LOCUS       sequence-475549-        8274 bp    DNA     circular SYN 10-JUN-2025
DEFINITION  Express NUP98::HOXA9 in bicistronic retroviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8274)
  TITLE     Mullighan lab plasmids 2025
REFERENCE   2  (bases 1 to 8274)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 10, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: CL20_MSCV-NUP98::HOXA9-ires-GFP
FEATURES             Location/Qualifiers
     source          1..8274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(98..117)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      911..1499
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1400..1419
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1653..1670
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1787..1808
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1823..1828,1829..1846,1847..1853)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1861..1877
                     /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             2039..2553
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(join(2655..3368,3369..3371,3372..3374))
                     /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(2700..2721)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3049..3068
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3308..3329
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    3386..3938
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3388..3955
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3548..3567)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     3777..3794
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     protein_bind    3968..3992
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5766..5790)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(5817..6233)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(5833..5849)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(5865..5887)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    6300..6641
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             6705..7221
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(7421..7440)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(7568..7590)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7784..7803)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7903..7925
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7963..7981)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8049..8153
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8154..8222,8223..8274,1..740)
                     /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"
ORIGIN
        1 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
       61 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
      121 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
      181 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
      241 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
      301 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
      361 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
      421 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
      481 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
      541 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
      601 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
      661 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
      721 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
      781 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
      841 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
      901 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      961 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1021 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     1081 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1141 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1201 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1261 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1321 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1381 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     1441 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     1501 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     1561 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     1621 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     1681 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     1741 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     1801 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     1861 ttgtgagcgg ataacaattt cacaattctc cccagctctt tctgcgctgt ccataatgaa
     1921 ctatttcagg agtattggag tatgtggatt caaaacagtg gctaattgag tcaggattac
     1981 ttaatattcg actcctaccc tcagaatgtt gttctccaac ttcaagaaac tgttacccat
     2041 gaaagacccc cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga
     2101 gaccacaagt cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgacg
     2161 cagtctatcg gaggactggc gcgccgagtg aggggttgtg ggctctttta ttgagctcgg
     2221 ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca
     2281 cagggtcatt tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc
     2341 tgagggcggg accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg
     2401 cttaccacag atatcctgtt tggcccatat tctgctgtct ctctgttcct aaccttgatc
     2461 tgaacttctc tattctcagt tatgtatttt ccatgccttg caaaatggcg ttacttaagc
     2521 tagcttgcca aacctacagg tggggtcttt cattcccccc tttttctgga gactaaataa
     2581 aatcttttat tttatcgata agcttggctg caggtcgacg gtatcgataa gcttgataat
     2641 tcttaattaa ttcattactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc
     2701 acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac
     2761 tgggtgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc
     2821 tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag
     2881 ttcgccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag
     2941 ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc
     3001 tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg
     3061 ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg
     3121 aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc
