LOCUS       sequence-91470-e        6141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MSCV based expression vector - co-expresses mCherry and human 
            miR-125b-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6141)
  AUTHORS   So AY, Sookram R, Chaudhuri AA, Minisandram A, Cheng D, Xie C, Lim 
            EL, Flores YG, Jiang S, Kim JT, Keown C, Ramakrishnan P, Baltimore D
  TITLE     Dual mechanisms by which miR-125b represses IRF4 to induce myeloid 
            and B-cell leukemias.
  JOURNAL   Blood. 2014 Aug 28;124(9):1502-12. doi: 
            10.1182/blood-2014-02-553842. Epub 2014 Jul 8.
  PUBMED    25006123
REFERENCE   2  (bases 1 to 6141)
  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: 
            "10.1182/blood-2014-02-553842"; journalName: "Blood"; date: 
            "2014-08-28- 28"; volume: "124"; issue: "9"; pages: "1502-12"
COMMENT     Sequence Label: pmCherry-miR-125b-1
FEATURES             Location/Qualifiers
     source          1..6141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    580..921
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             988..1404
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1334..1356
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1372..1388
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1432..1441
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1438..2148
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1583..1601)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(3249..3265)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3249..3265)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3262..3284)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3273..3289
                     /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(3297..3303,3304..3321,3322..3327))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3342..3363
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3480..3497)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3651..4239)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3731..3750)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4410..5201,5202..5270))
                     /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     5033..5052
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5271..5375)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5443..5461
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5499..5521)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5621..5640
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5834..5856
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5848..5865
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5849..5865
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6064..6083
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
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 gaaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
      301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
      361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat
      421 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc
      481 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag
      541 acccctgcct agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc
      601 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg
      661 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga
      721 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg
      781 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga
      841 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga
      901 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact
      961 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc
     1021 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag
     1081 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga
     1141 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc
     1201 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac
     1261 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
     1321 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
     1381 gccctcactc cttctctagg cgccggaatt agatccccgg gtaccggtgc cgccaccatg
     1441 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     1501 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     1561 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     1621 gcctgggaca tcctgtcccc tcaattcatg tatggctcca aggcctacgt gaagcacccc
     1681 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     1741 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctcctt acaagacggc
     1801 gagttcatct acaaagtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     1861 caaaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
     1921 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     1981 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
     2041 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     2101 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaagc ggccgcgagt
     2161 tttctctgat gtactcgtga tcgtatgtgt atgtgcgtga ttgtatatgc gcccccagat
     2221 actgcgtatg tgtgtatata tgtattaggc ttaaacggaa tctcaatttt gtgaaggaaa
     2281 ggagcttaga gaagaaatac cataccacct gtttgttgca tcttagttat gaacctcgaa
     2341 cagaaattgc ctgtcattct tgttttgctt tgctttgtct caagaaagaa aacattgttg
     2401 cgctcctctc agtccctgag accctaactt gtgatgttta ccgtttaaat ccacgggtta
     2461 ggctcttggg agctgcgagt cgtgcttttg catcctggaa atttggtgga attttattct
     2521 ttaaagcaaa aacaaaagaa aagaaagttt gtctgaggtg attgagtata cctctgaggt
     2581 tttcattgtt agatgggatc aggtgaccag agagtggcag ctcctggatt tctgttctcg
     2641 aggttaacga atttcgacct gcagccaagc ttatcgataa aataaaagat tttatttagt
     2701 ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta
     2761 acgccatttt gcaaggcatg gaaaatacat aactgagaat agagaagttc agatcaaggt
     2821 taggaacaga gagacagcag aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     2881 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tcccgccctc agcagtttct
     2941 agagaaccat cagatgtttc cagggtgccc caaggacctg aaaatgaccc tgtgccttat
     3001 ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc
     3061 aataaaagag cccacaaccc ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc
     3121 gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt
     3181 ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt tcagtattcg
     3241 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3301 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3361 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3421 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3481 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3541 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3601 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3661 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3721 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3781 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3841 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3901 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3961 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4021 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4081 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4141 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4201 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4261 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4321 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4381 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4441 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4501 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4561 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4621 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4681 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4741 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4801 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4861 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4921 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4981 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5041 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5101 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5161 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5221 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5281 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5341 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5401 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5461 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     5521 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     5581 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     5641 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     5701 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     5761 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     5821 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccac gctctccctt
     5881 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     5941 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     6001 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     6061 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     6121 cacgatgcgt ccggcgtaga g
//
