LOCUS       sequence-12086-e        4259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector with TRE promoter driving mouse G alpha 12 miR30-based 
            shRNA..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4259)
  AUTHORS   Shin KJ, Wall EA, Zavzavadjian JR, Santat LA, Liu J, Hwang JI, 
            Rebres R, Roach T, Seaman W, Simon MI, Fraser ID
  TITLE     A single lentiviral vector platform for microRNA-based conditional 
            RNA interference and coordinated transgene expression.
  JOURNAL   Proc Natl Acad Sci U S A. 2006 Sep 12. 103(37):13759-64.
  PUBMED    16945906
REFERENCE   2  (bases 1 to 4259)
  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: "Proc Natl 
            Acad Sci U S A. 2006 Sep 12. 103(37):13759-64."
COMMENT     Sequence Label: pEN_TmiR_G12-E
FEATURES             Location/Qualifiers
     source          1..4259
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     25..48
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    67..166
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    join(205..223,224..246,247..265,266..288,289..307,308..330,
                     331..349,350..372,373..391,392..414,415..433,434..456,
                     457..475)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        508..546
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..567
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     ncRNA           673..794
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           822..836
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           864..990
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     protein_bind    complement(1029..1128)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1240..1259)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1318..1337)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(1911..2444)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2633..2638,2639..2655,2656..2661))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      3318..3906
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3807..3826
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4071..4157
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(4249..4259,1..17)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctgacggatg gcctttttgc gtttctacaa actcttcctg ttagttagtt acttaagctc
       61 gggccccaaa taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat
      121 aagcaatgct tttttataat gccaactttg tacaaaaaag caggctttaa aggaaccaat
      181 tcagtcgact ggatccttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt
      241 taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg
      301 atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
      361 cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta
      421 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt
      481 gaaagtcgag ctcggtaccc gggtcgaggt aggcgtgtac ggtgggaggc ctatataagc
      541 agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
      601 catagaagac accgggaccg atccagcctc cgcggccccg aactagtcga ctagggataa
      661 cagggtaatt gtttgaatga ggcttcagta ctttacagaa tcgttgcctg cacatcttgg
      721 aaacacttgc tgggattact tcttcaggtt aacccaacag aaggctcgag aaggtatatt
      781 gctgttgaca gtgagcgacg acaccatctt cgacaacatt agtgaagcca cagatgtaat
      841 gttgtcgaag atggtgtcgc tgcctactgc ctcggaattc aaggggctac tttaggagca
      901 attatcttgt ttactaaaac tgaatacctt gctatctctt tgatacattt ttacaaagct
      961 gaattaaaat ggtataaatt aaatcacttt tttcaattgg aagactaatg cgttttcgag
     1021 atatctagac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     1081 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctgc
     1141 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
     1201 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
     1261 ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     1321 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga
     1381 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     1441 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     1501 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag
     1561 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     1621 tgtccctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     1681 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     1741 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
     1801 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     1861 ttgacgaggg gaaattaggc cgggaagccg atctcggctt gaacgaattg ttaggtggcg
     1921 gtacttgggt cgatatcaaa gtgcatcact tcttcccgta tgcccaactt tgtatagaga
     1981 gccactgcgg gatcgtcacc gtaatctgct tgcacgtaga tcacataagc accaagcgcg
     2041 ttggcctcat gcttgaggag attgatgagc gcggtggcaa tgccctgcct ccggtgctcg
     2101 ccggagactg cgagatcata gatatagatc tcactacgcg gctgctcaaa cctgggcaga
     2161 acgtaagccg cgagagcgcc aacaaccgct tcttggtcga aggcagcaag cgcgatgaat
     2221 gtcttactac ggagcaagtt cccgaggtaa tcggagtccg gctgatgttg ggagtaggtg
     2281 gctacgtctc cgaactcacg accgaaaaga tcaagagcag cccgcatgga tttgacttgg
     2341 tcagggccga gcctacatgt gcgaatgatg cccatacttg agccacctaa ctttgtttta
     2401 gggcgactgc cctgctgcgt aacatcgttg ctgctgcgta acatcgttgc tgctccataa
     2461 catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag gcatagactg
     2521 tacaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg
     2581 ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt acagtttacg
     2641 aaccgaacag gcttatgtca actgggttcg tgccttcatc cgtttccacg gtgtgcgtca
     2701 cccggcaacc ttgggcagca gcgaagtcga ggcatttctg tcctggctgg cgaacgagcg
     2761 caaggtttcg gtctccacgc atcgtcaggc attggcggcc ttgctgttct tctacggcaa
     2821 ggtgctgtgc acggatctgc cctggcttca ggagatcgga agacctcggc cgtcgcggcg
     2881 cttgccggtg gtgctgaccc cggatgaagt ggttcgcatc ctcggttttc tggaaggcga
     2941 gcatcgtttg ttcgcccagg actctagcta tagttctagt ggttggctac taataggttg
     3001 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     3061 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     3121 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     3181 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag
     3241 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3301 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3361 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3421 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     3481 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3541 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3601 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3661 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3721 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3781 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3841 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3901 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3961 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgctagcatg
     4021 gatctcgggg acgtctaact actaagcgag agtagggaac tgccaggcat caaataaaac
     4081 gaaaggctca gtcggaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     4141 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgtgaagcaa cggcccggag
     4201 ggtggcgggc aggacgcccg ccataaactg ccaggcatca aactaagcag aaggccatc
//
