LOCUS       sequence-181552-        7445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of human HoxB4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7445)
  AUTHORS   Vo BT, Li C, Morgan MA, Theurillat I, Finkelstein D, Wright S, Hyle 
            J, Smith SM, Fan Y, Wang YD, Wu G, Orr BA, Northcott PA, Shilatifard
            A, Sherr CJ, Roussel MF
  TITLE     Inactivation of Ezh2 Upregulates Gfi1 and Drives Aggressive 
            Myc-Driven Group 3 Medulloblastoma.
  JOURNAL   Cell Rep. 2017 Mar 21;18(12):2907-2917. doi: 
            10.1016/j.celrep.2017.02.073.
  PUBMED    28329683
REFERENCE   2  (bases 1 to 7445)
  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: 
            "10.1016/j.celrep.2017.02"; journalName: "Cell Rep"; date: 
            "2017-03-21- 21"; volume: "18"; issue: "12"; pages: "2907-2917"
COMMENT     Sequence Label: MSCV-human HoxB4-IRES-YFP
FEATURES             Location/Qualifiers
     source          1..7445
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    758..1307
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(902..919)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1129..1148
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1310..1312,1313..1315,1316..2029)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1355..1376)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1616..1635)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1963..1984
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2191..2705
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    2867..2883
                     /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)."
     promoter        complement(join(2891..2897,2898..2915,2916..2921))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2936..2957
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3074..3091)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3245..3833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3325..3344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4004..4795,4796..4864))
                     /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     4627..4646
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4865..4969)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5037..5055
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5093..5115)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5215..5234
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5428..5450
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5578..5597
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             5797..6313
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6377..6718
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             6785..7201
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7131..7153
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7169..7185
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 ggcctgccgg gaccctgggc ccccgccgcc tccgccacca cccccgccgc ccccgccacc
       61 gcccggtctg tcccctcggg ctcctgcgcc gccacccgcc ggggccctcc tcccggagcc
      121 cggccagcgc tgcgaggcgg tcagcagcag ccccccgccg cctccctgcg cccagaaccc
      181 cctgcacccc agcccgtccc actccgcgtg caaagagccc gtcgtctacc cctggatgcg
      241 caaagttcac gtgagcacgg taaaccccaa ttacgccggc ggggagccca agcgctctcg
      301 gaccgcctac acgcgccagc aggtcttgga gctggagaag gaatttcact acaaccgcta
      361 cctgacacgg cgccggaggg tggagatcgc ccacgcgctc tgcctctccg agcgccagat
      421 caagatctgg ttccagaacc ggcgcatgaa gtggaaaaaa gaccacaagt tgcccaacac
      481 caagatccgc tcgggtggtg cggcaggctc agccggaggg ccccctggcc ggcccaatgg
      541 aggcccccgc gcgctctagt gcccccgcac gcgggagcca cgaacctcgg ggtgggggtg
      601 ggcagtgagt gcaggggatg gggtgggggg acaggagggg gccctggggc ctgggccccg
      661 gaaaaatcta tctgccctcc cccacacttt atatacgaat aaacgcagaa gagggggagg
      721 ggaagcttcg agcgggatca attccgcccc ccccctaacg ttactggccg aagccgcttg
      781 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc
      841 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc
      901 cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa
      961 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct
     1021 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca
     1081 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca
     1141 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat
     1201 ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc
     1261 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gataatacca tggtgagcaa
     1321 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1381 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1441 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1501 cttcggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     1561 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1621 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1681 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1741 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     1801 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     1861 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     1921 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1981 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga
     2041 ctctagagtc gacctgcagg catgcaagct tcaggtagcc ggctaacgtt aacaaccggt
     2101 acctctagaa ctatagctag catgcgcaaa tttaaagcgc tgatatcgat aaaataaaag
     2161 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag
     2221 ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt
     2281 tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt
     2341 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc
     2401 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     2461 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     2521 tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga
     2581 ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     2641 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     2701 ttcatgggta acagtttctt gaagttggag aacaacattc tgagggtagg agtcgaatat
     2761 taagtaatcc tgactcaatt agccactgtt ttgaatccac atactccaat actcctgaaa
     2821 tagttcatta tggacagcgc agaaagagct ggggagaatt gtgaaattgt tatccgctca
     2881 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     2941 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3001 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3061 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3121 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3181 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3241 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3301 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3361 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3421 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3481 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3541 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3601 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3661 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3721 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3781 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3841 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3901 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3961 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4021 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4081 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4141 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4201 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4261 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4321 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4381 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4441 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4501 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4561 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4621 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4681 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4741 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4801 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4861 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4921 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4981 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     5041 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     5101 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     5161 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     5221 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     5281 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     5341 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     5401 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggcgcaag
     5461 gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat
     5521 acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt
     5581 gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat
     5641 gcgtccggcg tagaggcgat tagtccaatt tgttaaagac aggatatcag tggtccaggc
     5701 tctagttttg actcaacaat atcaccagct gaagcctata gagtacgagc catagataaa
     5761 ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt
     5821 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata actgagaata
     5881 gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca aacaggatat
     5941 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt
     6001 cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga
     6061 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
     6121 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc
     6181 cgatagactg cgtcgcccgg gtacccgtat tcccaataaa gcctcttgct gtttgcatcc
     6241 gaatcgtgga ctcgctgatc cttgggaggg tctcctcaga ttgattgact gcccacctcg
     6301 ggggtctttc atttggaggt tccaccgaga tttggagacc cctgcccagg gaccaccgac
     6361 ccccccgccg ggaggtaagc tggccagcgg tcgtttcgtg tctgtctctg tctttgtgcg
     6421 tgtttgtgcc ggcatctaat gtttgcgcct gcgtctgtac tagttagcta actagctctg
     6481 tatctggcgg acccgtggtg gaactgacga gttctgaaca cccggccgca accctgggag
     6541 acgtcccagg gactttgggg gccgtttttg tggcccgacc tgaggaaggg agtcgatgtg
     6601 gaatccgacc ccgtcaggat atgtggttct ggtaggagac gagaacctaa aacagttccc
     6661 gcctccgtct gaatttttgc tttcggtttg gaaccgaagc cgcgcgtctt gtctgctgca
     6721 gcgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa
     6781 ttagggccag actgttacca ctcccttaag tttgacctta ggtcactgga aagatgtcga
     6841 gcggatcgct cacaaccagt cggtagatgt caagaagaga cgttgggtta ccttctgctc
     6901 tgcagaatgg ccaaccttta acgtcggatg gccgcgagac ggcaccttta accgagacct
     6961 catcacccag gttaagatca aggtcttttc acctggcccg catggacacc cagaccaggt
     7021 cccctacatc gtgacctggg aagccttggc ttttgacccc cctccctggg tcaagccctt
     7081 tgtacaccct aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc
     7141 tcctcgttcg accccgcctc gatcctccct ttatccagcc ctcactcctt ctctaggcgc
     7201 cggaattaat tcgggttagt atattttgtg ggcaattccc agaaattaat ggctatgagt
     7261 tcttttttga tcaactcaaa ctatgtcgac cccaagttcc ctccatgcga ggaatattca
     7321 cagagcgatt acctacccag cgaccactcg cccgggtact acgccggcgg ccagaggcga
     7381 gagagcagct tccagccgga ggcgggcttc gggcggcgcg cggcgtgcac cgtgcagcgc
     7441 tacgc
//
