LOCUS       sequence-362572-        6523 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of mouse Epc1 EPcA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6523)
  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 6523)
  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.Mm.Epc1-EPcA
FEATURES             Location/Qualifiers
     source          1..6523
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(930..1721,1722..1790))
                     /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     1553..1572
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1791..1895)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             1926..2395
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    2460..2659
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             2860..3276
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3206..3228
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3244..3260
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             3294..3893
                     /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(3294..3313)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3790..3810
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3982..4001)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4173..4552
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4553..4756
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4707..4727
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4801..4820
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4801..4819
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4851..4860
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4860..4883
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     LTR             5935..6404
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt gcggccgcaa atcaatctaa
      901 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
      961 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     1021 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     1081 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     1141 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     1201 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     1261 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     1321 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1381 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1441 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     1501 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     1561 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     1621 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     1681 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     1741 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     1801 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     1861 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     1921 gcagcctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     1981 gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
     2041 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt
     2101 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
     2161 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
     2221 tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc
     2281 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg
     2341 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg
     2401 gggctcgtcc gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggca
     2461 agctggccag caacttatct gtgtctgtcc gattgtctag tgtctatgac tgattttatg
     2521 cgcctgcgtc ggtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact
     2581 gacgagttct gaacacccgg ccgcaaccct gggagacgtc ccagggactt tgggggccgt
     2641 ttttgtggcc cgacctgagg aagggagtcg atgtggaatc cgaccccgtc aggatatgtg
     2701 gttctggtag gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg
     2761 gtttggaacc gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt
     2821 ctctgtctga ctgtgtttct gtatttgtct gaaaattagg gccagactgt taccactccc
     2881 ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta
     2941 gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc
     3001 ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc
     3061 ttttcacctg gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc
     3121 ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct
     3181 cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc
     3241 tccctttatc cagccctcac tccttctcta ggcgccggcc ggatcagctt accatgaccg
     3301 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     3361 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     3421 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     3481 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     3541 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     3601 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     3661 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     3721 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     3781 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     3841 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc
     3901 cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg gatctcccga
     3961 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     4021 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     4081 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     4141 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
     4201 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     4261 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     4321 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     4381 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     4441 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     4501 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     4561 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     4621 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     4681 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     4741 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
     4801 taatacgact cactataggg agacccaagc ttggtaccga gctcggatca gccaccatgg
     4861 actacaaaga cgatgacgat aaaggatcca gtaaactgtc gtttcgggcg cgggcgctgg
     4921 acgcctcgaa gccgctgccg gtcttccgct gtgaggatct gcccgacctg cacgaatacg
     4981 cctcgataaa ccgggccgtg ccgcagatgc ccaccgggat ggagaaggaa gaggagtcgg
     5041 aacaccatct tcagcgggct atttctgcac agcaggtgta cggcgagaag cgggataaca
     5101 tggttattcc agttcctgaa gcagagagta acatcgctta ctatgagtcc atataccctg
     5161 gggagttcag gatgccaaag cagctcattc acatacagcc ttttagcttg gacgctgaac
     5221 agcctgatta cgatctggat tctgaagatg aagtatttgt gaataaactg aagaagaaaa
     5281 tggacatctg cccattgcag tttgaggaga tgattgaccg gctagaaaaa ggcagcggtc
     5341 agcagccagt cagtctgcag gaagccaaac tacttctaaa agaagatgat gagttaatta
     5401 gagaagttta tgaatactgg attaaaaaga ggaaaacctg ccgggggtca tcacttatcc
     5461 cgttggtaaa acaggaaaag cgagatggtt ctagcacaaa tgatccttac gtggctttta
     5521 gaagacgaac tgagaaaatg caaactcgga aaaaccgtaa gaatgatgaa gcctcatatg
     5581 agaaaatgct caagctgcgc cgcgatctca gccgggctgt caccatcctg gaaatgatca
     5641 agagacggga gaagagtaaa cgggagctgc tgcacctgac cctggaaatc atggagaaga
     5701 ggtataattt ataagaattc tgcagatatc catcacactg gcggccgctc gagcatgcat
     5761 ctagagggcc gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag
     5821 accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa
     5881 atacataact gagaatagag aagttcagat caaggtcagg aacagatgga acagctgaat
     5941 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     6001 atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     6061 cagggccaag aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagagaacc
     6121 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     6181 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga
     6241 gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg
     6301 tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag
     6361 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcacatgca gcatgtatca
     6421 aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagtt gcattaatga
     6481 atcggccaac gcgcggggag aggcggtttg cgtattgggc gct
//
