LOCUS       sequence-12081-e        4316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector with human U6 promoter driving control Luciferase 
            miR30-based shRNA..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4316)
  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 4316)
  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_hU6miR-Luc-A
FEATURES             Location/Qualifiers
     source          1..4316
                     /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"
     promoter        426..675
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     426..446
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     598..617
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     ncRNA           730..851
                     /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           879..893
                     /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           921..1047
                     /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(1086..1185)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1297..1316)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1375..1394)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(1968..2501)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2690..2695,2696..2712,2713..2718))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      3375..3963
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3864..3883
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4128..4214
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(4306..4316,1..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 ggatcccccc gagtccaaca cccgtgggaa tcccatgggc accatggccc
      241 ctcgctccaa aaatgctttc gcgtctcgca gacactgctc ggtagtttcg gggatcagcg
      301 tttgagtaag agcccgcgtc tgaaccctcc gcgccgcccc ggcccagtgg aaagacgcgc
      361 aggcaaaacg caccacgtga cggagcgtga ccgcgcgccg agcgcgcgcc aaggtcgggc
      421 aggaagaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
      481 gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
      541 gtagaaagta ataatttctt gggtagtttg cagtttttaa aattatgttt taaaatggac
      601 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
      661 aaaggacgaa acaccgtgct cgcttcggca gcacatatac tagtcgacta gggataacag
      721 ggtaattgtt tgaatgaggc ttcagtactt tacagaatcg ttgcctgcac atcttggaaa
      781 cacttgctgg gattacttct tcaggttaac ccaacagaag gctcgagaag gtatattgct
      841 gttgacagtg agcgcccgcc tgaagtctct gattaatagt gaagccacag atgtattaat
      901 cagagacttc aggcggttgc ctactgcctc ggaattcaag gggctacttt aggagcaatt
      961 atcttgttta ctaaaactga ataccttgct atctctttga tacattttta caaagctgaa
     1021 ttaaaatggt ataaattaaa tcactttttt caattggaag actaatgcgt tttcgagata
     1081 tctagaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     1141 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgcagc
     1201 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     1261 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     1321 aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     1381 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc
     1441 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     1501 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     1561 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat
     1621 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     1681 ccctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     1741 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     1801 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
     1861 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     1921 acgaggggaa attaggccgg gaagccgatc tcggcttgaa cgaattgtta ggtggcggta
     1981 cttgggtcga tatcaaagtg catcacttct tcccgtatgc ccaactttgt atagagagcc
     2041 actgcgggat cgtcaccgta atctgcttgc acgtagatca cataagcacc aagcgcgttg
     2101 gcctcatgct tgaggagatt gatgagcgcg gtggcaatgc cctgcctccg gtgctcgccg
     2161 gagactgcga gatcatagat atagatctca ctacgcggct gctcaaacct gggcagaacg
     2221 taagccgcga gagcgccaac aaccgcttct tggtcgaagg cagcaagcgc gatgaatgtc
     2281 ttactacgga gcaagttccc gaggtaatcg gagtccggct gatgttggga gtaggtggct
     2341 acgtctccga actcacgacc gaaaagatca agagcagccc gcatggattt gacttggtca
     2401 gggccgagcc tacatgtgcg aatgatgccc atacttgagc cacctaactt tgttttaggg
     2461 cgactgccct gctgcgtaac atcgttgctg ctgcgtaaca tcgttgctgc tccataacat
     2521 caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac
     2581 aaaaaaacag tcataacaag ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc
     2641 ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac ttgcattaca gtttacgaac
     2701 cgaacaggct tatgtcaact gggttcgtgc cttcatccgt ttccacggtg tgcgtcaccc
     2761 ggcaaccttg ggcagcagcg aagtcgaggc atttctgtcc tggctggcga acgagcgcaa
     2821 ggtttcggtc tccacgcatc gtcaggcatt ggcggccttg ctgttcttct acggcaaggt
     2881 gctgtgcacg gatctgccct ggcttcagga gatcggaaga cctcggccgt cgcggcgctt
     2941 gccggtggtg ctgaccccgg atgaagtggt tcgcatcctc ggttttctgg aaggcgagca
     3001 tcgtttgttc gcccaggact ctagctatag ttctagtggt tggctactaa taggttgtat
     3061 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     3121 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     3181 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     3241 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     3301 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3361 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3421 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3481 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3541 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3601 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3661 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3721 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3781 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3841 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3901 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3961 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4021 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc tagcatggat
     4081 ctcggggacg tctaactact aagcgagagt agggaactgc caggcatcaa ataaaacgaa
     4141 aggctcagtc ggaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     4201 tgagtaggac aaatccgccg ggagcggatt tgaacgttgt gaagcaacgg cccggagggt
     4261 ggcgggcagg acgcccgcca taaactgcca ggcatcaaac taagcagaag gccatc
//
