LOCUS       sequence-117519-        4311 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 4311)
  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 4311)
  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 hsa-mir-205
FEATURES             Location/Qualifiers
     source          1..4311
                     /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(1316..1340)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1354..1395
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1754..1771
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1794..1811)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1813..1947
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1837..1856)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1891..1910
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2091..2107)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2091..2107)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2104..2126)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2115..2131
                     /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(2139..2145,2146..2163,2164..2169))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2184..2205
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2322..2339)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2493..3081)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2573..2592)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3252..4043,4044..4112))
                     /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     3875..3894
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4113..4217)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4285..4303
                     /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 gctcatatga gcctccttgg
      901 aggatgtgat cctatgggag gggtaggagt attcaggtcc ttgacatctc ccaaatgtgt
      961 gattccggga tgccaaaggc ctttggccag gtaatgcagt gtctacaggc tgaggttgac
     1021 atgcatcccc accctctgag aaaaagatcc tcagacaatc catgtgcttc tcttgtcctt
     1081 cattccaccg gagtctgtct catacccaac cagatttcag tggagtgaag ttcaggaggc
     1141 atggagctga caaccatgag gcctcggcag ccaccgccac caccgccgcc gccaccaccg
     1201 tagcagcagc agcagcagca gcagcagcag cagcagcagc aagagtaact ctgacttagg
     1261 aatagagaca gccagagaga aatgtgatca atgaaagaga catctggaga agcttaccca
     1321 gctttcttgt acaaagtggt tcgatggttc gaaggtaagc ctatccctaa ccctctcctc
     1381 ggtctcgatt ctacgcgttc tagagcagtg gggtcagctt ctaccggggc ggcgtcccgg
     1441 ggtcttggaa ctgcatgcag actgcctgct tgggaaacat acttctttat atgcccatat
     1501 ggacctgcta agctatggaa tgtaaagaag tatgtatctc aggccgggac ctctctcgcc
     1561 gcactgaggg gcactccaca ccacgggggc cgccgcggcc gacagccagc caggcggctg
     1621 tggtgggggt gaggctgggg cggcaggctg cctgggcgtt ggtggacggt cgtgtccccg
     1681 acagcattgt tctgtcactt tgcctgtgtc agcaggtgga agtgctcacc agacatctag
     1741 aacgcgtacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     1801 tgtgccttct aagatccaga catgataaga tacattgatg agtttggaca aaccacaact
     1861 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     1921 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     1981 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg
     2041 gctgattatg atcagtcgac ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg
     2101 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     2161 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     2221 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     2281 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2341 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2401 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2461 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2521 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2581 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2641 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2701 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2761 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2821 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2881 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     2941 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3001 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3061 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3121 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3181 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3241 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3301 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3361 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3421 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3481 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3541 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3601 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3661 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3721 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3781 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     3841 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     3901 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     3961 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4021 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4081 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4141 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4201 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     4261 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg t
//
