LOCUS       sequence-471181-        8706 bp    DNA     circular SYN 03-JUN-2025
DEFINITION  Express NUP98::LNP1 in bicistronic retroviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8706)
  TITLE     Mullighan lab plasmids 2025
REFERENCE   2  (bases 1 to 8706)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 3, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: CL20_MSCV-NUP98::LNP1-ires-GFP
FEATURES             Location/Qualifiers
     source          1..8706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     LTR             1129..1643
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(join(1745..2458,2459..2461,2462..2464))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1790..1811)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2139..2158
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2398..2419
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    2476..3028
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2478..3045
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2638..2657)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     2867..2884
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     protein_bind    3058..3082
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3662..3679
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(5288..5312)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(5339..5755)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(5355..5371)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(5387..5409)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    5822..6163
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             6227..6743
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(6943..6962)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(7090..7112)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7306..7325)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7425..7447
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7485..7503)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7571..7675
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7676..7744,7745..8536)
                     /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     complement(7894..7913)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacaattctc cccagctctt tctgcgctgt ccataatgaa ctatttcagg
     1021 agtattggag tatgtggatt caaaacagtg gctaattgag tcaggattac ttaatattcg
     1081 actcctaccc tcagaatgtt gttctccaac ttcaagaaac tgttacccat gaaagacccc
     1141 cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga gaccacaagt
     1201 cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgacg cagtctatcg
     1261 gaggactggc gcgccgagtg aggggttgtg ggctctttta ttgagctcgg ggagcagaag
     1321 cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca cagggtcatt
     1381 tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc tgagggcggg
     1441 accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg cttaccacag
     1501 atatcctgtt tggcccatat tctgctgtct ctctgttcct aaccttgatc tgaacttctc
     1561 tattctcagt tatgtatttt ccatgccttg caaaatggcg ttacttaagc tagcttgcca
     1621 aacctacagg tggggtcttt cattcccccc tttttctgga gactaaataa aatcttttat
     1681 tttatcgata agcttggctg caggtcgacg gtatcgataa gcttgataat tcttaattaa
     1741 ttcattactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca
     1801 gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac tgggtgctca
     1861 ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt
     1921 ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcgccttga
     1981 tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca
     2041 gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt
     2101 tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct
     2161 tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt
     2221 gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca
     2281 cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct
     2341 tgccgtaggt ggcatcgccc tcgccctcgc cagacacgct gaacttgtgg ccgtttacgt
     2401 cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca
     2461 ccatggttgt ggccatatta tcatcgtgtt tttcaaagga aaaccacgtc cccgtggttc
     2521 ggggggccta gacgtttttt taacctcgac taaacacatg taaagcatgt gcaccgaggc
     2581 cccagatcag atcccataca atggggtacc ttctgggcat ccttcagccc cttgttgaat
     2641 acgcttgagg agagccattt gactctttcc acaactatcc aactcacaac gtggcactgg
     2701 ggttgtgccg cctttgcagg tgtatcttat acacgtggct tttggccgca gaggcacctg
     2761 tcgccaggtg gggggttccg ctgcctgcaa agggtcgcta cagacgttgt ttgtcttcaa
     2821 gaagcttcca gaggaactgc ttccttcacg acattcaaca gaccttgcat tcctttggcg
     2881 agaggggaaa gacccctagg aatgctcgtc aagaagacag ggccaggttt ccgggccctc
     2941 acattgccaa aagacggcaa tatggtggaa aataacatat agacaaacgc acaccggcct
     3001 tattccaagc ggcttcggcc agtaacgtta gggggggggg gcggaattcg ttatcgaacc
     3061 actttgtaca agaaagctgg gtcggcgcgc ccaccctttt attcaggccc tttttcaaac
     3121 aggggagcca tgtgtgcttc tccatgttct tctttcctag agctcctttc ttcctctact
     3181 ttctttcggg attttatctc catcctcagg cattctcttt catttctctc ctttctgctt
     3241 tcaggtccca aagaggcaga acgcttggtt ctaaagcaca gttgccgttc aaaggactct
     3301 gaaaatttct caattttgga atgggatctt ccttttgatt ccagtggctc cttgaatgac
     3361 ccatcctctg agtattttct gtaatcccgt acgtggctac ggcatcgaaa ttcttggtcc
     3421 tcatgagaat gccttctagg aagacggtca gacgatcggc atcggaattc ctggtcctca
     3481 tgagaatgcc ttctaggatg gcagtcagat gatggaattc tgggaagcac aggaagtggg
     3541 cagggcaggg aggttttcct gtgactggtg gcttgcagtc ggtggcgttc ccggagtcct
     3601 ctctgatcct cctctctcca gctgtggccc cagaagctgc tcatccattt ggcaaaagac
     3661 acatcatcat catcatcatc tttgtgctcc atgtaaaggt gatgccaaat actgaaggcc
     3721 tgtccctctt ccttcttctt agggtctgac atcggattcc ggaagagagg agagtctcca
     3781 aaaggtgagt atgttagact attgatgtgc tgctggagaa cagcctgctg ggcagcagaa
