LOCUS       sequence-338319-        8362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive overexpression of miR-ctrl with mTagBFP2 as a reporter 
            fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8362)
  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 8362)
  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.miR-ctrl
FEATURES             Location/Qualifiers
     source          1..8362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    239..360
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(276..295)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     330..349
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      400..535
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(555..574)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(556..574)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(584..601)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(584..600)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(593..615)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      742..1197
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(829..848)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1039..1060
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1223..1327
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1328..1396,1397..2188)
                     /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(1546..1565)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2359..2947
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2848..2867
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3101..3118
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3235..3256
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3271..3276,3277..3294,3295..3301)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3309..3325
                     /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     3314..3336
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3333..3349
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3333..3349
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3368..3388
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3370..3388
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        3433..3812
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     enhancer        3869..4060
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3892..3912)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4079..4305
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4306..4486
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4533..4658
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5151..5384
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5569..5613
                     /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    5826..5943
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6001..6402
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      6413..6422
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    7630..8218
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7683..7703)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8101..8112)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             join(8290..8362,1..161)
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
ORIGIN
        1 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
       61 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
      121 atccctcaga cccttttagt cagtgtggaa aatctctagc agtacgtata gtagttcatg
      181 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
      241 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
      301 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
      361 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc
      421 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
      481 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg
      541 acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
      601 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
      661 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
      721 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
      781 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
      841 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
      901 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
      961 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     1021 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     1081 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     1141 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     1201 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1261 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1321 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1381 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1441 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1501 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1561 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1621 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1681 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1741 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1801 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1861 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1921 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1981 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2041 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2101 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2161 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2221 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2281 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2341 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2401 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2461 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2521 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2581 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2641 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2701 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2761 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2821 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2881 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2941 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3001 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3061 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3121 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     3181 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     3241 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     3301 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     3361 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag cggtaccgtc
     3421 gacctgcagg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     3481 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     3541 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     3601 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     3661 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     3721 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     3781 acatctacgt attagtcatc gctattacca tggcgcgcca tgcagggcct gaaataacct
     3841 ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt
     3901 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3961 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4021 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc ttaagaggcg cgccttaatg
     4081 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
     4141 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
     4201 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
     4261 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
     4321 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     4381 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     4441 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     4501 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     4561 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     4621 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     4681 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     4741 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     4801 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     4861 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     4921 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     4981 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     5041 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     5101 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     5161 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     5221 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     5281 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     5341 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     5401 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     5461 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     5521 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     5581 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     5641 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     5701 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     5761 acccacctcc caaccccgag gggacaattc tagatcttga gacaaatggc agtattcatc
     5821 cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac
     5881 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat
     5941 tttcgggttt attacaggga cagcagagat ccactttggc gccggctcga gcgagctgca
     6001 gtaacgccat tttgcaaggc atggaaaaat accaaaccaa gaatagagaa gttcagatca
     6061 agggcgggta catgaaaata gctaacgttg ggccaaacag gatatctgcg gtgagcagtt
     6121 tcggccccgg cccggggcca agaacagatg gtcaccgcag tttcggcccc ggcccgaggc
     6181 caagaacaga tggtccccag atatggccca accctcagca gtttcttaag acccatcaga
     6241 tgtttccagg ctcccccaag gacctgaaat gaccctgcgc cttatttgaa ttaaccaatc
     6301 agcctgcttc tcgcttctgt tcgcgcgctt ctgcttcccg agctctataa aagagctcac
     6361 aacccctcac tcggcgcgcc agtcctccga cagactgagt cggatcgcgg ccgccaccat
     6421 gggaagcgag ctgattaagg agaacatgca catgaagctc tacatggagg gcaccgtgga
     6481 caaccatcac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg gcacccagac
     6541 catgagaatc aaggtggtcg agggcggccc tctccccttc gccttcgaca tcctggctac
     6601 tagcttcctc tacggcagca agaccttcat caaccacacc cagggcatcc ccgacttctt
     6661 caagcagtcc ttccctgagg gcttcacatg ggagagagtc accacatacg aagacggggg
     6721 cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct acaacgtcaa
     6781 gatcagaggg gtgaacttca catccaacgg ccctgtgatg cagaagaaaa cactcggctg
     6841 ggaggccttc accgagaccc tgtaccccgc tgacggcggc ctggaaggca gaaacgacat
     6901 ggccctgaag ctcgtgggcg ggagccatct gatcgcaaac atcaagacca catatagatc
     6961 caagaaaccc gctaagaacc tcaagatgcc tggcgtctac tatgtggact acagactgga
     7021 aagaatcaag gaggccaaca acgagaccta cgtcgagcag cacgaggtgg cagtggccag
     7081 atactgcgac ctccctagca aactggggca caagctgtac aagtgaggat ccgctagctg
     7141 catccttaga actcagcatt taaaacattt atactaaagg atcaatatta tttttgactc
     7201 ttaattgcat cagatattta caacaaattc tatattaata gggggcccat gcaagatgtt
     7261 gcccattggc ataaacccgt agatccgatc ttgtggtgaa gtggaccgca caagctcgct
     7321 tctatgggtc tgtgtcagtg tggtaatctg acaaaatgct atacacagtg ccgttcaaca
     7381 atagttcagt aaaatcctgt taaactccag ttgattatat acttttgaag tcattatatt
     7441 ttcctttgtt tttaatgttt attttaatca ttgtctgtct tacaaggcag gcttcaattc
     7501 tcaacaactt ggaagcgttt atatcacacc cattcaagtt cgattccatg tacagtaaat
     7561 tgcataagaa agttgaacct ttatagcagg gtttggacca ggaccggtaa cgcgtcgcgt
     7621 taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     7681 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     7741 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     7801 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     7861 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     7921 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     7981 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     8041 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     8101 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     8161 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt
     8221 cgactttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     8281 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
     8341 ctgggtctct ctggttagac ca
//
