LOCUS       sequence-362645-        6938 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expressions of mouse RAP1 I312R.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6938)
  AUTHORS   Barry RM, Sacco O, Mameri A, Stojaspal M, Kartsonis W, Shah P, De 
            Ioannes P, Hofr C, Cote J, Sfeir A
  TITLE     Rap1 regulates TIP60 function during fate transition between 
            two-cell-like and pluripotent states.
  JOURNAL   Genes Dev. 2022 Feb 24. pii: gad.349039.121. doi: 
            10.1101/gad.349039.121.
  PUBMED    35210222
REFERENCE   2  (bases 1 to 6938)
  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: "Genes Dev. 
            2022 Feb 24. pii: gad.349039.121. doi: 10.1101/gad.349039.121."
COMMENT     Sequence Label: pLPC-FLAG.HA.HA.Mm.RAP1.I312R
FEATURES             Location/Qualifiers
     source          1..6938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             2..471
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    536..735
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             936..1352
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1282..1304
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1320..1336
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1370..1969
                     /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(1370..1389)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1866..1886
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2058..2077)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2249..2628
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2629..2832
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2783..2803
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2877..2896
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2877..2895
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2908..2917
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2917..2940
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2953..2979
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             2953..2979
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2989..3015
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             2989..3015
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     LTR             4426..4895
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5025..5042)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5196..5784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5276..5295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5944..6735,6736..6804))
                     /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     6567..6586
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6805..6909)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
       61 agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
      121 cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta
      181 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
      241 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
      301 taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg
      361 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc
      421 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc atttgggggc
      481 tcgtccggga tcgggagacc cctgcccagg gaccaccgac ccaccaccgg gaggcaagct
      541 ggccagcaac ttatctgtgt ctgtccgatt gtctagtgtc tatgactgat tttatgcgcc
      601 tgcgtcggta ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg
      661 agttctgaac acccggccgc aaccctggga gacgtcccag ggactttggg ggccgttttt
      721 gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga tatgtggttc
      781 tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt
      841 ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg tgttgtctct
      901 gtctgactgt gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa
      961 gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg
     1021 tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat
     1081 ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt
     1141 cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg
     1201 cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc
     1261 ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc
     1321 tttatccagc cctcactcct tctctaggcg ccggccggat cagcttacca tgaccgagta
     1381 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     1441 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     1501 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     1561 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     1621 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     1681 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     1741 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     1801 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     1861 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     1921 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca
     1981 cgacccgcag cgcccgaccg aaaggagcgc acgaccccat gcatcggatc tcccgatccc
     2041 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     2101 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     2161 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     2221 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
     2281 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     2341 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     2401 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     2461 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     2521 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     2581 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     2641 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     2701 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     2761 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     2821 ataagcagag ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat
     2881 acgactcact atagggagac ccaagctgcc accatggact acaaggacga cgatgacaag
     2941 ctcgatggag gataccccta cgacgtgccc gactacgccg ctggagcata cccctacgac
     3001 gtgcccgact acgccctcga ggcaggatcc gcggaggcga tggatctggg taaagacccc
     3061 aatgggccca ctcactcctc cactctgttc gtgagagaag acggcagcgc catgtcgttt
     3121 tacgtgcggc ccagctcggc caagcgccgg ctgtcgacgc tcatcctgca cggcggcggc
     3181 accgtgtgtc gggtgcagga acccggagcc gtgcttctcg cccagcccgg ggaggcgctg
     3241 gccgaggctt cgggcgactt catctccacg cagtacatcc tagactgcgt ggatcgcaac
     3301 gagaagctgg acctggaggc ctatcggctg ggcctgacgg agcaggcgtc cgatccgaag
     3361 cccggggctt ccaccgaggg ctccacggaa ccggagccgc agcccctgac cgggcgcatc
     3421 gcctacaccg acgcggaaga tgtggccatc ctgacctacg tgaaggagaa cgcccgttcg
     3481 cccagctcgg tcacaggcaa tgccttgtgg aaagcgatgg agaagagctc gctcacgcag
     3541 cactcctggc agtcgctcaa ggaccgctac ctcaagcacc tacggggtca ggagcacaag
     3601 tacctgctcg ggaacgcccc ggtcagcccg tcctcccaga agctcaaacg gaaggcggag
     3661 caggaccccg aggccgcgga tagcggagag ccacagaaca agagagcgcc agacttgcct
     3721 gaggaggagt gtgtgaaagg agagatcaag gagaatggag aggcagacaa caagctgttt
     3781 gaggaagccg ctccggagtt cggagaagcc gtggtggatg agagccctga ctttgaaata
     3841 catataacga tgtgtgatgg tgatccaccc acacccgagg aagactcaga aacacagcca
     3901 gacgaggagg aagaagaacc aaaagtttct acgcaagaag tgggaactgc cattaaggtg
     3961 cgccggcagc taatggagaa gttcaacttg gatctatcaa cagttacaca ggccttgctg
     4021 aagaacagtg gtgagctgga ggccacgtcc tccttcttag agtcggggcg gagacccgac
     4081 ggttatccca tttggtgcag acaagatgac ttagatttgc agaaggacga tgacgacacg
     4141 aaaaatgcac tggtcaaaaa gtttggagct cagaacgttg ctcggaggat tgaattccga
     4201 aagaaataaa agcttaggtg gaattctctg actagctgat cgagcatgca tctagagggc
     4261 cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     4321 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac
     4381 tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa
     4441 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag
     4501 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     4561 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
     4621 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     4681 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     4741 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
     4801 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     4861 tgagtgattg actacccgtc agcgggggtc tttcacatgc agcatgtatc aaaattaatt
     4921 tggttttttt tcttaagtat ttacattaaa tggccatagt tgcattaatg aatcggccaa
     4981 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5041 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5101 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5161 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5221 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5281 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5341 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5401 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5461 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5521 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5581 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5641 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5701 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5761 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5821 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5881 acctagatcc ttttgcggcc gcaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5941 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6001 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6061 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6121 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6181 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6241 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6301 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6361 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6421 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6481 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6541 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6601 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6661 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6721 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6781 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6841 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6901 ggttccgcgc acatttcccc gaaaagtgcc acctgcag
//
