LOCUS       sequence-271356-        6631 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 6631)
  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 6631)
  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: pJM008
FEATURES             Location/Qualifiers
     source          1..6631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    283..482
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             683..1099
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1029..1051
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1067..1083
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1134..1143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1140..1142,1143..1145,1146..1856)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1140..1142,1143..1145,1146..1856)
                     /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(1185..1206)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1446..1465)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1793..1814
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1902..1922
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1935..1979
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             2690..2701
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(2957..2977)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2962..3291
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3142..3277
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3299..3697
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3866..4335
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4465..4482)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4636..5224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4716..4735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5384..6175,6176..6244))
                     /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     6007..6026
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6245..6349)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(6380..6631,1..218)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
       61 aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
      121 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
      181 ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg
      241 ggagacccct gcccagggac caccgaccca ccaccgggag gcaagctggc cagcaactta
      301 tctgtgtctg tccgattgtc tagtgtctat gactgatttt atgcgcctgc gtcggtacta
      361 gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc
      421 cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg
      481 aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga
      541 gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga accgaagccg
      601 cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
      661 tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg
      721 tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg
      781 ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg
      841 cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca
      901 tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc
      961 tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc
     1021 gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct
     1081 cactccttct ctaggcgccg gccggatccc agtgtggtgg tacgctaccg gtcgccacca
     1141 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1201 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     1261 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     1321 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     1381 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     1441 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     1501 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     1561 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     1621 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     1681 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     1741 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     1801 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtccg
     1861 gaggggatcg gtggagctcc accggtggcg gccgctctag agagaatctt tattttcagg
     1921 gcgccgccaa attcaaagaa accgctgctg ctaaattcga acgccagcac atggacagcg
     1981 gaggtggagg tactagtctc gagatggagg actccggaaa gactttcagc tccgaggagg
     2041 aagaagctaa ctattggaaa gatctggcga tgacctacaa acagagggca gaaaatacgc
     2101 aagaggaact ccgagaattc caggagggaa gccgagaata tgaagctgaa ttggagacgc
     2161 agctgcaaca aattgaaacc aggaacagag acctcctgtc cgaaaataac cgccttcgca
     2221 tggagctgga aaccatcaag gagaagtttg aagtgcagca ctctgaaggc taccggcaga
     2281 tctcagcctt ggaggatgac ctcgcgcaga ccaaagccat taaagaccaa ttgcagaaat
     2341 acatcagaga gctggagcaa gcaaatgacg acctggaaag agccaagcgc gccacgatca
     2401 tgtctctcga agactttgag cagcgcttga atcaggccat cgaaagaaat gccttcctgg
     2461 aaagtgaact tgatgaaaaa gagaatctcc tggaatctgt tcagagactg aaggatgaag
     2521 ccagagattt gcggcaggaa ctggccgtgc agcagaagca ggagaaaccc aggaccccca
     2581 tgcccagctc agtggaagct gagaggacag acacagctgt gcaggccacg ggctccgtgc
     2641 cgtccacgcc cattgctcac cgaggaccaa gctcaagttt aaacacacct gggagcttca
     2701 gacgtggcct ggacgactcc accgggggga cccccctcac acctgcggcc cggatatcag
     2761 ccctcaacat tgtgggagac ctactgcgga aagtcgggtg accgcgggcc cgggatccac
     2821 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2881 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttcgccagca
     2941 cagtggtcga ccctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3001 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3061 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3121 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3181 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3241 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttcat
     3301 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
     3361 cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg
     3421 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
     3481 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
     3541 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
     3601 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
     3661 tgaattgctg ccctctggtt atgtgtggga gggctaagat cgataaaata aaagatttta
     3721 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
     3781 taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga gaagttcaga
     3841 tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc
     3901 agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg gccaaacagg
     3961 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg
     4021 cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac
     4081 ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg
     4141 cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gggcgccagt
     4201 cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca
     4261 tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc
     4321 agcgggggtc tttcacatgc agcatgtatc aaaattaatt tggttttttt tcttaagtat
     4381 ttacattaaa tggccatagt tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4441 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4501 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4561 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4621 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4681 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4741 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4801 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4861 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4921 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4981 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5041 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5101 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5161 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5221 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5281 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttgcggcc
     5341 gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5401 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5461 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5521 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5581 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5641 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5701 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5761 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5821 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5881 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5941 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6001 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6061 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6121 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6181 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6241 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6301 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6361 gaaaagtgcc acctgcagcc tgaatatggg ccaaacagga tatctgtggt aagcagttcc
     6421 tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct
     6481 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     6541 agccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     6601 tgaccctgtg ccttatttga actaaccaat c
//
