LOCUS       sequence-338325-        8267 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Constitutive overexpression of miR-125b-2 with mTagBFP2 as a 
            reporter fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8267)
  AUTHORS   Alejo-Valle O, Weigert K, Bhayadia R, Ng M, Issa H, Beyer C, Emmrich
            S, Schuschel K, Ihling C, Sinz A, Zimmermann M, Wickenhauser C, 
            Flasinski M, Regenyi EM, Labuhn M, Reinhardt D, Yaspo ML, Heckl D, 
            Klusmann JH
  TITLE     The megakaryocytic transcription factor ARID3A suppresses leukemia 
            pathogenesis.
  JOURNAL   Blood. 2021 Sep 27. pii: S0006-4971(21)01649-9. doi: 
            10.1182/blood.2021012231.
  PUBMED    34570885
REFERENCE   2  (bases 1 to 8267)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Blood. 2021
            Sep 27. pii: S0006-4971(21)01649-9. doi: 10.1182/blood.2021012231."
COMMENT     Sequence Label: SIN40C.SFFV.mTagBFP2.miR-125b-2
FEATURES             Location/Qualifiers
     source          1..8267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        89..280
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     237..257
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     enhancer        337..716
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(761..781)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(761..779)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(800..816)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(800..816)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(813..835)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    824..840
                     /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(848..854,855..872,873..878))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    893..914
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1031..1048)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1202..1790)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1282..1301)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1961..2752,2753..2821))
                     /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     2584..2603
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2822..2926)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2952..3407)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3089..3110)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3301..3320
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3534..3556
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3548..3565
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3549..3565
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3575..3594
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3575..3593
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      3614..3749
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3789..3910
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3800..3819)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3854..3873
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             3988..4221
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    4293..4881
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             4399..4410
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4808..4828
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     regulatory      5994..6003
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6014..6415)
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     misc_feature    6473..6590
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(6803..6847)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7032..7265
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    7758..7883
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             7930..8110
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(join(8111..8267,1..70))
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 ccttgtaagg catgttgcta actcatcgtt accatgttgc aagactacaa gagtattgca
       61 taagactaca ttaaggcgcg cctcttaagg gactatggtt gctgactaat tgagatgcat
      121 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta
      181 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc
      241 taactgacac acattccaca gctggttctt tccgcctcag aaggtaccta accaagttcc
      301 tctttcagag gttatttcag gccctgcatg gcgcgccatg gtaatagcga tgactaatac
      361 gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc
      421 gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg
      481 tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc
      541 tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg
      601 cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct
      661 atgaactaat gaccccgtaa ttgattacta ttaataacta gtcaataatc aatgtcaacc
      721 tgcaggtcga cggtaccgct tgcagctcca gcttttgttc cctttagtga gggttaattg
      781 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
      841 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
      901 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
      961 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     1021 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     1081 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     1141 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1201 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1261 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1321 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1381 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1441 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1501 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1561 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1621 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1681 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1741 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1801 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1861 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1921 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1981 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     2041 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     2101 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     2161 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2221 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2281 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2341 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     2401 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     2461 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     2521 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     2581 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     2641 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     2701 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     2761 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2821 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     2881 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2941 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     3001 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3061 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     3121 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     3181 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3241 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3301 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     3361 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     3421 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     3481 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     3541 acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt
