LOCUS       sequence-471182-        8331 bp    DNA     circular SYN 03-JUN-2025
DEFINITION  Express NUP98::PRRX1 in bicistronic retroviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8331)
  TITLE     Mullighan lab plasmids 2025
REFERENCE   2  (bases 1 to 8331)
  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::PRRX1-ires-GFP
FEATURES             Location/Qualifiers
     source          1..8331
                     /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"
     protein_bind    complement(4913..4937)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(4964..5380)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(4980..4996)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(5012..5034)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    5447..5788
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             5852..6368
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(6568..6587)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(6715..6737)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6931..6950)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7050..7072
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7110..7128)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7196..7300
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7301..7369,7370..8161)
                     /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(7519..7538)
                     /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 agaatccgtt atgaagcccc
     3121 tcgtgtaaac aacatcttgg gagggacgag gatctgtacg gagacgctgt cccccaggag
     3181 agataatcgg tgggtctcgg agcaggacga ggtacgatgg gctgctccac agcagtcacg
     3241 tctcctgagt aggatttgag gagggaagcg tttttattgg ctagcatggc tctctcattc
     3301 ctgcggaact tggctcttcg gttctgaaac cacacctgca ctctcgcctc ggtgaggttc
     3361 acccggcggg caaggtcttc tcgcacaaaa gcatcaggat agtgtgtccg ctcaaagaca
     3421 cgctccaaag cctgcagctg gctgctattg aaggttgtcc tattccttcg ctgctttctc
     3481 ttcttttttt cttctgagtt cagctggtca tctactgggg cctggggggc tccaaagccc
     3541 aaagtggctg tcgtagtatt aaatccaggg gccccaaagg ctcctgtacc aagaggccct
     3601 ccaatcttag gttggttgtt cccagacaaa gatgcctgtc cagcaccaag agctgttcca
     3661 aatcctgcac caagcccagt tccaagagtc ccaggtgctg gtttacttcc aaaaatacta
     3721 ttcccactgg tatttgtgcc aaaaccaaag agcccgccat tggttgtacc aaatgaaggt
     3781 gcactggttg tgctgcttcc aaatgtagta agcttgttat tgccaaacag ggtcgaacca
     3841 acagcaccaa atccagtatt ggtctgccca aagagacctg ttcctgttcc aaatgctgtc
     3901 ccagtgctgg tgtttgtagc tgtcccaaaa agacctccag gctgtgaggc ttgggttact
     3961 ccaaataatc ccatggtgtt ggtgcttggc tgtcctatgg tgctggtatt accaaaggaa
     4021 aagccagtgt tctgggtggt tgtagcctgg ccaaatggtt tgctgaagag gctggtagtc
     4081 tgctgattct gttggccaaa gagaccacct ggatttgttc caaatccagt tgtaccatat
     4141 gcaaagcctg aattagtggt ggaggagctg aagagtcctg ttgcgctgga agtggctgga
     4201 gaagacccaa acaagccagt tgtggtacct gctcccacct ggttctgtgg gcccttcctg
     4261 ttagcctgat aatcctctaa acgaagttcc tctagtgact tgctttcata ttctttcata
     4321 gcagtaatac actggtgctt ggtacttatg ttagtgctaa ctccagcttt gaccatagta
     4381 tctgtaccag ttggagggtt aaatttaata gtagtcccag taggagcagc tgtaaaacta
     4441 cttggcccaa agagggagcc agatgtgctg ccaaaaggat tagaggtggt atttgtggtt
     4501 ccaaagagtc ctccactgct agtactggtt ccaaaattgc caaagccagt tggtttattt
     4561 tgtgcaaagg cattgttttg ggatgagaag agactggtcc ctgtgcttgc agttccaaac
     4621 aaggtatttg ctgttcctgt tgacgtacca aacccaaagc cagtgcttgt ggaggtagct
     4681 ggctggctaa atgaactggt tccaaacaat cctcctggtt tagtctgtga atttccaaag
     4741 aggcctccag tattgttgct agaaccaaat gcagatgttc caaatgcccc tccactagta
     4801 gtgccaaagc cagtattctg tccaaatgtt gaagttgtgc caaagccacc tgtgccaccc
     4861 ccaaagggtg ttccaaatga tttgttaaac atggtgaagg gggcggccgc ggagcctgct
     4921 tttttgtaca aacttgtttg ataacctcga gagatctaat tccggcgcct agagaaggag
