LOCUS       sequence-160479-        7547 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 7547)
  AUTHORS   Wang B, Li S, Qi HH, Chowdhury D, Shi Y, Novina CD
  TITLE     Distinct passenger strand and mRNA cleavage activities of human 
            Argonaute proteins.
  JOURNAL   Nat Struct Mol Biol. 2009 Dec . 16(12):1259-66.
  PUBMED    19946268
REFERENCE   2  (bases 1 to 7547)
  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"; journalName: "Nat Struct 
            Mol Biol. 2009 Dec . 16(12):1259-66."
COMMENT     Sequence Label: pGEX-GST-AGO2
FEATURES             Location/Qualifiers
     source          1..7547
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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)."
     CDS             76..729
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     687..709
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             736..753
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     regulatory      2701..2710
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3415..3437)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3582..3600)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3668..3772
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3773..3841,3842..4633)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3991..4010)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4804..5392
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5293..5312
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5546..5563
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5636..5713
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5714..6796
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5733..5752)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6845..6850,6851..6868,6869..6875)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6883..6899
                     /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     6907..6923
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6907..6923
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             6919..7446
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6939..6956)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6939..6955)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6948..6970)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
      121 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
      181 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
      241 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
      301 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
      361 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
      421 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
      481 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
      541 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
      601 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
      661 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
      721 cctccaaaat cggatctggt tccgcgtgga tccccggaat tcccgggtcg actcgactac
      781 tcgggagccg gccccgcact tgcacctcct gcgccgccgc cccccatcca aggatatgcc
      841 ttcaagcctc cacctagacc cgactttggg acctccggga gaacaatcaa attacaggcc
      901 aatttcttcg aaatggacat ccccaaaatt gacatctatc attatgaatt ggatatcaag
      961 ccagagaagt gcccgaggag agttaacagg gaaatcgtgg aacacatggt ccagcacttt
     1021 aaaacacaga tctttgggga tcggaagccc gtgtttgacg gcaggaagaa tctatacaca
     1081 gccatgcccc ttccgattgg gagggacaag gtggagctgg aggtcacgct gccaggagaa
     1141 ggcaaggatc gcatcttcaa ggtgtccatc aagtgggtgt cctgcgtgag cttgcaggcg
     1201 ttacacgatg cactttcagg gcggctgccc agcgtccctt ttgagacgat ccaggccctg
     1261 gacgtggtca tgaggcactt gccatccatg aggtacaccc ccgtgggccg ctccttcttc
     1321 accgcgtccg aaggctgctc taaccctctt ggcgggggcc gagaagtgtg gtttggcttc
     1381 catcagtccg tccggccttc tctctggaaa atgatgctga atattgatgt gtcagcaaca
     1441 gcgttttaca aggcacagcc agtaatcgag tttgtttgtg aagttttgga ttttaaaagt
     1501 attgaagaac aacaaaaacc tctgacagat tcccaaaggg taaagtttac caaagaaatt
     1561 aaaggtctaa aggtggagat aacgcactgt gggcagatga agaggaagta ccgtgtctgc
     1621 aatgtgaccc ggcggcccgc cagtcaccaa acattcccgc tgcagcagga gagcgggcag
     1681 acggtggagt gcacggtggc ccagtatttc aaggacaggc acaagttggt tctgcgctac
     1741 ccccacctcc catgtttaca agtcggacag gagcagaaac acacctacct tcccctggag
     1801 gtctgtaaca ttgtggcagg acaaagatgt attaaaaaat taacggacaa tcagacctca
     1861 accatgatca gagcgactgc taggtcggcg cccgatcggc aagaagagat tagcaaattg
     1921 atgcgaagtg caagtttcaa cacagatcca tacgtccgtg aatttggaat catggtcaaa
     1981 gatgagatga cagacgtgac tgggcgggtg ctgcagccgc cctccatcct ctacgggggc
     2041 aggaataaag ctattgcgac ccctgtccag ggcgtctggg acatgcggaa caagcagttc
     2101 cacacgggca tcgagatcaa ggtgtgggcc attgcgtgct tcgcccccca gcgccagtgc
     2161 acggaagtcc atctgaagtc cttcacagag cagctcagaa agatctcgag agacgccggc
     2221 atgcccatcc agggccagcc gtgcttctgc aaatacgcgc agggggcgga cagcgtggag
     2281 cccatgttcc ggcacctgaa gaacacgtat gcgggcctgc agctggtggt ggtcatcctg
     2341 cccggcaaga cgcccgtgta cgccgaggtc aagcgcgtgg gagacacggt gctggggatg
     2401 gccacgcagt gcgtgcagat gaagaacgtg cagaggacca cgccacagac cctgtccaac
     2461 ctctgcctga agatcaacgt caagctggga ggcgtgaaca acatcctgct gccccagggc
     2521 aggccgccgg tgttccagca gcccgtcatc tttctgggag cagacgtcac tcaccccccc
     2581 gccggggatg ggaagaagcc ctccattgcc gccgtggtgg gcagcatgga cgcccacccc