     3841 gcatttggat ctgtcaaagc tactggggcc tggggggctc caaagcccaa agtggctgtc
     3901 gtagtattaa atccaggggc cccaaaggct cctgtaccaa gaggccctcc aatcttaggt
     3961 tggttgttcc caaacaaaga tgcctgtcca gcaccaagag ctgttccaaa tcctgcacca
     4021 agcccagttc caagagtccc aggtgctggt ttacttccaa aaatactatt cccactggta
     4081 tttgtgccaa aaccaaagag cccgccactg gttgtaccaa atgaaggtgc actggttgtg
     4141 ctgcttccaa atgtagtaag cttgttattg ccaaacaggg tcgaaccaac agcaccaaat
     4201 ccagtattgg tctgcccaaa gagacctgtt cctgttccaa atgctgtccc agtgctggtg
     4261 tttgtagctg tcccaaaaag acctccaggc tgtgaggctt gggttactcc aaataatccc
     4321 atggtgttgg tgcttggctg tcctatggtg ctggtattac caaaggaaaa gccagtgttc
     4381 tgggtggttg tagcctggcc aaatggtttg ctgaagaggc tggtagtctg ctgattctgt
     4441 tggccaaaga gaccacctgg atttgttcca aatccagttg tactagttcc aaaggcagtt
     4501 ttgttctgac catatgcaaa gcctgaatta gtggtggagg agctgaagag tcctgttgcg
     4561 ctggaagtgg ctggagaaga cccaaacaag ccagttgtgg tacctgctcc cacctggttc
     4621 tgtgggccct tcctgttagc ctgataatcc tctaaacgaa gttcctctag tgacttgctt
     4681 tcatattctt tcatagcagt aatacactgg tgcttggtac ttatgttagt gctaactcca
     4741 gctttgacca tagtatctgt accagttgga gggttaaatt taatagtagt cccagtagga
     4801 gcagctgtaa aactacttgg cccaaagagg gagccagatg tgctgccaaa aggattagag
     4861 gtggtatttg tggttccaaa gagtcctcca ctgctagtac tggttccaaa attgccaaag
     4921 ccagttggtt tattttgtgc aaaggcattg ttttgggatg agaagagact ggtccctgtg
     4981 cttgcagttc caaacaaggt atttgctgtt cctgttgacg taccaaaccc aaagccagtg
     5041 cttgtggagg tagctggctg gctaaatgaa ctggttccaa acaatcctcc tggtttagtc
     5101 tgtgaatttc caaagaggcc tccagtattg ttgctagaac caaatgcaga tgttccaaat
     5161 gcccctccac tagtagtgcc aaagccagta ttctgtccaa atgttgaagt tgtgccaaag
     5221 ccacctgtgc cacccccaaa gggtgttcca aatgatttgt taaacatggt gaagggggcg
     5281 gccgcggagc ctgctttttt gtacaaactt gtttgataac ctcgagagat ctaattccgg
     5341 cgcctagaga aggagtgagg gctggataaa gggaggatcg aggcggggtc gaacgaggag
     5401 gttcaagggg gagagacggg gcggatggag gaagaggagg cggaggctta gggtgtacaa
     5461 agggcttgac ccagggaggg gggtcaaaag ccaaggcttc ccaggtcacg atgtagggga
     5521 cctggtctgg gtgtccatgc gggccaggtg aaaagacctt gatcttaacc tgggtgatga
     5581 ggtctcggtt aaaggtgccg tctcgcggcc atccgacgtt aaaggttggc cattctgcag
     5641 agcagaaggt aacccaacgt ctcttcttga catctaccga ctggttgtga gcgatccgct
     5701 cgacatcttt ccagtgacct aaggtcaaac ttaagggagt ggtaacagtc tggccctaat
     5761 tttcagacaa atacagaaac acagtcagac agagacaaca cagaacgatg ctgcagcgct
     5821 gcagcagaca agacgcgcgg cttcggttcc aaaccgaaag caaaaattca gacggaggcg
     5881 ggaactgttt taggttctcg tctcctacca gaaccacata tcctgacggg gtcggattcc
     5941 acatcgactc ccttcctcag gtcgggccac aaaaacggcc cccaaagtcc ctgggacgtc
     6001 tcccagggtt gcggccgggt gttcagaact cgtcagttcc accacgggtc cgccagatac
     6061 agagctagtt agctaactag tacagacgca ggcgcaaaca ttagatgccg gcacaaacac
     6121 gcacaaagac agagacagac acgaaacgac cgctggccag cttacctccc ggcggggggg
     6181 tcggtggtcc ctgggcaggg gtctccaaat ctcggtggaa cctccaaatg aaagaccccc
     6241 gaggtgggca gtcaatcaat ctgaggagac cctcccaagg atcagcgagt ccacgattcg
     6301 gatgcaaaca gcaagaggct ttattgggaa tacgggtacc cgggcgacgc agtctatcgg
     6361 aggactggcg cgccgagtga ggggttgtgg gctcttttat tgagctcggg gagcagaagc
     6421 gcgcgaacag aagcgagaag cgaactgatt ggttagttca aataaggcac agggtcattt
     6481 caggtccttg gggcaccctg gaaacatctg atggttctct agaaactgct gagggcggga
     6541 ccgcatctgg ggaccatctg ttcttggccc tgagccgggg caggaactgc ttaccacaga
     6601 tatcctgttt ggcccatatt ctgctgtctc tctgttccta accttgatct gaacttctct
     6661 attctcagtt atgtattttc catgccttgc aaaatggcgt tacttaagct agcttgccaa
     6721 acctacaggt ggggtctttc attcccccct ttttctggag actaaataaa atcttttatt
     6781 ttatctatgg ctcgtactct ataggcttca gctggtgata ttgttgagtc aaaactagag
     6841 cctggaccac tgatatcctg tctttaacaa attggactaa tcgcctctac gccggacgca
     6901 tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca
     6961 ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta
     7021 tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc
     7081 ttgcgccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     7141 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     7201 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     7261 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     7321 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     7381 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     7441 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     7501 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     7561 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     7621 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     7681 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     7741 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     7801 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     7861 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     7921 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     7981 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     8041 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     8101 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     8161 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     8221 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     8281 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     8341 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     8401 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     8461 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     8521 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     8581 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     8641 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     8701 atcttc
//