     3601 gggtacgtcc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc
     3661 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
     3721 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggat agctagagcc
     3781 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3841 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3901 taaacaagtt cctctcactc tctgatattc atttctttgc aagttataaa tactgaataa
     3961 taagatgaca tgaactacta tacgtactgc tagagatttt ccacactgac taaaagggtc
     4021 tgagggatct ctagttacca gagtcacaca acagacgggc acacactact tgaagcactc
     4081 aaggcaagct ttattgaggc ttaagcagtg ggttccctag ttagccagag agctcccagg
     4141 ctcagatctg gtctaaccag agagacccag tacaagcaaa aagcagatct tgtcttcgtt
     4201 gggagtgaat tagcccttcc agtcccccct tttcttttaa aaagtggcta agatctacag
     4261 ctgccttgta agtcattggt cttaaagtcg acgcggggag gcggcccaaa gggagatccg
     4321 actcgtctga gggcgaaggc gaagacgcgg aagaggccgc agagccggca gcaggccgcg
     4381 ggaaggaagg tccgctggat tgagggccga agggacgtag cagaaggacg tcccgcgcag
     4441 aatccaggtg gcaacacagg cgagcagcca aggaaaggac gatgatttcc ccgacaacac
     4501 cacggaattg tcagtgccca acagccgagc ccctgtccag cagcgggcaa ggcaggcggc
     4561 gatgagttcc gccgtggcaa tagggagggg gaaagcgaaa gtcccggaaa ggagctgaca
     4621 ggtggtggca atgccccaac cagtgggggt tgcgtcagca aacacagtgc acaccacgcc
     4681 acgttgcctg acaacgggcc acaactcctc ataaagagac agcaaccagg atttatacaa
     4741 ggaggagaaa atgaaagcca tacgggaagc aatagcatga tacaaaggca ttaaagcagc
     4801 gtatccacat agcgtaaaag gagcaacata gttaagaata ccagtcaatc tttcacaaat
     4861 tttgtaatcc agaggttgat tgtcgactta acgcgacgcg ttaccggtgg tgaaaacgtt
     4921 gctgataaag aaaagcattg ttcttttctc ctaggcagaa tctatgtatg ttcttttgtt
     4981 ctggtggtaa ctgcagacaa tcaataaggt ccaaagctgc tggttcccct ccgcctaggt
     5041 cccaagagcc tgacttgtga tgttactaaa atacctcaca agttagggtc tcagggacta
     5101 ggaaaagtct ggtttgatgc ggtagaattc ttcttccaga aataaggata gcttagaaat
     5161 tacagttggg ttcacttaga cttttacctc taaatactat ttaaaatact tcagtagaaa
     5221 tatatatgat atataaaagt taggatgagc tgagtaatca cgacgaccat ttatagacta
     5281 attggatcct cacttgtaca gcttgtgccc cagtttgcta gggaggtcgc agtatctggc
     5341 cactgccacc tcgtgctgct cgacgtaggt ctcgttgttg gcctccttga ttctttccag
     5401 tctgtagtcc acatagtaga cgccaggcat cttgaggttc ttagcgggtt tcttggatct
     5461 atatgtggtc ttgatgtttg cgatcagatg gctcccgccc acgagcttca gggccatgtc
     5521 gtttctgcct tccaggccgc cgtcagcggg gtacagggtc tcggtgaagg cctcccagcc
     5581 gagtgttttc ttctgcatca cagggccgtt ggatgtgaag ttcacccctc tgatcttgac
     5641 gttgtagatg aggcagccgt cctggaggct ggtgtcctgg gtagcggtca gcacgccccc
     5701 gtcttcgtat gtggtgactc tctcccatgt gaagccctca gggaaggact gcttgaagaa
     5761 gtcggggatg ccctgggtgt ggttgatgaa ggtcttgctg ccgtagagga agctagtagc
     5821 caggatgtcg aaggcgaagg ggagagggcc gccctcgacc accttgattc tcatggtctg
     5881 ggtgccctcg tagggcttgc cttcgccctc ggatgtgcac ttgaagtgat ggttgtccac
     5941 ggtgccctcc atgtagagct tcatgtgcat gttctcctta atcagctcgc ttcccatggt
     6001 ggcggccgcg atccgactca gtctgtcgga ggactggcgc gccgagtgag gggttgtgag
     6061 ctcttttata gagctcggga agcagaagcg cgcgaacaga agcgagaagc aggctgattg
     6121 gttaattcaa ataaggcgca gggtcatttc aggtccttgg gggagcctgg aaacatctga
     6181 tgggtcttaa gaaactgctg agggttgggc catatctggg gaccatctgt tcttggcctc
     6241 gggccggggc cgaaactgcg gtgaccatct gttcttggcc ccgggccggg gccgaaactg
     6301 ctcaccgcag atatcctgtt tggcccaacg ttagctattt tcatgtaccc gcccttgatc
     6361 tgaacttctc tattcttggt ttggtatttt tccatgcctt gcaaaatggc gttactgcag
     6421 ctcgctcgag ccggcgccaa agtggatctc tgctgtccct gtaataaacc cgaaaatttt
     6481 gaatttttgt aatttgtttt tgtaattctt tagtttgtat gtctgttgct attatgtcta
     6541 ctattctttc ccctgcactg taccccccaa tccccccttt tcttttaaaa ttgtggatga
     6601 atactgccat ttgtctcaag atctagaatt gtcccctcgg ggttgggagg tgggtctgaa
     6661 acgataatgg tgaatatccc tgcctaactc tattcactat agaaagtaca gcaaaaacta
     6721 ttcttaaacc taccaagcct cctactatca ttatgaataa ttttatatac cacagccaat
     6781 ttgttatgtt aaaccaattc cacaaacttg cccatttatc taattccaat aattcttgtt
     6841 cattcttttc ttgctggttt tgcgattctt caattaagga gtgtattaag cttgtgtaat
     6901 tgttaatttc tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt tccagagatt
     6961 tattactcca actagcattc caaggcacag cagtggtgca aatgagtttt ccagagcaac
     7021 cccaaatccc caggagctgt tgatccttta ggtatctttc cacagccagg attcttgcct
     7081 ggagctgctt gatgccccag actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc
     7141 tcagcaaatt gttctgctgc tgcactatac cagacaataa ttgtctggcc tgtaccgtca
     7201 gcgtcattga cgctgcgccc atagtgcttc ctgctgctcc caagaaccca aggaacaaag
     7261 ctcctattcc cactgctctt ttttctctct gcaccactct tctctttgcc ttggtgggtg
     7321 ctactcctaa tggttcaatt tttactactt tatatttata taattcactt ctccaattgt
     7381 ccctcatatc tcctcctcca ggtctgaaga tcagcggccg cttgctgtgc ggtggtctta
     7441 cttttgtttt gctcttcctc tatcttgtct aaagcttcct tggtgtcttt tatctctatc
     7501 ctttgatgca cacaatagag ggttgctact gtattatata atgatctaag ttcttctgat
     7561 cctgtctgaa gggatggttg tagctgtccc agtatttgtc tacagccttc tgatgtttct
     7621 aacaggccag gattaactgc gaatcgttct agctccctgc ttgcccatac tatatgtttt
     7681 aatttatatt ttttctttcc ccctggcctt aaccgaattt tttcccatcg cgatctaatt
     7741 ctcccccgct taatactgac gctctcgcac ccatctctct ccttctagcc tccgctagtc
     7801 aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca
     7861 agccgagtcc tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa gtccctgttc
     7921 gggcgccact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
     7981 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
     8041 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
     8101 agagagaccc gtttatgtat cgagctaggc acttaaatac aatatctctg caatgcggca
     8161 attcagtggt tcgtccaatc catgtcagac ccgtctgttg ccttcctaat aaggcacgat
     8221 cgtaccacct tacttccacc aatcggcatg cacggtgctt tttctct
//
