LOCUS       sequence-117512-        4198 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 4198)
  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 4198)
  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-40
FEATURES             Location/Qualifiers
     source          1..4198
                     /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(1203..1227)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1241..1282
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1641..1658
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1681..1698)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1700..1834
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1724..1743)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1778..1797
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1978..1994)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1978..1994)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1991..2013)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2002..2018
                     /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(2026..2032,2033..2050,2051..2056))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2071..2092
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2209..2226)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2380..2968)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2460..2479)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3139..3930,3931..3999))
                     /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     3762..3781
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4000..4104)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4172..4190
                     /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 gctcatatgc cagctgattt
      901 cgtcttggta ataagctcgt cattgagatt atcaccgggt gtaaatcagc ttggctctgg
      961 tgtctccgaa cctcctgtcc gcacctcagt ggatgtatgc catgatgata agatatcaga
     1021 aatcctatca ccgggtgtac atcagctaag gtgcgggtac aggtgcattt gatatcaagg
     1081 tgagttattt ttaagtgggt cccagagacc ttggtggttt ttctctgcag tgatagatac
     1141 ttctaacaac tcgctatcac cgggtgaaaa atcacctagg tctggagcct cctgctaagc
     1201 ttacccagct ttcttgtaca aagtggttcg atggttcgaa ggtaagccta tccctaaccc
     1261 tctcctcggt ctcgattcta cgcgttctag agcagtgggg tcagcttcta ccggggcggc
     1321 gtcccggggt cttggaactg catgcagact gcctgcttgg gaaacatact tctttatatg
     1381 cccatatgga cctgctaagc tatggaatgt aaagaagtat gtatctcagg ccgggacctc
     1441 tctcgccgca ctgaggggca ctccacacca cgggggccgc cgcggccgac agccagccag
     1501 gcggctgtgg tgggggtgag gctggggcgg caggctgcct gggcgttggt ggacggtcgt
     1561 gtccccgaca gcattgttct gtcactttgc ctgtgtcagc aggtggaagt gctcaccaga
     1621 catctagaac gcgtaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag
     1681 cctcgactgt gccttctaag atccagacat gataagatac attgatgagt ttggacaaac
     1741 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     1801 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     1861 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     1921 tggtatggct gattatgatc agtcgacctg caggcatgca agcttggcgt aatcatggtc
     1981 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     2041 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     2101 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     2161 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2221 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2281 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2341 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2401 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2461 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2521 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2581 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2641 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2701 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2761 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2821 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2881 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2941 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3001 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3061 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3121 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3181 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3241 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3301 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3361 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3421 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3481 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3541 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3601 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3661 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3721 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3781 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3841 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3901 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3961 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4021 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4081 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     4141 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgt
//
