LOCUS       sequence-271753-        6861 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 6861)
  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 6861)
  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: pJM147
FEATURES             Location/Qualifiers
     source          1..6861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             54..470
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     400..422
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     438..454
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      505..514
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(511..513,514..516,517..1227)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(511..513,514..516,517..1227)
                     /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(556..577)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(817..836)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1164..1185
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1273..1293
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1306..1350
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2558..2578)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2563..2892
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2743..2878
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2900..3298
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3467..3936
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4066..4083)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4237..4825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4317..4336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4985..5776,5777..5845))
                     /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     5608..5627
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5846..5950)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5981..6450
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6515..6714
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt gtctgaaaat tagggccaga
       61 ctgttaccac tcccttaagt ttgaccttag gtcactggaa agatgtcgag cggatcgctc
      121 acaaccagtc ggtagatgtc aagaagagac gttgggttac cttctgctct gcagaatggc
      181 caacctttaa cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc atcacccagg
      241 ttaagatcaa ggtcttttca cctggcccgc atggacaccc agaccaggtc ccctacatcg
      301 tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt gtacacccta
      361 agcctccgcc tcctcttcct ccatccgccc cgtctctccc ccttgaacct cctcgttcga
      421 ccccgcctcg atcctccctt tatccagccc tcactccttc tctaggcgcc ggccggatcc
      481 cagtgtggtg gtacgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
      541 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
      601 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
      661 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
      721 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
      781 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
      841 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
      901 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
      961 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     1021 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1081 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     1141 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1201 actctcggca tggacgagct gtacaagtcc ggaggggatc ggtggagctc caccggtggc
     1261 ggccgctcta gagagaatct ttattttcag ggcgccgcca aattcaaaga aaccgctgct
     1321 gctaaattcg aacgccagca catggacagc ggaggtggag gtactagtct cgagatggaa
     1381 gatagcggca aaaccttcag cagcgaagaa gaagaggcga attactggaa ggacctggcc
     1441 atgacctaca agcagcgtgc cgaaaatacc caggaagaac tgcgcgaatt tcaagagggg
     1501 agccgcgaat atgccgccga gttggaaaca cagctgcagc agatcgagac acgcaatcgc
     1561 gatctgctga gcgaaaataa ccgcctgcgc atggaactgg aaacaattaa agagaaattc
     1621 gaggtgcagc atagcgaggg ctatcgccaa attagcgccc tggaggatga tctggcccag
     1681 accaaagcca tcaaagatca gctgcagaaa tacattcgcg agctggagca ggcaaatgat
     1741 gatctggaac gcgccaaacg cgccaccatt atgagtctgg aggactttga gcagcgtctg
     1801 aaccaggcaa ttgagcgcaa tgccttcctg gaaagcgaat tagatgagaa agagaatctg
     1861 ttagagagtg tgcagcgcct gaaagacgaa gcccgtgacc tgcgccagga gctggcagtg
     1921 cagcagaagc aagaaaagcc gcgcaccccg atgccgagta gtgttgaagc agaacgtacc
     1981 gataccgcag tgcaagccac cggcagcgtt ccgagtacac cgattgccca tcgcggcccg
     2041 agtagcagtc tgaatacccc gggcagcttt cgtcgtggtc tggatgatag caccggtggc
     2101 acaccgttaa caccggccgc ccgtattagc gccctgaata ttgtgggcga tctgctgcgc
     2161 aaagtgggcg cactggagag taaactggcc agttgccgca acctggtgta tgatcagagc
     2221 ccgaatcgta caggtggtcc ggcaagcggc cgcagtagta aaaatcgtga tggcggcgaa
     2281 cgccgtccga gtagtaccag cgttccgctg ggcgacaaag gcctggatac cagctgccgt
     2341 tggctgagca aaagcaccac ccgcagcagt agcagttgtt aagaattctg cagtcgacgg
     2401 taccgcgggc ccgggatcca ccggatctag ataactgatc ataatcagcc ataccacatt
     2461 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2521 aatgaatgca attcgccagc acagtggtcg accctgtgga atgtgtgtca gttagggtgt
     2581 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2641 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2701 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2761 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2821 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2881 ggcttttgca aaaagcttca tggccaagcc tttgtctcaa gaagaatcca ccctcattga
     2941 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc
     3001 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg
     3061 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac
     3121 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg
     3181 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca
     3241 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaaga
     3301 tcgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc
     3361 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa
     3421 ctgagaatag agaagttcag atcaaggtca ggaacagatg gaacagctga atatgggcca
     3481 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggaaca
     3541 gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     3601 agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg
     3661 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     3721 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     3781 cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca
     3841 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     3901 ctgagtgatt gactacccgt cagcgggggt ctttcacatg cagcatgtat caaaattaat
     3961 ttggtttttt ttcttaagta tttacattaa atggccatag ttgcattaat gaatcggcca
     4021 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4081 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4141 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4201 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4261 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4321 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4381 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4441 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4501 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4561 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4621 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4681 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4741 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4801 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4861 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4921 cacctagatc cttttgcggc cgcaaatcaa tctaaagtat atatgagtaa acttggtctg
     4981 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5041 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5101 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5161 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5221 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5281 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5341 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5401 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5461 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5521 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5581 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5641 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5701 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5761 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5821 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5881 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5941 gggttccgcg cacatttccc cgaaaagtgc cacctgcagc ctgaatatgg gccaaacagg
     6001 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa
     6061 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     6121 gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca
     6181 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
     6241 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
     6301 actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac
     6361 cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt
     6421 gattgactac ccgtcagcgg gggtctttca tttgggggct cgtccgggat cgggagaccc
     6481 ctgcccaggg accaccgacc caccaccggg aggcaagctg gccagcaact tatctgtgtc
     6541 tgtccgattg tctagtgtct atgactgatt ttatgcgcct gcgtcggtac tagttagcta
     6601 actagctctg tatctggcgg acccgtggtg gaactgacga gttctgaaca cccggccgca
     6661 accctgggag acgtcccagg gactttgggg gccgtttttg tggcccgacc tgaggaaggg
     6721 agtcgatgtg gaatccgacc ccgtcaggat atgtggttct ggtaggagac gagaacctaa
     6781 aacagttccc gcctccgtct gaatttttgc tttcggtttg gaaccgaagc cgcgcgtctt
     6841 gtctgctgca gcgctgcagc a
//
