LOCUS       sequence-56819-e        6410 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 6410)
  AUTHORS   Nishi K, Nishi A, Nagasawa T, Ui-Tei K
  TITLE     Human TNRC6A is an Argonaute-navigator protein for microRNA-mediated
            gene silencing in the nucleus.
  JOURNAL   RNA. 2013 Jan;19(1):17-35. doi: 10.1261/rna.034769.112. Epub 2012 
            Nov 13.
  PUBMED    23150874
REFERENCE   2  (bases 1 to 6410)
  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: 
            "10.1261/rna.034769.112"; journalName: "RNA"; date: "2013-01"; 
            volume: "19"; issue: "1"; pages: "17-35"
COMMENT     Sequence Label: pSUPER-DsRed
FEATURES             Location/Qualifiers
     source          1..6410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(202..221)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(349..371)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(565..584)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     684..706
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(744..762)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        830..934
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(935..1003,1004..1795)
                     /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     complement(1153..1172)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1966..2554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2455..2474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2708..2725
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2842..2863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2878..2883,2884..2901,2902..2908)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2916..2932
                     /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)."
     primer_bind     2921..2943
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2940..2956
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2940..2956
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(3549..4148)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3632..3652)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4129..4148
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(4173..4672)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4221..4240)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     4614..4637
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     promoter        4683..4897
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     4818..4837
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(4973..4989)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4974..4990)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             complement(5008..5424)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(5024..5040)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(5056..5078)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    5491..5832
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             join(5896..6410,1..2)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 ttcccccctt tttctggaga ctaaataaaa tcttttattt tatctatggc tcgtactcta
       61 taggcttcag ctggtgatat tgttgagtca aaactagagc ctggaccact gatatcctgt
      121 ctttaacaaa ttggactaat cgcctctacg ccggacgcat cgtggccggc atcaccggcg
      181 ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc
      241 gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg
      301 ggggactgtt gggcgccatc tccttgcatg caccattcct tgcgccgtcg ttttacaacg
      361 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
      421 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
      481 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
      541 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
      601 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
      661 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
      721 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
      781 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
      841 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
      901 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
      961 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     1021 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     1081 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     1141 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     1201 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     1261 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     1321 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     1381 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     1441 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     1501 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     1561 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     1621 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     1681 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     1741 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     1801 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     1861 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     1921 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1981 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2041 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2101 