LOCUS       sequence-271471-        6796 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 6796)
  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 6796)
  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: pJM132
FEATURES             Location/Qualifiers
     source          1..6796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             48..517
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    582..781
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             982..1398
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1328..1350
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1366..1382
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1433..1442
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1439..1441,1442..1444,1445..2155)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1439..1441,1442..1444,1445..2155)
                     /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(1484..1505)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1745..1764)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2092..2113
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2201..2221
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2234..2278
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             2989..3000
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(3421..3441)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3426..3755
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3606..3741
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3763..4161
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4330..4799
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4929..4946)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5100..5688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5180..5199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5848..6639,6640..6708))
                     /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     6471..6490
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6709..6796,1..17))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcagcctg aatatgggcc
       61 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
      121 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
      181 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
      241 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
      301 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
      361 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
      421 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
      481 tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg
      541 gagacccctg cccagggacc accgacccac caccgggagg caagctggcc agcaacttat
      601 ctgtgtctgt ccgattgtct agtgtctatg actgatttta tgcgcctgcg tcggtactag
      661 ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc
      721 ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga
      781 ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag
      841 aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc
      901 gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt
      961 ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt
     1021 cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt
     1081 tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc
     1141 acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat
     1201 ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct
     1261 ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg
     1321 tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc
     1381 actccttctc taggcgccgg ccggatccca gtgtggtggt acgctaccgg tcgccaccat
     1441 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1501 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1561 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1621 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     1681 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1741 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1801 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1861 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     1921 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1981 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2041 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2101 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg
     2161 aggggatcgg tggagctcca ccggtggcgg ccgctctaga gagaatcttt attttcaggg
     2221 cgccgccaaa ttcaaagaaa ccgctgctgc taaattcgaa cgccagcaca tggacagcgg
     2281 aggtggaggt actagtctcg agatggagga ctccggaaag actttcagct ccgaggagga
     2341 agaagctaac tattggaaag atctggcgat gacctacaaa cagagggcag aaaatacgca
     2401 agaggaactc cgagaattcc aggagggaag ccgagaatat gaagctgaat tggagacgca
     2461 gctgcaacaa attgaaacca ggaacagaga cctcctgtcc gaaaataacc gccttcgcat
     2521 ggagctggaa accatcaagg agaagtttga agtgcagcac tctgaaggct accggcagat
     2581 ctcagccttg gaggatgacc tcgcgcagac caaagccatt aaagaccaat tgcagaaata
     2641 catcagagag ctggagcaag caaatgacga cctggaaaga gccaagcgcg ccacgatcat
     2701 gtctctcgaa gactttgagc agcgcttgaa tcaggccatc gaaagaaatg ccttcctgga
     2761 aagtgaactt gatgaaaaag agaatctcct ggaatctgtt cagagactga aggatgaagc
     2821 cagagatttg cggcaggaac tggccgtgca gcagaagcag gagaaaccca ggacccccat
     2881 gcccagctca gtggaagctg agaggacaga cacagctgtg caggccacgg gctccgtgcc
     2941 gtccacgccc attgctcacc gaggaccaag ctcaagttta aacacacctg ggagcttcag
     3001 acgtggcctg gacgactcca ccggggggac ccccctcaca cctgcggccc ggatatcagc
     3061 cctcaacatt gtgggagacc tactgcggaa agtcggggca ctggagtcca aactcgcttc
     3121 ctgccggaac ctcgtgtacg atcagtcccc aaaccgaaca ggtggcccag cctctgggcg
     3181 gagcagcaag aacagagatg gcggggagag acggccaagc agcaccagcg tgcctttggg
     3241 tgataagggg ttgggcaagc ggtgaccgcg ggcccgggat ccaccggatc tagataactg
     3301 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3361 ctccccctga acctgaaaca taaaatgaat gcaattcgcc agcacagtgg tcgaccctgt
     3421 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3481 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3541 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3601 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3661 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3721 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tcatggccaa gcctttgtct
     3781 caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct
     3841 gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc
     3901 aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct
     3961 gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc
     4021 ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc
     4081 atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct
     4141 ggttatgtgt gggagggcta agatcgataa aataaaagat tttatttagt ctccagaaaa
     4201 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt
     4261 gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag
     4321 atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     4381 cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca
     4441 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc
     4501 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     4561 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     4621 gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag
     4681 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
     4741 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac
     4801 atgcagcatg tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca
     4861 tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4921 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4981 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5041 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5101 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5161 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5221 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5281 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5341 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5401 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5461 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5521 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5581 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5641 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5701 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5761 atgagattat caaaaaggat cttcacctag atccttttgc ggccgcaaat caatctaaag
     5821 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5881 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5941 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6001 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6061 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6121 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6181 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6241 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6301 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6361 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6421 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6481 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6541 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6601 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6661 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6721 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6781 tatttagaaa aataaa
//
