LOCUS       sequence-492864-        8274 bp    DNA     circular SYN 17-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8274)
  AUTHORS   Tsai BP, Wang X, Huang L, Waterman ML
  TITLE     Quantitative profiling of in vivo-assembled RNA-protein complexes 
            using a novel integrated proteomic approach.
  JOURNAL   Mol Cell Proteomics. 2011 Apr;10(4):M110.007385. Epub 2011 Feb 1.
  PUBMED    21285413
REFERENCE   2  (bases 1 to 8274)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell 
            Proteomics."; date: "2011-04"; volume: "10(4)"; pages: "M110.007385.
            Epub 2011 Feb 1"
COMMENT     Sequence Label: MS2-HB
FEATURES             Location/Qualifiers
     source          1..8274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        94..473
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        474..677
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             678..853
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    916..1115
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1316..1732
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1662..1684
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1700..1716
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1769..2072
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2073..2276
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2273..2297
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2420..2806
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2-N55K
                     /note="binds with high affinity to a specific stem-loop 
                     structure in the viral RNA (Lim et al., 1994)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGLY"
     CDS             2420..2806
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2
                     /note="binds to a specific stem-loop structure in the viral
                     RNA (Peabody, 1993)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGLY"
     CDS             3203..3220
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3248..3268
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3518..3535
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    join(3565..4140,4141..4143,4144..4152)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3733..3750)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3960..3979
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4172..4771
                     /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(4172..4191)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4668..4688
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5145..5570
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5673..5695)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5831..5851)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5836..6165
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6016..6151
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6187..6206
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6287..6304)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6458..7046)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6538..6557)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7217..8008,8009..8077))
                     /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     7840..7859
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8078..8182)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8250..8268
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gtcttcaaga attagcttgg ccattgcata cgttgtatcc atatcataat atgtacattt
       61 atattggctc atgtccaaca ttaccgccat gttgacattg attattgact agttattaat
      121 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      181 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      241 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      301 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      361 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
      421 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      481 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      541 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      601 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
      661 tctatataag cagagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcttc
      721 cgatagactg cgtcgcccgg gtacccgtat tcccaataaa gcctcttgct gtttgcatcc
      781 gaatcgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccacgacg
      841 ggggtctttc atttgggggc tcgtccggga tttggagacc cctgcccagg gaccaccgac
      901 ccaccaccgg gaggtaagct ggccagcaac ttatctgtgt ctgtccgatt gtctagtgtc
      961 tatgtttgat gttatgcgcc tgcgtctgta ctagttagct aactagctct gtatctggcg
     1021 gacccgtggt ggaactgacg agttctgaac acccggccgc aaccctggga gacgtcccag
     1081 ggactttggg ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac
     1141 cccgtcagga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
     1201 tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag
     1261 catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca
     1321 gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc
     1381 tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg
     1441 gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca
     1501 ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat
     1561 cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc
     1621 taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc
     1681 gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg ccggaattga
     1741 agatctgggg gatcgatcct ctagagtccg ttacataact tacggtaaat ggcccgcctg
     1801 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     1861 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     1921 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     1981 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     2041 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     2101 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     2161 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     2221 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt
     2281 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac
     2341 accgggaccg atccagcctc cgcggccggg aacggtgcat tggaacggac ctgcaggaat
     2401 tgatccgcgg ccgcgcatgg cttctaactt tactcagttc gttctcgtcg acaatggcgg
     2461 aactggcgac gtgactgtcg ccccaagcaa cttcgctaac ggggtcgctg aatggatcag
     2521 ctctaactcg cgttcacagg cttacaaagt aacctgtagc gttcgtcaga gctctgcgca
     2581 gaagcgcaaa tacaccatca aagtcgaggt gcctaaagtg gcaacccaga ctgttggtgg
     2641 tgtagagctt cctgtagccg catggcgttc gtacttaaat atggaactaa ccattccaat
     2701 tttcgctacg aattccgact gcgagcttat tgttaaggca atgcaaggtc tcctaaaaga
     2761 tggaaacccg attccctcag caatcgcagc aaactccggc ctctacggcg caatggcttc
     2821 taactttact cagttcgttc tcgtcgacaa tggcggaact ggcgacgtga ctgtcgcccc
     2881 aagcaacttc gctaacgggg tcgctgaatg gatcagctct aactcgcgtt cacaggctta
     2941 caaagtaacc tgtagcgttc gtcagagctc tgcgcagaag cgcaaataca ccatcaaagt
     3001 cgaggtacct aaagtggcaa cccagactgt tggtggtgta gagcttcctg tagccgcatg
     3061 gcgttcgtac ttaaatatgg aactaaccat tccaattttc gccacgaatt ccgactgcga
     3121 gcttattgtt aaggcaatgc aaggtctcct aaaagatgga aacccgattc cctcagcaat
     3181 cgcagcaaac tccttaatta accatcatca ccaccatcat gactacgata tacccacaac
     3241 cgccagcgag aatctgtact ttcaaggaga gttagggatg aggggttcag ctggaaaggc
     3301 cggtgaaggt gaaatccctg cccctcttgc tggtaccgtt tctaagatac tggtaaaaga
     3361 aggtgacact gttaaagctg gtcaaacagt tctggtgctg gaggctatga aaatggagac
     3421 agaaattaac gctcctactg acggaaaagt tgaaaaggtg ttagttaagg aaagagatgc
     3481 tgttcaaggt ggtcaaggtc taatcaagat cggcgttcat catcaccacc atcattaatg
     3541 agaattccgc cccccccccc cccccccctc tccctccccc cccccctaac gttactggcc
     3601 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     3661 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     3721 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     3781 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     3841 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     3901 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     3961 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     4021 tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa
     4081 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataag
     4141 cttgccacaa cccacaagga gacgaccttc catgaccgag tacaagccca cggtgcgcct
     4201 cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta
     4261 ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca
     4321 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     4381 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     4441 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     4501 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     4561 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     4621 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     4681 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     4741 ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc cacgacccgc agcgcccgac
     4801 cgaaaggagc gcacgacccc atggctccga ccgaagccac ccggggcggc cccgccgacc
     4861 ccgcacccgc ccccgaggcc caccgactct agtcgagggc tgcagcgctg cagaggccga
     4921 gtgcagaact gctccaaagg gacctcaagg ctttccgagg gacactaggc tgactccatc
     4981 gagccagtgt agagataagc ttatcgatta gtccaatttg ttaaagacag gatatcagtg
     5041 gtccaggctc tagttttgac tcaacaatat caccagctga agcctataga gtacgagcca
     5101 tagataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc
     5161 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa
     5221 ctgagaatag agaagttcag atcaaggtca ggaacagatg gaacagggtc gaccctagag
     5281 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
     5341 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
     5401 aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta
     5461 cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg
     5521 ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg
     5581 tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc
     5641 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     5701 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     5761 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtag
     5821 atccggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     5881 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     5941 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     6001 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     6061 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     6121 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tactggctta
     6181 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     6241 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
     6301 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6361 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6421 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6481 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6541 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6601 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6661 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6721 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6781 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6841 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     6901 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6961 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7021 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7081 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7141 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7201 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7261 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7321 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7381 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7441 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7501 ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt
     7561 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7621 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7681 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7741 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7801 tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca
     7861 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7921 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7981 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8041 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8101 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8161 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     8221 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttc
//
