LOCUS       sequence-372393-        8167 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of BirA-mRap1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8167)
  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 8167)
  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-BirA-Linker-Myc-mRap1
FEATURES             Location/Qualifiers
     source          1..8167
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             942..1358
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1288..1310
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1326..1342
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1376..1975
                     /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(1376..1395)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1872..1892
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2064..2083)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2255..2634
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2635..2838
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2789..2809
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2883..2902
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2883..2901
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2933..2942
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2942..2965
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(2972..3319,3320..3322,3323..3931)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             4139..4168
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(6260..6277)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6431..7019)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6511..6530)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7179..7970,7971..8039))
                     /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     7802..7821
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8040..8144)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
      901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
      961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatcagc ttaccatgac
     1381 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     1441 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     1501 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     1561 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     1621 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     1681 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     1741 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     1801 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     1861 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     1921 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacgccc
     1981 gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat cggatctccc
     2041 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     2101 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     2161 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     2221 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
     2281 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     2341 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     2401 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     2461 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     2521 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     2581 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     2641 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     2701 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     2761 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     2821 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
     2881 attaatacga ctcactatag ggagacccaa gcttggtacc gagctcggat cagccaccat
     2941 ggactacaaa gacgatgacg ataaaggatc caaggacaac accgtgcccc tgaagctgat
     3001 cgccctgctg gccaatggcg agttccactc tggcgagcag ctgggcgaga cactgggcat
     3061 gtctagagcc gccatcaaca agcacatcca gaccctgaga gactggggcg tggacgtgtt
     3121 caccgtgcct ggcaagggct acagcctgcc cgagcctatc cagctgctga acgccaagca
     3181 gatcctgggc cagctggatg gcggatctgt ggctgtgctg cccgtgatcg acagcaccaa
     3241 ccagtacctg ctggaccgga tcggcgagct gaaaagcggc gacgcctgta tcgccgagta
     3301 tcagcaggct ggcagaggcg gcaggggcag aaagtggttc tctcccttcg gcgccaatct
     3361 gtacctgagc atgttctggc ggctggaaca gggacctgcc gccgctattg gactgagcct
     3421 cgtgatcggc atcgtgatgg ccgaggtgct gagaaagctg ggcgccgaca aagtgcgcgt
     3481 gaagtggccc aacgacctgt atctgcagga ccggaagctg gccggcatcc tggtggaact
     3541 gacaggcaaa acaggcgacg ctgcccagat cgtgattggc gctggcatca atatggccat
     3601 gcggcgggtg gaagagtccg tcgtgaatca gggctggatc accctgcagg aagccggcat
     3661 caacctggac agaaacaccc tggccgccat gctgatcaga gagctgagag ccgccctgga
     3721 actgttcgag caggaaggcc tggcccccta cctgtccaga tgggagaagc tggacaactt
     3781 catcaacaga cccgtgaaac tgatcatcgg cgacaaagag atcttcggca tcagccgggg
     3841 catcgacaag cagggcgctc tgctgctgga acaggacggc atcatcaagc cctggatggg
     3901 cggcgagatc agcctgagat ctgccgagaa agaattcgga ggcggtgggt cgggaggtgg
     3961 cggatctgga gggggcgggt caggtggcgg aggttcaggg ggtggaggct ctggcggagg
     4021 gggctccggt ggkgggggat ccggaggcgg tgggtcggga ggtggcggat ctggaggggg
     4081 cgggtcaggt ggcggaggtt cagggggtgg aggctctggc ggagggggct ccctcgagga
     4141 acaaaagttg atttctgaag aagatttgtt aagaaagaga ggatccgcgg aggcgatgga
     4201 tctgggtaaa gaccccaatg ggcccactca ctcctccact ctgttcgtga gagaagacgg
     4261 cagcgccatg tcgttttacg tgcggcccag ctcggccaag cgccggctgt cgacgctcat
     4321 cctgcacggc ggcggcaccg tgtgtcgggt gcaggaaccc ggagccgtgc ttctcgccca
     4381 gcccggggag gcgctggccg aggcttcggg cgacttcatc tccacgcagt acatcctaga
     4441 ctgcgtggat cgcaacgaga agctggacct ggaggcctat cggctgggcc tgacggagca
     4501 ggcgtccgat ccgaagcccg gggcttccac cgagggctcc acggaaccgg agccgcagcc
     4561 cctgaccggg cgcatcgcct acaccgacgc ggaagatgtg gccatcctga cctacgtgaa
     4621 ggagaacgcc cgttcgccca gctcggtcac aggcaatgcc ttgtggaaag cgatggagaa
     4681 gagctcgctc acgcagcact cctggcagtc gctcaaggac cgctacctca agcacctacg
     4741 gggtcaggag cacaagtacc tgctcgggaa cgccccggtc agcccgtcct cccagaagct
     4801 caaacggaag gcggagcagg accccgaggc cgcggatagc ggagagccac agaacaagag
     4861 agcgccagac ttgcctgagg aggagtgtgt gaaaggagag atcaaggaga atggagaggc
     4921 agacaacaag ctgtttgagg aagccgctcc ggagttcgga gaagccgtgg tggatgagag
     4981 ccctgacttt gaaatacata taacgatgtg tgatggtgat ccacccacac ccgaggaaga
     5041 ctcagaaaca cagccagacg aggaggaaga agaaccaaaa gtttctacgc aagaagtggg
     5101 aactgccatt aaggtgatcc ggcagctaat ggagaagttc aacttggatc tatcaacagt
     5161 tacacaggcc ttgctgaaga acagtggtga gctggaggcc acgtcctcct tcttagagtc
     5221 ggggcggaga cccgacggtt atcccatttg gtgcagacaa gatgacttag atttgcagaa
     5281 ggacgatgac gacacgaaaa atgcactggt caaaaagttt ggagctcaga acgttgctcg
     5341 gaggattgaa ttccgaaaga aataagcatg catctagagg gccgataaaa taaaagattt
     5401 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     5461 cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata gagaagttca
     5521 gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     5581 gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat gggccaaaca
     5641 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     5701 tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
     5761 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
     5821 cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact cggggcgcca
     5881 gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagtyg
     5941 cwtcygyccc csrgsycaat aaaagagccc acaacccctc actcggggcg ccagtcctcc
     6001 gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga
     6061 cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg
     6121 gggtctttca catgcagcat gtatcaaaat taatttggtt ttttttctta agtatttaca
     6181 ttaaatggcc atagttgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6241 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6301 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6361 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6421 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6481 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6541 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6601 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6661 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6721 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6781 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6841 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6901 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6961 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7021 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7081 ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg cggccgcaaa
     7141 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7201 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7261 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7321 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7381 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7441 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7501 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7561 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7621 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7681 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7741 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7801 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7861 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7921 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7981 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8041 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8101 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8161 gtgccac
//
