LOCUS       sequence-222786-        5761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  miR-30a RNAi Expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5761)
  TITLE     Mmu-miR-30a Marc Beltra plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5761)
  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: 
            "Unpublished"
COMMENT     Sequence Label: mmu-miR-30a-5p Block-iT design (BLK)
FEATURES             Location/Qualifiers
     source          1..5761
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        74..453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        454..657
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     608..628
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        702..720
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    750..774
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(783..785,786..788,789..1502)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(828..849)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1089..1108)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1436..1457
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     ncRNA           1562..1588
                     /label=5' miR-155
                     /note="sequence upstream of the precursor of mouse miR-155 
                     microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     ncRNA           1654..1695
                     /label=3' miR-155
                     /note="sequence downstream of the precursor of mouse 
                     miR-155 microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     protein_bind    complement(1724..1748)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1790..1809)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1834..1882
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        complement(2053..2071)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2076..2092)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2076..2092)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      2160..2588
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2247..2266)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2457..2478
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2597..2617)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2602..2931
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2782..2917
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2844..2863
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2979..3026
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3045..3443
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3601..3722
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3638..3657)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3692..3711
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3757..3774)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3928..4516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4008..4027)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4612..5403)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
       61 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      121 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      181 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      241 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      301 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      361 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      421 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      481 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      541 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      601 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
      661 tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg
      721 gagtcccaag ctggctagtt aagctatcaa caagtttgta caaaaaagca ggctttaaaa
      781 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
      841 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      901 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
      961 ccctcgtgac caccttcacc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga
     1021 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     1081 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1141 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1201 acaagctgga gtacaactac aacagccaca aggtctatat caccgccgac aagcagaaga
     1261 acggcatcaa ggtgaacttc aagacccgcc acaacatcga ggacggcagc gtgcagctcg
     1321 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1381 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     1441 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     1501 aagctaagca cttcgtggcc gtcgatcgtt taaagggagg tagtgagtcg accagtggat
     1561 cctggaggct tgctgaaggc tgtatgctgt gtaaacatcc tcgactggaa ggttttggcc
     1621 actgactgac cttccagtag gatgtttaca caggacacaa ggcctgttac tagcactcac
     1681 atggaacaaa tggcccagat ctggccgcac tcgagatatc tagacccagc tttcttgtac
     1741 aaagtggttg atctagaggg cccgcggttc gctgatgggg gaggctaact gaaacacgga
     1801 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     1861 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     1921 gataccccac cgtgacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     1981 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     2041 gccatagcat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
     2101 ggcccgtgtc tcaaaatctc tgatggatct gcgcagctgg ggctctaggg ggtatcccca
     2161 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2221 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2281 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2341 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2401 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2461 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2521 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2581 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     2641 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     2701 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     2761 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     2821 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     2881 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     2941 ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat
     3001 agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc aagcctttgt
     3061 ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc atccccatct
     3121 ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg
     3181 tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg
     3241 ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca
     3301 tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag
     3361 ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct
     3421 ctggttatgt gtgggagggc taagcacttc gtggccgagg agcaggactg acacgtgcta
     3481 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     3541 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     3601 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3661 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3721 tatcatgtct gtataccgtc gctcttccgc tgcttcctcg ctcactgact cgctgcgctc
     3781 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3841 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3901 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3961 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4021 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4081 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4141 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4201 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4261 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4321 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4381 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4441 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4501 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4561 cgacgcgtaa ctcacgttaa gggattttgg tcatgggtgg ctcgacgagg gttatttgcc
     4621 gactaccttg gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg
     4681 cgaggccaag cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg
     4741 ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat
     4801 tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc
     4861 gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag
     4921 atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct
     4981 atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa
     5041 gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg
     5101 ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat
     5161 ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt
     5221 tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag
     5281 gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg
     5341 ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct
     5401 cataatgttt aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctcca
     5461 taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga
     5521 ctgtacccca aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc
     5581 gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag
     5641 cttacgaacc gaacaggctt atgtccactg ggttcgtgcc ttcatccgtt tccacggtgt
     5701 gcgtcacccg gcaaccttgg gtagcagcga agtcgaggca tttctgtcct ggctggtcta
     5761 g
//
