LOCUS       sequence-117515-        4196 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 4196)
  AUTHORS   Auyeung VC, Ulitsky I, McGeary SE, Bartel DP
  TITLE     Beyond Secondary Structure: Primary-Sequence Determinants License 
            Pri-miRNA Hairpins for Processing.
  JOURNAL   Cell. 2013 Feb 14;152(4):844-58. doi: 10.1016/j.cell.2013.01.031.
  PUBMED    23415231
REFERENCE   2  (bases 1 to 4196)
  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: 
            "10.1016/j.cell.2013.01"; journalName: "Cell"; date: "2013-02-14- 
            14"; volume: "152"; issue: "4"; pages: "844-58"
COMMENT     Sequence Label: pMT mir-1-1 reporter cel-mir-230
FEATURES             Location/Qualifiers
     source          1..4196
                     /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"
     promoter        407..833
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     protein_bind    859..883
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1207..1231)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1245..1286
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1639..1656
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1679..1696)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1698..1832
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1722..1741)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1776..1795
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1976..1992)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1976..1992)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1989..2011)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2000..2016
                     /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(2024..2030,2031..2048,2049..2054))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2069..2090
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2207..2224)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2378..2966)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2458..2477)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3137..3928,3929..3997))
                     /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     3760..3779
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3998..4102)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4170..4188
                     /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 ccagtgccag tgaattaatt cgttgcagga
      421 caggatgtgg tgcccgatgt gactagctct ttgctgcagg ccgtcctatc ctctggttcc
      481 gataagagac ccagaactcc ggccccccac cgcccaccgc cacccccata catatgtggt
      541 acgcaagtaa gagtgcctgc gcatgcccca tgtgccccac caagagtttt gcatcccata
      601 caagtcccca aagtggagaa ccgaaccaat tcttcgcggg cagaacaaaa gcttctgcac
      661 acgtctccac tcgaatttgg agccggccgg cgtgtgcaaa agaggtgaat cgaacgaaag
      721 acccgtgtgt aaagccgcgt ttccaaaatg tataaaaccg agagcatctg gccaatgtgc
      781 atcagttgtg gtcagcagca aaatcaagtg aatcatctca gtgcaactaa aggggggatc
      841 tagatcgggg tacatcaaac aagtttgtac aaaaaagcag gctcatatgt ccttctttag
      901 cgatcaacca taaaaccccg cctccgtttt gaaacatgaa aaaaagtata aaaatagaat
      961 accacaacga cacttgttaa aatattattc ctcatatcta gcttctgctt gtattttttt
     1021 gtctgatgtt gatgtaatgc cgtcacttgg tcggcgattt aatattatca gaacatagga
     1081 aattgttagt attagttgtg cgaccaggag acggtattcg catatttctt aaatattcat
     1141 tttcataatc gtacaaggta cgtttcgagt aatgtctgca aggttcttgc ccacaatgac
     1201 aagcttaccc agctttcttg tacaaagtgg ttcgatggtt cgaaggtaag cctatcccta
     1261 accctctcct cggtctcgat tctacgcgtt ctagagcagt ggggtcagct tctaccgggg
     1321 cggcgtcccg gggtcttgga actgcatgca gactgcctgc ttgggaaaca tacttcttta
     1381 tatgcccata tggacctgct aagctatgga atgtaaagaa gtatgtatct caggccggga
     1441 cctctctcgc cgcactgagg ggcactccac accacggggg ccgccgcggc cgacagccag
     1501 ccaggcggct gtggtggggg tgaggctggg gcggcaggct gcctgggcgt tggtggacgg
     1561 tcgtgtcccc gccagcattg ttctgtcact ttgcctgtgt cagcaggtgg aagtgctcac
     1621 cagacaacgc gtaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc
     1681 tcgactgtgc cttctaagat ccagacatga taagatacat tgatgagttt ggacaaacca
     1741 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     1801 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     1861 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     1921 gtatggctga ttatgatcag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat
     1981 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2041 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2101 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2161 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2221 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2281 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2341 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2401 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2461 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2521 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2581 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2641 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2701 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2761 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     2821 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2881 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     2941 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3001 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3061 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3121 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3181 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3241 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3301 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3361 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3421 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3481 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3541 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3601 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     3661 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     3721 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     3781 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     3841 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     3901 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3961 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4021 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4081 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     4141 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgt
//