     3181 agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc
     3241 agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cagacacgct gaacttgtgg
     3301 ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg
     3361 cccttgctca ccatggttgt ggccatatta tcatcgtgtt tttcaaagga aaaccacgtc
     3421 cccgtggttc ggggggccta gacgtttttt taacctcgac taaacacatg taaagcatgt
     3481 gcaccgaggc cccagatcag atcccataca atggggtacc ttctgggcat ccttcagccc
     3541 cttgttgaat acgcttgagg agagccattt gactctttcc acaactatcc aactcacaac
     3601 gtggcactgg ggttgtgccg cctttgcagg tgtatcttat acacgtggct tttggccgca
     3661 gaggcacctg tcgccaggtg gggggttccg ctgcctgcaa agggtcgcta cagacgttgt
     3721 ttgtcttcaa gaagcttcca gaggaactgc ttccttcacg acattcaaca gaccttgcat
     3781 tcctttggcg agaggggaaa gacccctagg aatgctcgtc aagaagacag ggccaggttt
     3841 ccgggccctc acattgccaa aagacggcaa tatggtggaa aataacatat agacaaacgc
     3901 acaccggcct tattccaagc ggcttcggcc agtaacgtta gggggggggg gcggaattcg
     3961 ttatcgaacc actttgtaca agaaagctgg gtcggcgcgc ccaccctttc actcgtcttt
     4021 tgctcggtct ttgttgattt tcttcatttt catcctgcgg ttctggaacc agatcttgac
     4081 ctgcctctcg gtgaggttga gcagtcgagc cacctcgtac ctgcggtccc tggtgaggta
     4141 catgttgaac agaaactctt tctccagttc cagggtctgg tgttttgtat aggggcaccg
     4201 ctttttccga gtggagcgcg catgaagcca gttggctgct gggttattgg gatcgatggg
     4261 gggcttgtct ccgccgctct cattctcagc attgttttca gagaaggcgc cttcgctggg
     4321 ttgtttttct ctatcaacta ctggggcctg gggggctcca aagcccaaag tggctgtcgt
     4381 agtattaaat ccaggggccc caaaggctcc tgtaccaaga ggccctccaa tcttaggttg
     4441 gttgttccca aacaaagatg cctgtccagc accaagagct gttccaaatc ctgcaccaag
     4501 cccagttcca agagtcccag gtgctggttt acttccaaaa atactattcc cactggtatt
     4561 tgtgccaaaa ccaaagagcc cgccactggt tgtaccaaat gaaggtgcac tggttgtgct
     4621 gcttccaaat gtagtaagct tgttattgcc aaacagggtc gaaccaacag caccaaatcc
     4681 agtattggtc tgcccaaaga gacctgttcc tgttccaaat gctgtcccag tgctggtgtt
     4741 tgtagctgtc ccaaaaagac ctccaggctg tgaggcttgg gttactccaa ataatcccat
     4801 ggtgttggtg cttggctgtc ctatggtgct ggtattacca aaggaaaagc cagtgttctg
     4861 ggtggttgta gcctggccaa atggtttgct gaagaggctg gtagtctgct gattctgttg
     4921 gccaaagaga ccacctggat ttgttccaaa tccagttgta ctagttccaa aggcagtttt
     4981 gttctgacca tatgcaaagc ctgaattagt ggtggaggag ctgaagagtc ctgttgcgct
     5041 ggaagtggct ggagaagacc caaacaagcc agttgtggta cctgctccca cctggttctg
     5101 tgggcccttc ctgttagcct gataatcctc taaacgaagt tcctctagtg acttgctttc
     5161 atattctttc atagcagtaa tacactggtg cttggtactt atgttagtgc taactccagc
     5221 tttgaccata gtatctgtac cagttggagg gttaaattta atagtagtcc cagtaggagc
     5281 agctgtaaaa ctacttggcc caaagaggga gccagatgtg ctgccaaaag gattagaggt
     5341 ggtatttgtg gttccaaaga gtcctccact gctagtactg gttccaaaat tgccaaagcc
     5401 agttggttta ttttgtgcaa aggcattgtt ttgggatgag aagagactgg tccctgtgct
     5461 tgcagttcca aacaaggtat ttgctgttcc tgttgacgta ccaaacccaa agccggtgct
     5521 tgtggaggta gctggctggc taaatgaact ggttccaaac aatcctcctg gtttagtctg
     5581 tgaatttcca aagaggcctc cagtattgtt gctagaacca aatgcagatg ttccaaatgc
     5641 ccctccacta gtagtgccaa agccagtatt ctgtccaaat gttgaagttg tgccaaagcc
     5701 acctgtgcca cccccaaagg gtgttccaaa tgatttgtta aacatggtga agggggcggc
     5761 cgcggagcct gcttttttgt acaaacttgt ttgataacct cgagagatct aattccggcg
     5821 cctagagaag gagtgagggc tggataaagg gaggatcgag gcggggtcga acgaggaggt
     5881 tcaaggggga gagacggggc ggatggagga agaggaggcg gaggcttagg gtgtacaaag
     5941 ggcttgaccc agggaggggg gtcaaaagcc aaggcttccc aggtcacgat gtaggggacc
     6001 tggtctgggt gtccatgcgg gccaggtgaa aagaccttga tcttaacctg ggtgatgagg
     6061 tctcggttaa aggtgccgtc tcgcggccat ccgacgttaa aggttggcca ttctgcagag
     6121 cagaaggtaa cccaacgtct cttcttgaca tctaccgact ggttgtgagc gatccgctcg
     6181 acatctttcc agtgacctaa ggtcaaactt aagggagtgg taacagtctg gccctaattt
     6241 tcagacaaat acagaaacac agtcagacag agacaacaca gaacgatgct gcagcgctgc
     6301 agcagacaag acgcgcggct tcggttccaa accgaaagca aaaattcaga cggaggcggg
     6361 aactgtttta ggttctcgtc tcctaccaga accacatatc ctgacggggt cggattccac
     6421 atcgactccc ttcctcaggt cgggccacaa aaacggcccc caaagtccct gggacgtctc
     6481 ccagggttgc ggccgggtgt tcagaactcg tcagttccac cacgggtccg ccagatacag
     6541 agctagttag ctaactagta cagacgcagg cgcaaacatt agatgccggc acaaacacgc
     6601 acaaagacag agacagacac gaaacgaccg ctggccagct tacctcccgg cgggggggtc
     6661 ggtggtccct gggcaggggt ctccaaatct cggtggaacc tccaaatgaa agacccccga
     6721 ggtgggcagt caatcaatct gaggagaccc tcccaaggat cagcgagtcc acgattcgga
     6781 tgcaaacagc aagaggcttt attgggaata cgggtacccg ggcgacgcag tctatcggag
     6841 gactggcgcg ccgagtgagg ggttgtgggc tcttttattg agctcgggga gcagaagcgc
     6901 gcgaacagaa gcgagaagcg aactgattgg ttagttcaaa taaggcacag ggtcatttca
     6961 ggtccttggg gcaccctgga aacatctgat ggttctctag aaactgctga gggcgggacc
     7021 gcatctgggg accatctgtt cttggccctg agccggggca ggaactgctt accacagata
     7081 tcctgtttgg cccatattct gctgtctctc tgttcctaac cttgatctga acttctctat
     7141 tctcagttat gtattttcca tgccttgcaa aatggcgtta cttaagctag cttgccaaac
     7201 ctacaggtgg ggtctttcat tccccccttt ttctggagac taaataaaat cttttatttt
     7261 atctatggct cgtactctat aggcttcagc tggtgatatt gttgagtcaa aactagagcc
     7321 tggaccactg atatcctgtc tttaacaaat tggactaatc gcctctacgc cggacgcatc
     7381 gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc
     7441 gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg
     7501 gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt
     7561 gcgccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     7621 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     7681 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta
     7741 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     7801 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     7861 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     7921 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     7981 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     8041 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     8101 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     8161 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     8221 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcac
//