     4981 tgagggctgg ataaagggag gatcgaggcg gggtcgaacg aggaggttca agggggagag
     5041 acggggcgga tggaggaaga ggaggcggag gcttagggtg tacaaagggc ttgacccagg
     5101 gaggggggtc aaaagccaag gcttcccagg tcacgatgta ggggacctgg tctgggtgtc
     5161 catgcgggcc aggtgaaaag accttgatct taacctgggt gatgaggtct cggttaaagg
     5221 tgccgtctcg cggccatccg acgttaaagg ttggccattc tgcagagcag aaggtaaccc
     5281 aacgtctctt cttgacatct accgactggt tgtgagcgat ccgctcgaca tctttccagt
     5341 gacctaaggt caaacttaag ggagtggtaa cagtctggcc ctaattttca gacaaataca
     5401 gaaacacagt cagacagaga caacacagaa cgatgctgca gcgctgcagc agacaagacg
     5461 cgcggcttcg gttccaaacc gaaagcaaaa attcagacgg aggcgggaac tgttttaggt
     5521 tctcgtctcc taccagaacc acatatcctg acggggtcgg attccacatc gactcccttc
     5581 ctcaggtcgg gccacaaaaa cggcccccaa agtccctggg acgtctccca gggttgcggc
     5641 cgggtgttca gaactcgtca gttccaccac gggtccgcca gatacagagc tagttagcta
     5701 actagtacag acgcaggcgc aaacattaga tgccggcaca aacacgcaca aagacagaga
     5761 cagacacgaa acgaccgctg gccagcttac ctcccggcgg gggggtcggt ggtccctggg
     5821 caggggtctc caaatctcgg tggaacctcc aaatgaaaga cccccgaggt gggcagtcaa
     5881 tcaatctgag gagaccctcc caaggatcag cgagtccacg attcggatgc aaacagcaag
     5941 aggctttatt gggaatacgg gtacccgggc gacgcagtct atcggaggac tggcgcgccg
     6001 agtgaggggt tgtgggctct tttattgagc tcggggagca gaagcgcgcg aacagaagcg
     6061 agaagcgaac tgattggtta gttcaaataa ggcacagggt catttcaggt ccttggggca
     6121 ccctggaaac atctgatggt tctctagaaa ctgctgaggg cgggaccgca tctggggacc
     6181 atctgttctt ggccctgagc cggggcagga actgcttacc acagatatcc tgtttggccc
     6241 atattctgct gtctctctgt tcctaacctt gatctgaact tctctattct cagttatgta
     6301 ttttccatgc cttgcaaaat ggcgttactt aagctagctt gccaaaccta caggtggggt
     6361 ctttcattcc cccctttttc tggagactaa ataaaatctt ttattttatc tatggctcgt
     6421 actctatagg cttcagctgg tgatattgtt gagtcaaaac tagagcctgg accactgata
     6481 tcctgtcttt aacaaattgg actaatcgcc tctacgccgg acgcatcgtg gccggcatca
     6541 ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc
     6601 gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg
     6661 tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg ccgtcgtttt
     6721 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     6781 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     6841 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     6901 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     6961 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     7021 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     7081 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     7141 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     7201 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     7261 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     7321 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     7381 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     7441 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     7501 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     7561 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     7621 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     7681 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     7741 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     7801 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     7861 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     7921 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     7981 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     8041 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     8101 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     8161 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     8221 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     8281 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt c
//
