LOCUS       sequence-197151-        7298 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses GFP tagged metap2 in mammalian cells to make lentivirus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7298)
  AUTHORS   Gu X, Orozco JM, Saxton RA, Condon KJ, Liu GY, Krawczyk PA, Scaria 
            SM, Harper JW, Gygi SP, Sabatini DM
  TITLE     SAMTOR is an S-adenosylmethionine sensor for the mTORC1 pathway.
  JOURNAL   Science. 2017 Nov 10;358(6364):813-818. doi: 
            10.1126/science.aao3265.
  PUBMED    29123071
REFERENCE   2  (bases 1 to 7298)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2017-11-10- 10"; volume: "358"; 
            issue: "6364"; pages: "813-818"
COMMENT     Sequence Label: pLC242-GFP-metap2
FEATURES             Location/Qualifiers
     source          1..7298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..470
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    535..734
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             935..1351
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1281..1303
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1319..1335
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1386..1395
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1392..1394,1395..1397,1398..2108)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1392..1394,1395..1397,1398..2108)
                     /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(1437..1458)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1698..1717)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2045..2066
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2154..2174
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2187..2231
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3865..3885)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3870..4199
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4050..4185
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4207..4605
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4774..5243
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5373..5390)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5544..6132)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5624..5643)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6303..7094,7095..7163))
                     /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     6926..6945
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7164..7268)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
       61 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      121 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      181 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      241 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
      301 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
      361 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
      421 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
      481 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggcaagctg
      541 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
      601 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      661 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg
      721 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct
      781 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
      841 gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg
      901 tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag
      961 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt
     1021 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg
     1081 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc
     1141 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc
     1201 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
     1261 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
     1321 ttatccagcc ctcactcctt ctctaggcgc cggccggatc ccagtgtggt ggtacgctac
     1381 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1441 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1501 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     1561 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     1621 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1681 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1741 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1801 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     1861 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     1921 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1981 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     2041 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2101 tgtacaagtc cggaggggat cggtggagct ccaccggtgg cggccgctct agagagaatc
     2161 tttattttca gggcgccgcc aaattcaaag aaaccgctgc tgctaaattc gaacgccagc
     2221 acatggacag cggaggtgga ggtactagtc tcgagatggc gggtgtggag gaggtagcgg
     2281 cctccgggag ccacctgaat ggcgacctgg atccagacga cagggaagaa ggagctgcct
     2341 ctacggctga ggaagcagcc aagaaaaaaa gacgaaagaa gaagaagagc aaagggcctt
     2401 ctgcagggga acaggaacct gataaagaat caggagcctc agtggatgaa gtagcaagac
     2461 agttggaaag atcagcattg gaagataaag aaagagatga agatgatgaa gatggagatg
     2521 gcgatggaga tggagcaact ggaaagaaga agaaaaagaa gaagaagaag agaggaccaa
     2581 aagttcaaac agaccctccc tcagttccaa tatgtgacct gtatcctaat ggtgtatttc
     2641 ccaaaggaca agaatgcgaa tacccaccca cacaagatgg gcgaacagct gcttggagaa
     2701 ctacaagtga agaaaagaaa gcattagatc aggcaagtga agagatttgg aatgattttc
     2761 gagaagctgc agaagcacat cgacaagtta gaaaatacgt aatgagctgg atcaagcctg
     2821 ggatgacaat gatagaaatc tgtgaaaagt tggaagactg ttcacgcaag ttaataaaag
     2881 agaatggatt aaatgcaggc ctggcatttc ctactggatg ttctctcaat aattgtgctg
     2941 cccattatac tcccaatgcc ggtgacacaa cagtattaca gtatgatgac atctgtaaaa
     3001 tagactttgg aacacatata agtggtagga ttattgactg tgcttttact gtcactttta
     3061 atcccaaata tgatacgtta ttaaaagctg taaaagatgc tactaacact ggaataaagt
     3121 gtgctggaat tgatgttcgt ctgtgtgatg ttggtgaggc catccaagaa gttatggagt
     3181 cctatgaagt tgaaatagat gggaagacat atcaagtgaa accaatccgt aatctaaatg
     3241 gacattcaat tgggcaatat agaatacatg ctggaaaaac agtgccgatt gtgaaaggag
     3301 gggaggcaac aagaatggag gaaggagaag tatatgcaat tgaaaccttt ggtagtacag
     3361 gaaaaggtgt tgttcatgat gatatggaat gttcacatta catgaaaaat tttgatgttg
     3421 gacatgtgcc aataaggctt ccaagaacaa aacacttgtt aaatgtcatc aatgaaaact
     3481 ttggaaccct tgccttctgc cgcagatggc tggatcgctt gggagaaagt aaatacttga
     3541 tggctctgaa gaatctgtgt gacttgggca ttgtagatcc atatccacca ttatgtgaca
     3601 ttaaaggatc atatacagcg caatttgaac ataccatcct gttgcgtcca acatgtaaag
     3661 aagttgtcag cagaggagat gactattaag aattctgcag tcgacggtac cgcgggcccg
     3721 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
     3781 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     3841 cgccagcaca gtggtcgacc ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3901 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3961 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4021 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4081 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     4141 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4201 agcttcatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct
     4261 acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc
     4321 gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa
     4381 ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg
     4441 atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc
     4501 gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt
     4561 gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaagatcg ataaaataaa
     4621 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     4681 agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga
     4741 agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg
     4801 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc
     4861 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc
     4921 cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc
     4981 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct
     5041 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg
     5101 gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg
     5161 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
     5221 tacccgtcag cgggggtctt tcacatgcag catgtatcaa aattaatttg gttttttttc
     5281 ttaagtattt acattaaatg gccatagttg cattaatgaa tcggccaacg cgcggggaga
     5341 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5401 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5461 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5521 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5581 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5641 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5701 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5761 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5821 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5881 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5941 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6001 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6061 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6121 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6181 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6241 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6301 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6361 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6421 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6481 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6541 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6601 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6661 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6721 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6781 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6841 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6901 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6961 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7021 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7081 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7141 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7201 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7261 gttccgcgca catttccccg aaaagtgcca cctgcagc
//