     2641 aatcgctact gcgccaccgt gcgcgtgcag cagcaccggc aggagatcat acaagacctg
     2701 gccgccatgg tccgcgagct cctcatccag ttctacaagt ccacgcgctt caagcccacc
     2761 cgcatcatct tctaccgcga cggtgtctct gaaggccagt tccagcaggt tctccaccac
     2821 gagttgctgg ccatccgtga ggcctgtatc aagctagaaa aagactacca gcccgggatc
     2881 accttcatcg tggtgcagaa gaggcaccac acccggctct tctgcactga caagaacgag
     2941 cgggttggga aaagtggaaa cattccagca ggcacgactg tggacacgaa aatcacccac
     3001 cccaccgagt tcgacttcta cctgtgtagt cacgctggca tccaggggac aagcaggcct
     3061 tcgcactatc acgtcctctg ggacgacaat cgtttctcct ctgatgagct gcagattcta
     3121 acctaccagc tgtgtcacac ctacgtgcgc tgcacacgct ccgtgtccat cccagcgcca
     3181 gcatactacg ctcacctggt ggccttccgg gccaggtacc acctggtgga taaggaacat
     3241 gacagtgctg aaggaagcca tacctctggg cagagtaacg ggcgagacca ccaagcactg
     3301 gccaaggcgg tccaggttca ccaagacact ctgcgcacca tgtactttgc ttgagcggcc
     3361 gcatcgtgac tgactgacga tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
     3421 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     3481 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc
     3541 acgtagcgat agcggagtgt ataattcttg aagacgaaag ggcctcgtga tacgcctatt
     3601 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3661 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3721 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3781 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3841 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3901 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3961 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     4021 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4081 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4141 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4201 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4261 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
     4321 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4381 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4441 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4501 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4561 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4621 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4681 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4741 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4801 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4861 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4921 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     4981 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5041 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5101 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5161 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5221 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5281 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5341 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5401 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5461 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     5521 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     5581 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcataa attccgacac
     5641 catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt
     5701 cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc
     5761 ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga
     5821 aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact
     5881 ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc
     5941 gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt
     6001 gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc
     6061 gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt
     6121 ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat
     6181 caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc
     6241 attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct
     6301 gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg
     6361 ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg
     6421 catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc
     6481 cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac
     6541 cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct
     6601 ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa
     6661 tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac
     6721 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     6781 ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcacccc
     6841 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     6901 ttcacacagg aaacagctat gaccatgatt acggattcac tggccgtcgt tttacaacgt
     6961 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     7021 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     7081 ctgaatggcg aatggcgctt tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg
     7141 ctggagtgcg atcttcctga ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac
     7201 ggttacgatg cgcccatcta caccaacgta acctatccca ttacggtcaa tccgccgttt
     7261 gttcccacgg agaatccgac gggttgttac tcgctcacat ttaatgttga tgaaagctgg
     7321 ctacaggaag gccagacgcg aattattttt gatggcgttg gaattagctt atcgactgca
     7381 cggtgcacca atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg gctgtgcagg
     7441 tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg ataatgtttt
     7501 ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctg
//
