LOCUS       sequence-12582-e        2852 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 2852)
  AUTHORS   Trujillo RD, Yue SB, Tang Y, O'Gorman WE, Chen CZ
  TITLE     The potential functions of primary microRNAs in target recognition 
            and repression.
  JOURNAL   EMBO J. 2010 Oct 6;29(19):3272-85. doi: 10.1038/emboj.2010.208. Epub
            2010 Aug 31.
  PUBMED    20808284
REFERENCE   2  (bases 1 to 2852)
  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"; doi: 
            "10.1038/emboj.2010.208"; journalName: "EMBO J"; date: "2010-10-6- 
            6"; volume: "29"; issue: "19"; pages: "3272-85"
COMMENT     Sequence Label: pDPlk-lin-41_LCS-wt
FEATURES             Location/Qualifiers
     source          1..2852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     403..422
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        403..421
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(631..647)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(631..647)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(644..666)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    655..671
                     /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(679..685,686..703,704..709))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    724..745
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(862..879)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1033..1621)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1113..1132)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1792..2583,2584..2652))
                     /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     2415..2434
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2653..2757)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2825..2843
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt attaatacga ctcactatag
      421 ggagaagaga attcctcgag aactcaagta taccttttat acaaccgttc tacactcaac
      481 gcgatgtaaa tatcgcaatc cctttttata caaccattct gcctctgaac cattgaaacc
      541 ttacgcgtgg atcctctaga gtcgacctgc aggcatgcat aagcttttcc tccctttagt
      601 gagggtaatt ctcagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
      661 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
      721 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
      781 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
      841 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
      901 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
      961 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1021 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1081 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1141 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1201 cccttcggga agcgtggcgc tttctcaaag ctcacgctgt aggtatctca gttcggtgta
     1261 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1321 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1381 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1441 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     1501 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1561 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     1621 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     1681 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     1741 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     1801 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     1861 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     1921 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     1981 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     2041 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     2101 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     2161 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     2221 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     2281 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     2341 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     2401 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     2461 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     2521 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     2581 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     2641 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     2701 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     2761 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     2821 taaaaatagg cgtatcacga ggccctttcg tc
//