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2161 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2221 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2281 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2341 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2401 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2461 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2521 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2581 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2641 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2701 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     2761 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     2821 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     2881 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     2941 aggaaacagc tatgaccatg attacgaatt aattctcccc agctcttctg cgctgtccat
     3001 aatgaactat ttcaggagta ttggagtatg tggattcaaa acagtggcta attgagtcag
     3061 gattacttaa tattcgactc ctaccctcag aatgttgttc tccaacttca agaaactgtt
     3121 acccatgaaa gacccccgct gacgggtagt caatcactca gaggagaccc tcccaaggaa
     3181 cagcgagacc acaagtcgga tgcaactgca agagggttta ttggatacac gggtacccgg
     3241 gcgacgcagt ctatcggagg actggcgcgc cgagtgaggg gttgtgggct cttttattga
     3301 gctcggggag cagaagcgcg cgaacagaag cgagaagcga actgattggt tagttcaaat
     3361 aaggcacagg gtcatttcag gtccttgggg caccctggaa acatctgatg gttctctagc
     3421 ttgccaaacc tacaggtggg gtctttcatt cccccctttt tctggagact aaataaaatc
     3481 ttttatttta tcgatgcatg gggtcgtgcg ctcctttcgg tcgggcgctg cgggtcgtgg
     3541 ggcgggcgtc aggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc
     3601 tcgacgtcgg cggtgacggt gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg
     3661 gaggtctcca ggaaggcggg caccccggcg cgctcggccg cctccactcc ggggagcacg
     3721 acggcgctgc ccagaccctt gccctggtgg tcgggcgaga cgccgacggt ggccaggaac
     3781 cacgcgggct ccttgggccg gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc
     3841 agccgggaac cgctcaactc ggccatgcgc gggccgatct cggcgaacac cgcccccgct
     3901 tcgacgctct ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc
     3961 ttgccgatgt cgagcccgac gcgcgtgagg aagagttctt gcagctcggt gacccgctcg
     4021 atgtggcggt ccggatcgac ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg
     4081 agggtgcgta cggccctggg gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac
     4141 tcggtcatgg taagctagct tgggctgcag gtcgaaaggc ccggagatga ggaagaggag
     4201 aacagcgcgg cagacgtgcg cttttgaagc gtgcagaatg ccgggcctcc ggaggacctt
     4261 cgggcgcccg ccccgcccct gagcccgccc ctgagcccgc ccccggaccc accccttccc
     4321 agcctctgag cccagaaagc gaaggagcaa agctgctatt ggccgctgcc ccaaaggcct
     4381 acccgcttcc attgctcagc ggtgctgtcc atctgcacga gactagtgag acgtgctact
     4441 tccatttgtc acgtcctgca cgacgcgagc tgcggggcgg gggggaactt cctgactagg
     4501 ggaggagtag aaggtggcgc gaaggggcca ccaaagaacg gagccggttg gcgcctaccg
     4561 gtggatgtgg aatgtgtgcg aggccagagg ccacttgtgt agcgccaagt gcccagcggg
     4621 gctgctaaag cgcatgctcc agactgcctt gggaaaagcg cctcccctac ccggtagaat
     4681 tcgaacgctg acgtcatcaa cccgctccaa ggaatcgcgg gcccagtgtc actaggcggg
     4741 aacacccagc gcgcgtgcgc cctggcagga agatggctgt gagggacagg ggagtggcgc
     4801 cctgcaatat ttgcatgtcg ctatgtgttc tgggaaatca ccataaacgt gaaatgtctt
     4861 tggatttggg aatcttataa gttctgtatg agaccacaga tcccgccccg taatgcagaa
     4921 gaagagcttc ctgtcactct tcttctgcat tacggggctt ttttaagctt atcgataccg
     4981 tcgacctcga gagatcgatc taattccggc gcctagagaa ggagtgaggg ctggataaag
     5041 ggaggatcga ggcggggtcg aacgaggagg ttcaaggggg agagacgggg cggatggagg
     5101 aagaggaggc ggaggcttag ggtgtacaaa gggcttgacc cagggagggg ggtcaaaagc
     5161 caaggcttcc caggtcacga tgtaggggac ctggtctggg tgtccatgcg ggccaggtga
     5221 aaagaccttg atcttaacct gggtgatgag gtctcggtta aaggtgccgt ctcgcggcca
     5281 tccgacgtta aaggttggcc attctgcaga gcagaaggta acccaacgtc tcttcttgac
     5341 atctaccgac tggttgtgag cgatccgctc gacatctttc cagtgaccta aggtcaaact
     5401 taagggagtg gtaacagtct ggccctaatt ttcagacaaa tacagaaaca cagtcagaca
     5461 gagacaacac agaacgatgc tgcagcgctg cagcagacaa gacgcgcggc ttcggttcca
     5521 aaccgaaagc aaaaattcag acggaggcgg gaactgtttt aggttctcgt ctcctaccag
     5581 aaccacatat cctgacgggg tcggattcca catcgactcc cttcctcagg tcgggccaca
     5641 aaaacggccc ccaaagtccc tgggacgtct cccagggttg cggccgggtg ttcagaactc
     5701 gtcagttcca ccacgggtcc gccagataca gagctagtta gctaactagt acagacgcag
     5761 gcgcaaacat tagatgccgg cacaaacacg cacaaagaca gagacagaca cgaaacgacc
     5821 gctggccagc ttacctcccg gcgggggggt cggtggtccc tgggcagggg tctccaaatc
     5881 tcggtggaac ctccaaatga aagacccccg aggtgggcag tcaatcaatc tgaggagacc
     5941 ctcccaagga tcagcgagtc cacgattcgg atgcaaacag caagaggctt tattgggaat
     6001 acgggtaccc gggcgacgca gtctatcgga ggactggcgc gccgagtgag gggttgtggg
     6061 ctcttttatt gagctcgggg agcagaagcg cgcgaacaga agcgagaagc gaactgattg
     6121 gttagttcaa ataaggcaca gggtcatttc aggtccttgg ggcaccctgg aaacatctga
     6181 tggttctcta gaaactgctg agggcgggac cgcatctggg gaccatctgt tcttggccct
     6241 gagccggggc aggaactgct taccacagat atcctgtttg gcccatattc tgctgtctct
     6301 ctgttcctaa ccttgatctg aacttctcta ttctcagtta tgtattttcc atgccttgca
     6361 aaatggcgtt acttaagcta gcttgccaaa cctacaggtg gggtctttca
//
