LOCUS       sequence-337881-        8266 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive overexpression of miR-125b-2 with dTomato as a reporter
            fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8266)
  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 8266)
  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: "Blood. 2021
            Sep 27. pii: S0006-4971(21)01649-9. doi: 10.1182/blood.2021012231."
COMMENT     Sequence Label: SIN40C.SFFV.dTomato.miR-125b-2
FEATURES             Location/Qualifiers
     source          1..8266
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             10..190
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    237..362
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    855..1088
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1273..1317
                     /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    1530..1647
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1705..2112
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      2112..2121
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2118..2819
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(2251..2269)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2519..2538
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2713..2736
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    3238..3826
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3291..3311)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3709..3720)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3898..4131
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4209..4330
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4246..4265)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4300..4319
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4370..4505
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4525..4544)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4526..4544)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4554..4571)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4554..4570)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4563..4585)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4712..5167
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4799..4818)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5009..5030
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5193..5297
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5298..5366,5367..6158)
                     /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     complement(5516..5535)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6329..6917
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6818..6837
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7071..7088
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7205..7226
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7241..7246,7247..7264,7265..7271)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7279..7295
                     /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)."
     primer_bind     7284..7306
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7303..7319
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7303..7319
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7338..7358
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7340..7358
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        7403..7782
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     enhancer        7839..8030
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(7862..7882)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(8049..8266,1..9)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
       61 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
      121 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
      181 atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct
      241 ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg
      301 tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg
      361 tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg
      421 gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg
      481 cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac
      541 aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc
      601 tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag
      661 aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct
      721 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
      781 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
      841 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
      901 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
      961 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
     1021 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
     1081 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
     1141 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     1201 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     1261 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     1321 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     1381 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     1441 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggaca attctagatc
     1501 ttgagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac
     1561 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa
     1621 aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag agatccactt
     1681 tggcgccggc tcgagcgagc tgcagtaacg ccattttgca aggcatggaa aaataccaaa
     1741 ccaagaatag agaagttcag atcaagggcg ggtacatgaa aatagctaac gttgggccaa
     1801 acaggatatc tgcggtgagc agtttcggcc ccggcccggg gccaagaaca gatggtcacc
     1861 gcagtttcgg ccccggcccg aggccaagaa cagatggtcc ccagatatgg cccaaccctc
     1921 agcagtttct taagacccat cagatgtttc caggctcccc caaggacctg aaatgaccct
     1981 gcgccttatt tgaattaacc aatcagcctg cttctcgctt ctgttcgcgc gcttctgctt
     2041 cccgagctct ataaaagagc tcacaacccc tcactcggcg cgccagtcct ccgacagact
     2101 gagtcggatc cgccaccatg gtgagcaagg gcgaggaggt catcaaagag ttcatgcgct
     2161 tcaaggtgcg catggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg
     2221 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc
     2281 tgcccttcgc ctgggacatc ctgtcccccc agttcatgta cggctccaag gcgtacgtga
     2341 agcaccccgc cgacatcccc gattacaaga agctgtcctt ccccgagggc ttcaagtggg
     2401 agcgcgtgat gaacttcgag gacggcggtc tggtgaccgt gacccaggac tcctccctgc
     2461 aggacggcac gctgatctac aaggtgaaga tgcgcggcac caacttcccc cccgacggcc
     2521 ccgtaatgca gaagaagacc atgggctggg aggcctccac cgagcgcctg tacccccgcg
     2581 acggcgtgct gaagggcgag atccaccagg ccctgaagct gaaggacggc ggccactacc
     2641 tggtggagtt caagaccatc tacatggcca agaagcccgt gcaactgccc ggctactact
     2701 acgtggacac caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     2761 acgagcgctc cgagggccgc caccacctgt tcctgtacgg catggacgag ctgtacaagt
     2821 gagctagccg tctcagatcc aattagtcta taaatggtcg tcgtgattac tcagctcatc
     2881 ctaactttta tatatcatat atatttctac tgaagtattt taaatagtat ttagaggtaa
     2941 aagtctaagt gaacccaact gtaatttcta agctatcctt atttctggaa gaagaattct
     3001 accgcatcaa accagacttt tcctagtccc tgagacccta acttgtgagg tattttagta
     3061 acatcacaag tcaggctctt gggacctagg cggaggggaa ccagcagctt tggaccttat
     3121 tgattgtctg cagttaccac cagaacaaaa gaacatacat agattctgcc taggagaaaa
     3181 gaacaatgct tttctttatc agcaacgttt tcaccaccgg taacgcgtta agtcgacaat
     3241 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     3301 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     3361 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     3421 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3481 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     3541 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3601 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     3661 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3721 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3781 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac
     3841 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     3901 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     3961 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     4021 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     4081 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtacgtata
     4141 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     4201 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4261 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4321 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct
     4381 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4441 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     4501 aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
     4561 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     4621 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     4681 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     4741 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4801 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4861 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4921 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4981 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5041 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5101 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     5161 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5221 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5281 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5341 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5401 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5461 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5521 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5581 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5641 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5701 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5761 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5821 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5881 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5941 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6001 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6061 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6121 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6181 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6241 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6301 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6361 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6421 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6481 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6541 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6601 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6661 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6721 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6781 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6841 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6901 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6961 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7021 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     7081 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     7141 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     7201 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     7261 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     7321 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag
     7381 cggtaccgtc gacctgcagg ttgacattga ttattgacta gttattaata gtaatcaatt
     7441 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     7501 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     7561 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     7621 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     7681 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     7741 acttggcagt acatctacgt attagtcatc gctattacca tggcgcgcca tgcagggcct
     7801 gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg
     7861 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     7921 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     7981 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc ttaagaggcg
     8041 cgccttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta
     8101 gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg
     8161 gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac
     8221 tgaattgccg cattgcagag atattgtatt taagtgccta gctcga
//
