacac aggggagatg ccctacaagt gttcctcctg ctcccagcag ttcatgcaga
     2641 agaaggactt gcagagccac atgatcaaac ttcatggagc ccccaagccc catgcatgcc
     2701 ccacctgtgc caagtgcttc ctgtctcgga cagagctgca gctgcatgaa gctttcaagc
     2761 accgtggtga gaagctgttt gtgtgtgagg agtgtgggca ccgggcctcg agccggaatg
     2821 gcctgcagat gcacatcaag gccaagcaca ggaatgagag gccacacgta tgtgagttct
     2881 gcagccacgc cttcacccaa aaggccaatc tcaacatgca cctgcgcaca cacacgggtg
     2941 agaagccctt ccagtgccac ctctgtggca agaccttccg aacccaagcc agcctggaca
     3001 agcacaaccg cacccacacc ggggaaaggc ccttcagttg cgagttctgt gaacagcgct
     3061 tcactgagaa ggggcccctc ctgaggcacg tggccagccg ccatcaggag ggccggcccc
     3121 acttctgcca gatatgcggc aagaccttca aagccgtgga gcaactgcgt gtgcacgtca
     3181 gacggcacaa gggggtgagg aagtttgagt gcaccgagtg tggctacaag tttacccgac
     3241 aggcccacct gcggaggcac atggagatcc acgaccgggt agagaactac aacccgcggc
     3301 agcgcaagct ccgcaacctg atcatcgagg acgagaagat ggtggtggtg gcgctgcagc
     3361 cgcctgcaga gctggaggtg ggctcggcgg aggtcattgt ggagtccctg gcccagggcg
     3421 gcctggcctc ccagctcccc ggccagagac tgtgtgcaga ggagagcttc accggcccag
     3481 gtgtcctgga gccctccctc atcatcacag ctgctgtccc cgaggactgt gacacatagg
     3541 aattctgcag atatccatca cactggcggc cgctcgagca tgcatctaga gggccgataa
     3601 aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt
     3661 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaaataca taactgagaa
     3721 tagagaagtt cagatcaagg tcaggaacag atggaacagc tgaatatggg ccaaacagga
     3781 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatgga acagctgaat
     3841 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     3901 atggtcccca gatgcggtcc agccctcagc agtttctaga gaaccatcag atgtttccag
     3961 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
     4021 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
     4081 ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc
     4141 ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg
     4201 attgactacc cgtcagcggg ggtctttcac atgcagcatg tatcaaaatt aatttggttt
     4261 tttttcttaa gtatttacat taaatggcca tagttgcatt aatgaatcgg ccaacgcgcg
     4321 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4381 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4441 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4501 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4561 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4621 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4681 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4741 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4801 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4861 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4921 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4981 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5041 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5101 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5161 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5221 atccttttgc ggccgcaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5281 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5341 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5401 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5461 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5521 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5581 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5641 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5701 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5761 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5821 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5881 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5941 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6001 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6061 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6121 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6181 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6241 gcgcacattt ccccgaaaag tgccacctgc agcctgaata tgggccaaac aggatatctg
     6301 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc
     6361 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc
     6421 cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc
     6481 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct
     6541 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg
     6601 gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg
     6661 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
     6721 tacccgtcag cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca
     6781 gggaccaccg acccaccacc gggaggcaag ctggccagca acttatctgt gtctgtccga
     6841 ttgtctagtg tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct
     6901 ctgtatctgg cggacccgtg gtggaactga cgagttctga acacccggcc gcaaccctgg
     6961 gagacgtccc agggactttg ggggccgttt ttgtggcccg acctgaggaa gggagtcgat
     7021 gtggaatccg accccgtcag gatatgtggt tctggtagga gacgagaacc taaaacagtt
     7081 cccgcctccg tctgaatttt tgctttcggt ttggaaccga agccgcgcgt cttgtctgct
     7141 gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga
     7201 aaattagggc cagactgtta ccactccctt aagtttgacc ttaggtcact ggaaagatgt
     7261 cgagcggatc gctcacaacc agtcggtaga tgtcaagaag agacgttggg ttaccttctg
     7321 ctctgcagaa tggccaacct ttaacgtcgg atggccgcga gacggcacct ttaaccgaga
     7381 cctcatcacc caggttaaga tcaaggtctt ttcacctggc ccgcatggac acccagacca
     7441 ggtcccctac atcgtgacct gggaagcctt ggcttttgac ccccctccct gggtcaagcc
     7501 ctttgtacac cctaagcctc cgcctcctct tcctccatcc gccccgtctc tcccccttga
     7561 acctcctcgt tcgaccccgc ctcgatcctc cctttatcca gccctcactc cttctctagg
     7621 cgccggccgg atcagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc
     7681 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     7741 cgccacaccg tcgatccgga ccgcc
//
