LOCUS       sequence-337876-        8331 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive overexpression of let-7c with mTagBFP2 as a reporter 
            fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8331)
  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 8331)
  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.mTagBFP2.let-7c
FEATURES             Location/Qualifiers
     source          1..8331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /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    1748..1865
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1923..2324
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      2335..2344
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3521..4109
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3574..3594)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3992..4003)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4181..4414
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4492..4613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4529..4548)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4583..4602
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4653..4788
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4808..4827)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4809..4827)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4837..4854)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4837..4853)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4846..4868)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4995..5450
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5082..5101)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5292..5313
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5476..5580
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5581..5649,5650..6441)
                     /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(5799..5818)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6612..7200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7101..7120
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7354..7371
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7488..7509
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7524..7529,7530..7547,7548..7554)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7562..7578
                     /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     7567..7589
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7586..7602
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7586..7602
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7621..7641
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7623..7641
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        7686..8065
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     enhancer        8122..8313
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(8145..8165)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggacaat tctagatctt gagacaaatg gcagtattca
     1741 tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag
     1801 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
     1861 attttcgggt ttattacagg gacagcagag atccactttg gcgccggctc gagcgagctg
     1921 cagtaacgcc attttgcaag gcatggaaaa ataccaaacc aagaatagag aagttcagat
     1981 caagggcggg tacatgaaaa tagctaacgt tgggccaaac aggatatctg cggtgagcag
     2041 tttcggcccc ggcccggggc caagaacaga tggtcaccgc agtttcggcc ccggcccgag
     2101 gccaagaaca gatggtcccc agatatggcc caaccctcag cagtttctta agacccatca
     2161 gatgtttcca ggctccccca aggacctgaa atgaccctgc gccttatttg aattaaccaa
     2221 tcagcctgct tctcgcttct gttcgcgcgc ttctgcttcc cgagctctat aaaagagctc
     2281 acaacccctc actcggcgcg ccagtcctcc gacagactga gtcggatcgc ggccgccacc
     2341 atgggaagcg agctgattaa ggagaacatg cacatgaagc tctacatgga gggcaccgtg
     2401 gacaaccatc acttcaagtg cacatccgag ggcgaaggca agccctacga gggcacccag
     2461 accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga catcctggct
     2521 actagcttcc tctacggcag caagaccttc atcaaccaca cccagggcat ccccgacttc
     2581 ttcaagcagt ccttccctga gggcttcaca tgggagagag tcaccacata cgaagacggg
     2641 ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat ctacaacgtc
     2701 aagatcagag gggtgaactt cacatccaac ggccctgtga tgcagaagaa aacactcggc
     2761 tgggaggcct tcaccgagac cctgtacccc gctgacggcg gcctggaagg cagaaacgac
     2821 atggccctga agctcgtggg cgggagccat ctgatcgcaa acatcaagac cacatataga
     2881 tccaagaaac ccgctaagaa cctcaagatg cctggcgtct actatgtgga ctacagactg
     2941 gaaagaatca aggaggccaa caacgagacc tacgtcgagc agcacgaggt ggcagtggcc
     3001 agatactgcg acctccctag caaactgggg cacaagctgt acaagtgagg atccgcaggt
     3061 tagatggtca gaagacattt tacgtgacct atgctggaaa cccaagtgtt taggttagtc
     3121 ctatgagaaa cagttagcag caaagtttac tggtaaaatg ttagaatatt atatctatat
     3181 ccttgccaag cccttaggtg tatggctgcc atatttggag gagctgactg aagatatgat
     3241 aaggagtttg aagcaacatt ggaagctgtg tgcatccggg ttgaggtagt aggttgtatg
     3301 gtttagagtt acaccctggg agttaactgt acaaccttct agctttcctt ggagcacact
     3361 tgagccgtcg aggaattctt catcacttta acctgattga gccaatttgt gtgcaagaag
     3421 gtaatgtgtc atgagtatct tggatcattg atttgggcct ttattacatt aaaatgatca
     3481 tttcagccct attaccggta acgcgtcgcg ttaagtcgac aatcaacctc tggattacaa
     3541 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3601 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3661 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3721 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3781 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3841 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3901 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     3961 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     4021 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     4081 tcggatctcc ctttgggccg cctccccgcg tcgactttaa gaccaatgac ttacaaggca
     4141 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     4201 caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     4261 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     4321 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     4381 agaccctttt agtcagtgtg gaaaatctct agcagtacgt atagtagttc atgtcatctt
     4441 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt
     4501 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4561 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4621 tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4681 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4741 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc
     4801 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg
     4861 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     4921 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     4981 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     5041 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     5101 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     5161 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     5221 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     5281 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     5341 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     5401 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     5461 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5521 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5581 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5641 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5701 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5761 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5821 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5881 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5941 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6001 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6061 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     6121 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     6181 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6241 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     6301 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6361 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6421 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6481 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6541 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6601 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6661 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6721 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6781 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6841 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6901 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6961 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7021 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7081 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     7141 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7201 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     7261 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     7321 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     7381 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     7441 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     7501 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     7561 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
     7621 gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcggtacc gtcgacctgc
     7681 aggttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     7741 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     7801 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     7861 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     7921 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     7981 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     8041 cgtattagtc atcgctatta ccatggcgcg ccatgcaggg cctgaaataa cctctgaaag
     8101 aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     8161 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     8221 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     8281 catgcatctc aattagtcag caaccatagt cccttaagag gcgcgcctta a
//
