LOCUS       sequence-117510-        4206 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 4206)
  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 4206)
  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-lsy-6
FEATURES             Location/Qualifiers
     source          1..4206
                     /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(1211..1235)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1249..1290
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1649..1666
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1689..1706)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1708..1842
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1732..1751)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1786..1805
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1986..2002)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1986..2002)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1999..2021)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2010..2026
                     /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(2034..2040,2041..2058,2059..2064))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2079..2100
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2217..2234)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2388..2976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2468..2487)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3147..3938,3939..4007))
                     /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     3770..3789
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4008..4112)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4180..4198
                     /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 gctcatatgg caattgagca
      901 aagcatctgc aggtgcctga tattttacgg cttccgccca ttaccgtagc tgaaagagta
      961 taaatagaag attcatcgtt caaatgttta atgttcgact tttcaaataa tgtattccta
     1021 ctaatttctg aaaaataagc catcaaatgc gtctagtatc aaaatcatgt aaaaactgta
     1081 aaacagattt tgtatgagac gcatttcgat gacgggcaca acagagacac cgcccataag
     1141 acggtatctt ttgatggtaa gaggtgtgaa tctataatac gactgtgtga tccatgtgtc
     1201 ggttaagctt acccagcttt cttgtacaaa gtggttcgat ggttcgaagg taagcctatc
     1261 cctaaccctc tcctcggtct cgattctacg cgttctagag cagtggggtc agcttctacc
     1321 ggggcggcgt cccggggtct tggaactgca tgcagactgc ctgcttggga aacatacttc
     1381 tttatatgcc catatggacc tgctaagcta tggaatgtaa agaagtatgt atctcaggcc
     1441 gggacctctc tcgccgcact gaggggcact ccacaccacg ggggccgccg cggccgacag
     1501 ccagccaggc ggctgtggtg ggggtgaggc tggggcggca ggctgcctgg gcgttggtgg
     1561 acggtcgtgt ccccgacagc attgttctgt cactttgcct gtgtcagcag gtggaagtgc
     1621 tcaccagaca tctagaacgc gtaccggtca tcatcaccat caccattgag tttaaacccg
     1681 ctgatcagcc tcgactgtgc cttctaagat ccagacatga taagatacat tgatgagttt
     1741 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     1801 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     1861 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     1921 tacaaatgtg gtatggctga ttatgatcag tcgacctgca ggcatgcaag cttggcgtaa
     1981 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     2041 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     2101 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2161 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2221 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2281 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2341 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2401 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2461 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2521 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2581 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2641 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2701 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2761 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2821 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2881 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2941 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3001 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3061 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3121 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3181 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3241 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3301 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3361 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3421 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3481 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3541 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3601 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3661 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3721 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3781 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3841 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3901 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3961 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4021 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4081 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4141 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     4201 tttcgt
//
