LOCUS       sequence-271352-        6874 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 6874)
  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 6874)
  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: pJM010
FEATURES             Location/Qualifiers
     source          1..6874
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(6..110)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             141..610
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    675..874
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1075..1491
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1421..1443
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1459..1475
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1526..1535
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1532..1534,1535..1537,1538..2248)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1532..1534,1535..1537,1538..2248)
                     /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(1577..1598)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1838..1857)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2185..2206
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2294..2314
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2327..2371
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3082..3093
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(3592..3612)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3597..3926
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3777..3912
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3934..4332
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4501..4970
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5100..5117)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5271..5859)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5351..5370)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6019..6810,6811..6874,1..5))
                     /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     6642..6661
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
       61 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
      121 cgaaaagtgc cacctgcagc ctgaatatgg gccaaacagg atatctgtgg taagcagttc
      181 ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc
      241 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
      301 cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
      361 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
      421 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggggcg ccagtcctcc
      481 gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga
      541 cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg
      601 gggtctttca tttgggggct cgtccgggat cgggagaccc ctgcccaggg accaccgacc
      661 caccaccggg aggcaagctg gccagcaact tatctgtgtc tgtccgattg tctagtgtct
      721 atgactgatt ttatgcgcct gcgtcggtac tagttagcta actagctctg tatctggcgg
      781 acccgtggtg gaactgacga gttctgaaca cccggccgca accctgggag acgtcccagg
      841 gactttgggg gccgtttttg tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc
      901 ccgtcaggat atgtggttct ggtaggagac gagaacctaa aacagttccc gcctccgtct
      961 gaatttttgc tttcggtttg gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc
     1021 atcgttctgt gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa ttagggccag
     1081 actgttacca ctcccttaag tttgacctta ggtcactgga aagatgtcga gcggatcgct
     1141 cacaaccagt cggtagatgt caagaagaga cgttgggtta ccttctgctc tgcagaatgg
     1201 ccaaccttta acgtcggatg gccgcgagac ggcaccttta accgagacct catcacccag
     1261 gttaagatca aggtcttttc acctggcccg catggacacc cagaccaggt cccctacatc
     1321 gtgacctggg aagccttggc ttttgacccc cctccctggg tcaagccctt tgtacaccct
     1381 aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg
     1441 accccgcctc gatcctccct ttatccagcc ctcactcctt ctctaggcgc cggccggatc
     1501 ccagtgtggt ggtacgctac cggtcgccac catggtgagc aagggcgagg agctgttcac
     1561 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1621 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     1681 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     1741 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1801 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     1861 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     1921 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     1981 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     2041 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     2101 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     2161 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     2221 cactctcggc atggacgagc tgtacaagtc cggaggggat cggtggagct ccaccggtgg
     2281 cggccgctct agagagaatc tttattttca gggcgccgcc aaattcaaag aaaccgctgc
     2341 tgctaaattc gaacgccagc acatggacag cggaggtgga ggtactagtc tcgagatgga
     2401 ggactccgga aagactttca gctccgagga ggaagaagct aactattgga aagatctggc
     2461 gatgacctac aaacagaggg cagaaaatac gcaagaggaa ctccgagaat tccaggaggg
     2521 aagccgagaa tatgaagctg aattggagac gcagctgcaa caaattgaaa ccaggaacag
     2581 agacctcctg tccgaaaata accgccttcg catggagctg gaaaccatca aggagaagtt
     2641 tgaagtgcag cactctgaag gctaccggca gatctcagcc ttggaggatg acctcgcgca
     2701 gaccaaagcc attaaagacc aattgcagaa atacatcaga gagctggagc aagcaaatga
     2761 cgacctggaa agagccaagc gcgccacgat catgtctctc gaagactttg agcagcgctt
     2821 gaatcaggcc atcgaaagaa atgccttcct ggaaagtgaa cttgatgaaa aagagaatct
     2881 cctggaatct gttcagagac tgaaggatga agccagagat ttgcggcagg aactggccgt
     2941 gcagcagaag caggagaaac ccaggacccc catgcccagc tcagtggaag ctgagaggac
     3001 agacacagct gtgcaggcca cgggctccgt gccgtccacg cccattgctc accgaggacc
     3061 aagctcaagt ttaaacacac ctgggagctt cagacgtggc ctggacgact ccaccggggg
     3121 gacccccctc acacctgcgg cccggatatc agccctcaac attgtgggag acctactgcg
     3181 gaaagtcggg gcactggagt ccaaactcgc ttcctgccgg aacctcgtgt acgatcagtc
     3241 cccaaaccga acaggtggcc cagcctctgg gcggagcagc aagaacagag atggcgggga
     3301 gagacggcca agcagcacca gcgtgccttt gggtgataag gggttgggca agcggctgga
     3361 gttcggcaag ccccctagcc acatgagctc ctctcccctg cccagcgccc agggcgtggt
     3421 caagatgctg ctctgaccgc gggcccggga tccaccggat ctagataact gatcataatc
     3481 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     3541 aacctgaaac ataaaatgaa tgcaattcgc cagcacagtg gtcgaccctg tggaatgtgt
     3601 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3661 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3721 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3781 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3841 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3901 tttttggagg cctaggcttt tgcaaaaagc ttcatggcca agcctttgtc tcaagaagaa
     3961 tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac
     4021 agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat
     4081 cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca
     4141 gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc
     4201 tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag
     4261 gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg
     4321 tgggagggct aagatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     4381 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     4441 ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag
     4501 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     4561 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     4621 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
     4681 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     4741 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     4801 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
     4861 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
     4921 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca catgcagcat
     4981 gtatcaaaat taatttggtt ttttttctta agtatttaca ttaaatggcc atagttgcat
     5041 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5101 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5161 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5221 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5281 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5341 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5401 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5461 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5521 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5581 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5641 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5701 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5761 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5821 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5881 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5941 tcaaaaagga tcttcaccta gatccttttg cggccgcaaa tcaatctaaa gtatatatga
     6001 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6061 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6121 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6181 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6241 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6301 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6361 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6421 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6481 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6541 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6601 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6661 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6721 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6781 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6841 aaagggaata agggcgacac ggaaatgttg aata
//
