LOCUS       sequence-117494-        4241 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 4241)
  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 4241)
  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-lin-4
FEATURES             Location/Qualifiers
     source          1..4241
                     /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(1246..1270)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1284..1325
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1684..1701
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1724..1741)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1743..1877
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1767..1786)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1821..1840
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2021..2037)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2021..2037)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2034..2056)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2045..2061
                     /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(2069..2075,2076..2093,2094..2099))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2114..2135
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2252..2269)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2423..3011)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2503..2522)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3182..3973,3974..4042))
                     /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     3805..3824
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4043..4147)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4215..4233
                     /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 taccacaatc
      901 ggtcggactc atcacactta cctttcaaat atctaaacta ttcctgaata taataaatct
      961 tatagttttt agattctaga caatttctag agttttggtt ggtttatgag tttatgcttc
     1021 cggcctgttc cctgagacct caagtgtgag tgtactattg atgcttcaca cctgggctct
     1081 ccgggtacca ggacggtttg agcagatctt tttttctgtt ttcacggggt tttttttcac
     1141 actgggagct agttccaata aaatggtatt cggggaaaag ggtcaaacga ttattttcag
     1201 aagtcgacta caatagagtt ccactaacgc caactgatga agcttaccca gctttcttgt
     1261 acaaagtggt tcgatggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt
     1321 ctacgcgttc tagagcagtg gggtcagctt ctaccggggc ggcgtcccgg ggtcttggaa
     1381 ctgcatgcag actgcctgct tgggaaacat acttctttat atgcccatat ggacctgcta
     1441 agctatggaa tgtaaagaag tatgtatctc aggccgggac ctctctcgcc gcactgaggg
     1501 gcactccaca ccacgggggc cgccgcggcc gacagccagc caggcggctg tggtgggggt
     1561 gaggctgggg cggcaggctg cctgggcgtt ggtggacggt cgtgtccccg acagcattgt
     1621 tctgtcactt tgcctgtgtc agcaggtgga agtgctcacc agacatctag aacgcgtacc
     1681 ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac tgtgccttct
     1741 aagatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     1801 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     1861 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     1921 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg
     1981 atcagtcgac ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     2041 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     2101 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     2161 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     2221 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2281 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2341 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2401 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2461 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2521 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2581 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2641 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2701 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2761 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2821 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     2881 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2941 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3001 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3061 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3121 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3181 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3241 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3301 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3361 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3421 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3481 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3541 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3601 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3661 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3721 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3781 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3841 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3901 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3961 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4021 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4081 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4141 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     4201 cattaaccta taaaaatagg cgtatcacga ggccctttcg t
//
