LOCUS       sequence-12088-e        4641 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector with TRE promoter driving both mouse G alpha 12 and G 
            alpha 13 miR30-based shRNAs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4641)
  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 4641)
  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_G13-L_G12-E
FEATURES             Location/Qualifiers
     source          1..4641
                     /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"
     ncRNA           1055..1176
                     /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           1204..1218
                     /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           1246..1372
                     /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(1411..1510)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1622..1641)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1700..1719)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(2293..2826)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3015..3020,3021..3037,3038..3043))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      3700..4288
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4189..4208
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4453..4539
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(4631..4641,1..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 gtgagcgatg ggtgagtctg taaagtattt agtgaagcca cagatgtaaa
      841 tactttacag actcacccag tgcctactgc ctcggaattc aaggggctac tttaggagca
      901 attatcttgt ttactaaaac tgaatacctt gctatctctt tgatacattt ttacaaagct
      961 gaattaaaat ggtataaatt aaatcacttt tttcaattgg aagactaatg cgttttcgag
     1021 atatctagtc gactagggat aacagggtaa ttgtttgaat gaggcttcag tactttacag
     1081 aatcgttgcc tgcacatctt ggaaacactt gctgggatta cttcttcagg ttaacccaac
     1141 agaaggctcg agaaggtata ttgctgttga cagtgagcga cgacaccatc ttcgacaaca
     1201 ttagtgaagc cacagatgta atgttgtcga agatggtgtc gctgcctact gcctcggaat
     1261 tcaaggggct actttaggag caattatctt gtttactaaa actgaatacc ttgctatctc
     1321 tttgatacat ttttacaaag ctgaattaaa atggtataaa ttaaatcact tttttcaatt
     1381 ggaagactaa tgcgttttcg agatatctag acccagcttt cttgtacaaa gttggcatta
     1441 taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa
     1501 tcattatttg ccatccagct gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     1561 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     1621 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     1681 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     1741 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     1801 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     1861 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     1921 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     1981 aatattgttg atgcgctggc agtgtccctg cgccggttgc attcgattcc tgtttgtaat
     2041 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     2101 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     2161 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     2221 ttctcacttg ataaccttat ttttgacgag gggaaattag gccgggaagc cgatctcggc
     2281 ttgaacgaat tgttaggtgg cggtacttgg gtcgatatca aagtgcatca cttcttcccg
     2341 tatgcccaac tttgtataga gagccactgc gggatcgtca ccgtaatctg cttgcacgta
     2401 gatcacataa gcaccaagcg cgttggcctc atgcttgagg agattgatga gcgcggtggc
     2461 aatgccctgc ctccggtgct cgccggagac tgcgagatca tagatataga tctcactacg
     2521 cggctgctca aacctgggca gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc
     2581 gaaggcagca agcgcgatga atgtcttact acggagcaag ttcccgaggt aatcggagtc
     2641 cggctgatgt tgggagtagg tggctacgtc tccgaactca cgaccgaaaa gatcaagagc
     2701 agcccgcatg gatttgactt ggtcagggcc gagcctacat gtgcgaatga tgcccatact
     2761 tgagccacct aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctgcg
     2821 taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct
     2881 tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg aaaaccgcca
     2941 ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac
     3001 gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt cgtgccttca
     3061 tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc gaggcatttc
     3121 tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag gcattggcgg
     3181 ccttgctgtt cttctacggc aaggtgctgt gcacggatct gccctggctt caggagatcg
     3241 gaagacctcg gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa gtggttcgca
     3301 tcctcggttt tctggaaggc gagcatcgtt tgttcgccca ggactctagc tatagttcta
     3361 gtggttggct actaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     3421 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     3481 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     3541 tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag agcattacgc
     3601 tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt tttcgttcca
     3661 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3721 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3781 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3841 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3901 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3961 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4021 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4081 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4141 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4201 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4261 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4321 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4381 taaccgtatt accgctagca tggatctcgg ggacgtctaa ctactaagcg agagtaggga
     4441 actgccaggc atcaaataaa acgaaaggct cagtcggaag actgggcctt tcgttttatc
     4501 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     4561 gttgtgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat
     4621 caaactaagc agaaggccat c
//
