LOCUS       sequence-271475-        6516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral plasmid expressing GFP - Nde1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6516)
  AUTHORS   Monda JK, Cheeseman IM
  TITLE     Nde1 promotes diverse dynein functions through differential 
            interactions and exhibits an isoform-specific proteasome 
            association.
  JOURNAL   Mol Biol Cell. 2018 Sep 15;29(19):2336-2345. doi: 
            10.1091/mbc.E18-07-0418. Epub 2018 Jul 19.
  PUBMED    30024347
REFERENCE   2  (bases 1 to 6516)
  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"; doi: 
            "10.1091/mbc.E18-07-0418"; journalName: "Mol Biol Cell"; date: 
            "2018-09-15- 15"; volume: "29"; issue: "19"; pages: "2336-2345"
COMMENT     Sequence Label: pJM143
FEATURES             Location/Qualifiers
     source          1..6516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             68..484
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     414..436
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     452..468
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      519..528
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(525..527,528..530,531..1241)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(525..527,528..530,531..1241)
                     /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(570..591)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(831..850)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1178..1199
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1287..1307
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1320..1364
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2227..2247)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2232..2561
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2412..2547
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2569..2967
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3136..3605
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3735..3752)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3906..4494)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3986..4005)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4654..5445,5446..5514))
                     /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     5277..5296
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5515..5619)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5650..6119
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6184..6383
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga
       61 aaattagggc cagactgtta ccactccctt aagtttgacc ttaggtcact ggaaagatgt
      121 cgagcggatc gctcacaacc agtcggtaga tgtcaagaag agacgttggg ttaccttctg
      181 ctctgcagaa tggccaacct ttaacgtcgg atggccgcga gacggcacct ttaaccgaga
      241 cctcatcacc caggttaaga tcaaggtctt ttcacctggc ccgcatggac acccagacca
      301 ggtcccctac atcgtgacct gggaagcctt ggcttttgac ccccctccct gggtcaagcc
      361 ctttgtacac cctaagcctc cgcctcctct tcctccatcc gccccgtctc tcccccttga
      421 acctcctcgt tcgaccccgc ctcgatcctc cctttatcca gccctcactc cttctctagg
      481 cgccggccgg atcccagtgt ggtggtacgc taccggtcgc caccatggtg agcaagggcg
      541 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      601 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      661 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
      721 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
      781 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
      841 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
      901 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
      961 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     1021 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     1081 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     1141 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1201 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtccggaggg gatcggtgga
     1261 gctccaccgg tggcggccgc tctagagaga atctttattt tcagggcgcc gccaaattca
     1321 aagaaaccgc tgctgctaaa ttcgaacgcc agcacatgga cagcggaggt ggaggtacta
     1381 gtctcgagat ggaagatagc ggcaaaacct tcagcagcga agaagaagag gcgaattact
     1441 ggaaggacct ggccatgacc tacaagcagc gtgccgaaaa tacccaggaa gaactgcgcg
     1501 aatttcaaga ggggagccgc gaatatgagg ccgagctgga aacacagctg cagcagatcg
     1561 agacacgcaa tcgcgatctg ctgagcgaaa ataaccgcct gcgcatggaa ctggaaacaa
     1621 ttaaagagaa attcgaggtg cagcatagcg agggctatcg ccaaattagc gccctggagg
     1681 atgatctggc ccagaccaaa gccatcaaag atcagctgca gaaatacatt cgcgagctgg
     1741 agcaggcaaa tgatgatctg gaacgcgcca aacgcgccac cattatgagt ctggaggact
     1801 ttgagcagcg tctgaaccag gcaattgagc gcaatgcctt cctggaaagc gaattagatg
     1861 agaaagagaa tctgttagag agtgtgcagc gcctgaaaga cgaagcccgt gacctgcgcc
     1921 aggagctggc agtgcagcag aagcaagaaa agccgcgcac cccgatgccg agtagtgttg
     1981 aagcagaacg taccgatacc gcagtgcaag ccaccggcag cgttccgagt acaccgattg
     2041 cccatcgctg agaattctgc agtcgacggt accgcgggcc cgggatccac cggatctaga
     2101 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2161 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttcgccagca cagtggtcga
     2221 ccctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2281 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2341 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     2401 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2461 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     2521 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttcat ggccaagcct
     2581 ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc
     2641 atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact
     2701 ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact
     2761 gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg
     2821 ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc
     2881 aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg
     2941 ccctctggtt atgtgtggga gggctaagat cgataaaata aaagatttta tttagtctcc
     3001 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc
     3061 cattttgcaa ggcatggaaa aatacataac tgagaataga gaagttcaga tcaaggtcag
     3121 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     3181 ccggctcagg gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg
     3241 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc
     3301 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     3361 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     3421 ctccccgagc tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg
     3481 actgagtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt
     3541 ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc
     3601 tttcacatgc agcatgtatc aaaattaatt tggttttttt tcttaagtat ttacattaaa
     3661 tggccatagt tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3721 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3781 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3841 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3901 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3961 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4021 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4081 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4141 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4201 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4261 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4321 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4381 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4441 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4501 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4561 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttgcggcc gcaaatcaat
     4621 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4681 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4741 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4801 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4861 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4921 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4981 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5041 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5101 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5161 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5221 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5281 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5341 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5401 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5461 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5521 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5581 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5641 acctgcagcc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     5701 cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca
     5761 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc
     5821 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     5881 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     5941 gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag
     6001 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
     6061 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat
     6121 ttgggggctc gtccgggatc gggagacccc tgcccaggga ccaccgaccc accaccggga
     6181 ggcaagctgg ccagcaactt atctgtgtct gtccgattgt ctagtgtcta tgactgattt
     6241 tatgcgcctg cgtcggtact agttagctaa ctagctctgt atctggcgga cccgtggtgg
     6301 aactgacgag ttctgaacac ccggccgcaa ccctgggaga cgtcccaggg actttggggg
     6361 ccgtttttgt ggcccgacct gaggaaggga gtcgatgtgg aatccgaccc cgtcaggata
     6421 tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg aatttttgct
     6481 ttcggtttgg aaccgaagcc gcgcgtcttg tctgct
//
