LOCUS       sequence-172953-        6522 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Retroviral vector expressing RAP2A-GFP-IRES-Bsd.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6522)
  AUTHORS   Wang T, Yu H, Hughes NW, Liu B, Kendirli A, Klein K, Chen WW, Lander
            ES, Sabatini DM
  TITLE     Gene Essentiality Profiling Reveals Gene Networks and Synthetic 
            Lethal Interactions with Oncogenic Ras.
  JOURNAL   Cell. 2017 Feb 1. pii: S0092-8674(17)30061-2. doi: 
            10.1016/j.cell.2017.01.013.
  PUBMED    28162770
REFERENCE   2  (bases 1 to 6522)
  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: "Cell. 2017 
            Feb 1. pii: S0092-8674(17)30061-2. doi: 10.1016/j.cell.2017.01.013."
COMMENT     Sequence Label: pMXs-RAP2A-GFP
FEATURES             Location/Qualifiers
     source          1..6522
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             153..548
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             719..1188
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(1318..1335)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1489..2077)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1569..1588)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2241..3032,3033..3101))
                     /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     2864..2883
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3102..3206)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             3236..3829
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    3892..4249
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             4308..4724
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4654..4676
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4692..4708
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    4734..5108
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     5046..5062
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             join(5716..5718,5719..5721,5722..6432)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5716..5718,5719..5721,5722..6432)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5761..5782)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6022..6041)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6369..6390
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6445..6510
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
       61 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
      121 ttgaaaaaca cgatgataat atggccacaa ccatggttcc tttgtctcaa gaagaatcca
      181 ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg
      241 tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt
      301 ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg
      361 gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg
      421 gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca
      481 gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg
      541 agggctaagt cgacgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg
      601 aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg
      661 aaaaatacat aactgagaat agagaagttc agatcaaggt caggaacaga tggaacagct
      721 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      781 acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
      841 ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag
      901 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
      961 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
     1021 aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta
     1081 cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg
     1141 ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcaca tgcagcatgt
     1201 atcaaaatta atttggtttt ttttcttaag tatttacatt aaatggccat agttgcatta
     1261 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     1321 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1381 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1441 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1501 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1561 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1621 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1681 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1741 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1801 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1861 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1921 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1981 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2041 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2101 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2161 aaaaaggatc ttcacctaga tccttttgcg gccggccgca aatcaatcta aagtatatat
     2221 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2281 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2341 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     2401 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2461 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2521 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     2581 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     2641 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     2701 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     2761 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2821 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     2881 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2941 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3001 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3061 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3121 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3181 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgcataatga
     3241 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     3301 aaaatacata actgagaata gaaaagttca gatcaaggtc aggaacagat ggaacagctg
     3361 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     3421 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     3481 gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga
     3541 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     3601 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     3661 agagcccaca acccctcact cggcgcgcca gtcctccgat tgactgagtc gcccgggtac
     3721 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     3781 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt
     3841 ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
     3901 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta tgcgcctgcg
     3961 tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
     4021 cggaacaccc ggccgcaacc ctgggagacg tcccagggac ttcgggggcc gtttttgtgg
     4081 cccgacctga gtccaaaaat cccgatcgtt ttggactctt tggtgcaccc ccctaatagg
     4141 agggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat
     4201 ttttgctttc ggtttgggac cgaagccgcg ccgcgcgtct tgtctgctgc agcatcgttc
     4261 tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta
     4321 ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc
     4381 agtcggtaga tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct
     4441 ttaacgtcgg atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga
     4501 tcaaggtctt ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct
     4561 gggaagcctt ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc
     4621 cgcctcctct tcctccatcc gccccgtctc tcccccttga acctcctcgt tcgaccccgc
     4681 ctcgatcctc cctttatcca gccctcactc cttctctagg cgcccccata tggccatatg
     4741 agatcttata tggggcaccc ccgccccttg taaacttccc tgaccctgac atgacaagag
     4801 ttactaacag cccctctctc caagctcact tacaggctct ctacttagtc cagcacgaag
     4861 tctggagacc tctggcggca gcctaccaag aacaactgga ccgaccggtg gtacctcacc
     4921 cttaccgagt cggcgacaca gtgtgggtcc gccgacacca gactaagaac ctagaacctc
     4981 gctggaaagg accttacaca gtcctgctga ccacccccac cgccctcaaa gtagacggca
     5041 tcgcagcttg gatacacgcc gcccacgtga aggctgccga ccccgggggt ggaccatcct
     5101 ctagactgcc ggatctagct agttaattaa ggatccgcca ccatgcgcga gtacaaagtg
     5161 gtggtgctgg gctcgggcgg ggtaggcaaa tccgccctga ccgtgcagtt cgtgaccggc
     5221 accttcatcg agaaatacga ccccaccatc gaggacttct accgcaagga gatcgaggtg
     5281 gattcgtcgc cgtcggtgct ggagatcctg gacacggcgg gcaccgagca gttcgcgtcc
     5341 atgcgggacc tgtacatcaa gaacggccag ggcttcatcc tcgtctacag cctcgtcaac
     5401 cagcagagct tccaggacat caagcccatg cgggaccaga tcatccgcgt gaagcggtat
     5461 gagaaagtgc cagtcatctt ggttgggaac aaagtggacc tggaaagtga gagagaagta
     5521 tcgtccagcg aaggcagagc ccttgctgaa gagtggggct gcccctttat ggaaacttcc
     5581 gctaagagta aaacaatggt ggacgaactc tttgcagaaa ttgtgaggca gatgaactat
     5641 gctgctcagc ctgacaaaga tgacccatgc tgttctgcat gtaacataca aggaggcggt
     5701 agcgcggccg caggcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     5761 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     5821 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     5881 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     5941 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     6001 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     6061 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     6121 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     6181 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     6241 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     6301 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     6361 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     6421 gagctgtaca agggcggagg kggcgactac aaagaccatg acggtgatta taaagatcat
     6481 gacattgatt acaaggatga cgatgacaag taataataaa tt
//
