LOCUS       sequence-450571-        8155 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The most commonly used plasmid for the generation of ER-Hoxb8 
            conditionally immortalized myeloid cell lines..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8155)
  AUTHORS   Wang GG, Calvo KR, Pasillas MP, Sykes DB, Hacker H, Kamps MP
  TITLE     Quantitative production of macrophages or neutrophils ex vivo using 
            conditional Hoxb8.
  JOURNAL   Nat Methods. 2006 Apr;3(4):287-93.
  PUBMED    16554834
REFERENCE   2  (bases 1 to 8155)
  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"; journalName: "Nat 
            Methods."; date: "2006-04"; volume: "3(4)"; pages: "287-93"
COMMENT     Sequence Label: MSCVneo-HA-ER-Hoxb8
FEATURES             Location/Qualifiers
     source          1..8155
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1411..1420
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1423..1449
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1459..2394
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             1507..2397
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     promoter        3157..3656
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3192..3215)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3589..3608
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3670..4473
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3733..3752)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4343..4362
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             4548..5062
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5224..5240
                     /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(5248..5254,5255..5272,5273..5278))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5293..5314
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5431..5448)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5602..6190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5682..5701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6361..7152,7153..7221))
                     /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     6984..7003
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7222..7326)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7394..7412
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7450..7472)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7572..7591
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7785..7807
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7935..7954
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8154..8155,1..515)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaattc gccaccatgg gatacccata cgatgttccg
     1441 gattacgcta cgcgttctgc tggagacatg agagctgcca acctttggcc aagcccgctc
     1501 atgatcaaac gctctaagaa gaacagcctg gccttgtccc tgacggccga ccagatggtc
     1561 agtgccttgt tggatgctga gccccccata ctctattccg agtatgatcc taccagaccc
     1621 ttcagtgaag cttcgatgat gggcttactg accaacctgg cagacaggga gctggttcac
     1681 atgatcaact gggcgaagag ggtgccaggc tttgtggatt tgaccctcca tgatcaggtc
     1741 caccttctag aatgtgcctg gctagagatc ctgatgattg gtctcgtctg gcgctccatg
     1801 gagcacccag tgaagctact gtttgctcct aacttgctct tggacaggaa ccagggaaaa
     1861 tgtgtagagg gcatggtgga gatcttcgac atgctgctgg ctacatcatc tcggttccgc
     1921 atgatgaatc tgcagggaga ggagtttgtg tgcctcaaat ctattatttt gcttaattct
     1981 ggagtgtaca catttctgtc cagcaccctg aagtctctgg aagagaagga ccatatccac
     2041 cgagtcctgg acaagatcac agacactttg atccacctga tggccaaggc aggcctgacc
     2101 ctgcagcagc agcaccagcg gctggcccag ctcctcctca tcctctccca catcaggcac
     2161 atgagtaaca aaggcatgga gcatctgtac agcatgaagt gcaagaacgt ggtgcccctc
     2221 tatgacctgc tgctggagat gctggacgcc caccgcctac atgcgcccac tagccgtgga
     2281 ggggcatccg tggaggagac ggaccaaagc cacttggcca ctgcgggctc tacttcatcg
     2341 cattccttgc aaaagtatta catcacgggg gaggcagagg gtttccctgc cacagtcacg
     2401 cgtggaagct cttatttcgt caactcactg ttctccaaat acaaaaccgg ggagtccctg
     2461 cgccccaatt attatgactg cggcttcgcc caggacctgg gcggccgacc caccgtggtg
     2521 tacggtccca gcagcggcgg cagcttccag cacccttcgc aaatccagga gttctaccac
     2581 gggccatcgt cgctgtccac agctccctac cagcagaacc cgtgcgccgt ggcgtgccac
     2641 ggcgaccccg gcaacttcta cggctacgac cctctgcagc gccagagcct gttcggtgcg
     2701 caggatccag acctggtgca gtacgcagac tgcaagctcg cggcagccag cggcctgggc
     2761 gaggaggccg aggggtctga gcagagcccg tcgcccacac agctctttcc ctggatgcgc
     2821 cctcaagccg ccggacgcag gcgaggccgc cagacctaca gtcgctacca gaccctggag
     2881 ctggagaagg agttcctatt taatccctat ctgactcgca agcggaggat cgaggtatcg
     2941 cacgcgctgg gactgacaga gagacaggtc aaaatctggt tccagaatcg gagaatgaag
     3001 tggaaaaagg agaacaacaa agacaagttt cccagcagta aatgcgagca ggaggagctg
     3061 gagaaagaga agctggagcg ggcaccagag accgccgagc agggcgatgc gcagaagggt
     3121 gacaagaagt agtaactcga gagatctaat tctaccgggt aggggaggcg cttttcccaa
     3181 ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc
     3241 tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg
     3301 gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc
     3361 tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg
     3421 acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt
     3481 gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg
     3541 gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc
     3601 ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct
     3661 gcagccaata tgggatcggc cattgaacaa gatggattgc acgcaggttc tccggccgct
     3721 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3781 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3841 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3901 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3961 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4021 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4081 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4141 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4201 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4261 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4321 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4381 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4441 gccttctatc gccttcttga cgagttcttc tgaggggatc cgtcgacctg cagccaagct
     4501 tatcgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     4561 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata
     4621 actgagaata gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca
     4681 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     4741 cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     4801 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     4861 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc
     4921 gccagtcctc cgatagactg cgtcgcccgg gtacccgtgt atccaataaa ccctcttgca
     4981 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta
     5041 cccgtcagcg ggggtctttc atgggtaaca gtttcttgaa gttggagaac aacattctga
     5101 gggtaggagt cgaatattaa gtaatcctga ctcaattagc cactgttttg aatccacata
     5161 ctccaatact cctgaaatag ttcattatgg acagcgcaga aagagctggg gagaattgtg
     5221 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5281 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5341 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5401 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5461 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5521 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5581 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5641 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5701 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5761 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5821 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5881 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5941 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6001 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6061 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6121 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6181 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6241 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6301 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6361 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6421 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6481 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6541 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6601 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6661 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6721 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6781 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6841 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6901 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6961 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7021 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7081 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7141 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7201 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7261 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7321 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     7381 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     7441 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     7501 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     7561 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     7621 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     7681 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     7741 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     7801 taaaacgacg gcgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     7861 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     7921 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     7981 gtgatgccgg ccacgatgcg tccggcgtag aggcgattag tccaatttgt taaagacagg
     8041 atatcagtgg tccaggctct agttttgact caacaatatc accagctgaa gcctatagag
     8101 tacgagccat agataaaata aaagatttta tttagtctcc agaaaaaggg gggaa
//
