LOCUS       sequence-8084-ex        3566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3566)
  AUTHORS   Oberdoerffer P, Kanellopoulou C, Heissmeyer V, Paeper C, Borowski C,
            Aifantis I, Rao A, Rajewsky K
  TITLE     Efficiency of RNA interference in the mouse hematopoietic system 
            varies between cell types and developmental stages.
  JOURNAL   Mol Cell Biol. 2005 May . 25(10):3896-905.
  PUBMED    15870264
REFERENCE   2  (bases 1 to 3566)
  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: "Mol Cell 
            Biol. 2005 May . 25(10):3896-905."
COMMENT     Sequence Label: pU6-STOP-shA1
FEATURES             Location/Qualifiers
     source          1..3566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..39)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(19..37)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(58..74)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(58..74)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(71..93)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    82..98
                     /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)."
     promoter        complement(join(106..112,113..130,131..136))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    151..172
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(289..306)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(460..1048)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(540..559)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1219..2010,2011..2079))
                     /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     1842..1861
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2080..2184)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2210..2665)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2347..2368)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2559..2578
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2792..2814
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2806..2823
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2807..2823
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2833..2852
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2833..2851
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2875..2891
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2876..2892
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3213..3246
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3279..3299)
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
ORIGIN
        1 ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
       61 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
      121 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
      181 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
      241 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
      301 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
      361 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
      421 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
      481 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
      541 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
      601 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
      661 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
      721 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
      781 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
      841 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
      901 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
      961 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1021 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1081 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1141 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     1201 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     1261 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     1321 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     1381 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     1441 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     1501 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     1561 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     1621 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     1681 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     1741 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     1801 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     1861 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     1921 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     1981 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     2041 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     2101 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     2161 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     2221 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     2281 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     2341 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     2401 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     2461 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     2521 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     2581 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     2641 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa
     2701 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     2761 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     2821 agtgagcgcg cgtaatacga ctcactatag ggcgaattgg agctcggcgc gccctcgagg
     2881 tcgacggtat cgataagctt cccgggaaaa agggaaatgc tctttctcct caaagctttg
     2941 aggagaaaga gcatttcctt cgtataatgt atgctatacg aagttatgga tcccccgggc
     3001 gtggcttggt ggtacacctc tgtcatccca gtaggtaggt agaggcagtc actggagaac
     3061 ctcatcagtg tcaggccaac ctgagctata cagtgagcaa cgagctctgc tgtcaagaaa
     3121 caaaatagaa aaaacgtgag tgagcctcca ataggatgtt agggtgcttg tcagttcctc
     3181 tgaggaacga attcaaaaat cgcaaaaacg taataacttc gtataatgta tgctatacga
     3241 agttatagtc tcaaaacaca caattacttt acagttaggg tgagtttcct tttgtgctgt
     3301 tttttaaaat aataatttag tatttgtatc tcttatagaa atccaagcct atcatgtaaa
     3361 atgtagctag tattaaaaag aacagattat ctgtctttta tcgcacatta agcctctata
     3421 gttactagga aatattatat gcaaattaac cggggcaggg gagtagccga gcttctccca
     3481 caagtctgtg cgagggggcc ggcgcgggcc tagagatggc ggcgtcggat ctagagcggc
     3541 cgcaagcttc gcagatctgg cgcgcc
//
