LOCUS       sequence-352285-        7784 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 7784)
  AUTHORS   Baltz AG, Munschauer M, Schwanhausser B, Vasile A, Murakawa Y, 
            Schueler M, Youngs N, Penfold-Brown D, Drew K, Milek M, Wyler E, 
            Bonneau R, Selbach M, Dieterich C, Landthaler M
  TITLE     The mRNA-Bound Proteome and Its Global Occupancy Profile on 
            Protein-Coding Transcripts.
  JOURNAL   Mol Cell. 2012 Jun 8;46(5):674-90.
  PUBMED    22681889
REFERENCE   2  (bases 1 to 7784)
  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: "Mol Cell.";
            date: "2012-06-8"; volume: "46(5)"; pages: "674-90"
COMMENT     Sequence Label: pFRT/TO/HIS/FLAG/HA-HNRNPU
FEATURES             Location/Qualifiers
     source          1..7784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(2337..2361)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2414..2455
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2535..2552)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2541..2765
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2898..2917)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3049..3096
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3104..4126
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3104..4126
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4256..4377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4293..4312)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4347..4366
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4426..4442)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4426..4442)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4439..4461)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4450..4466
                     /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(4474..4480,4481..4498,4499..4504))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4519..4540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4657..4674)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4828..5416)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4908..4927)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5587..6378,6379..6447))
                     /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     6210..6229
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6448..6552)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6627..6646)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6818..7197
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7198..7401
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7352..7372
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    7403..7421
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7424..7442
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     7452..7476
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             7576..7599
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             7603..7626
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7627..7653
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    7669..7693
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 tctcaaggcc gagctcatgg agcgactcca ggctgcgctg gacgacgagg aggccggggg
       61 ccgccccgcc atggagcccg ggaacggcag cctagacctg ggcggggatt ccgctgggcg
      121 ctcgggagca ggcctcgagc aggaggccgc ggccggcggc gatgaagagg aggaggaaga
      181 ggaagaggag gaggaaggaa tctccgctct ggacggcgac cagatggagc taggagagga
      241 gaacggggcc gcgggggcgg ccgactcggg cccgatggag gaggaggagg ccgcctcgga
      301 agacgagaac ggcgacgatc agggtttcca ggaaggggaa gatgagctcg gggacgaaga
      361 ggaaggcgcg ggcgacgaga acgggcacgg ggagcagcag cctcaaccgc cggcgacgca
      421 gcagcaacag ccccaacagc agcgcggggc cgccaaggag gccgcgggga agagcagcgg
      481 ccccacctcg ctgttcgcgg tgacggtggc gccgcccggg gcgaggcagg gccagcagca
      541 ggcgggaggg gacggcaaaa cagaacagaa aggcggagat aaaaagaggg gtgttaaaag
      601 accacgagaa gatcatggcc gtggatattt tgagtacatt gaagagaaca agtatagcag
      661 agccaaatct cctcagccac ctgttgaaga agaagatgaa cacttcgatg acacagtggt
      721 ttgtcttgat acttataatt gtgatctaca ttttaaaata tcaagagatc gtctcagtgc
      781 ttcttccctt acaatggaga gttttgcttt tctttgggct ggaggaagag catcctatgg
      841 tgtgtcaaaa ggcaaagtgt gttttgagat gaaggttaca gagaagatcc cagtaaggca
      901 tttatataca aaagatattg acatacatga agttcgtatt ggctggtcac taactacaag
      961 tggaatgtta cttggtgaag aagaattttc ttatgggtat tctctaaaag gaataaaaac
     1021 atgcaactgt gagactgaag attatggaga aaagtttgat gaaaatgatg tgattacatg
     1081 ttttgctaac tttgaaagtg atgaagtaga actctcgtat gctaagaatg gacaagatct
     1141 tggcgttgcc ttcaaaatca gtaaggaagt tcttgctgga cggccactgt tcccgcatgt
     1201 tctctgccac aactgtgcag ttgaatttaa ttttggtcag aaggaaaagc catattttcc
     1261 aatacctgaa gagtatactt tcatccagaa cgtcccctta gaggatcgag ttagaggacc
     1321 aaaggggcct gaagagaaga aagattgtga agttgtgatg atgattggct tgccaggagc
     1381 tggaaaaact acctgggtta ctaaacatgc agcagaaaat ccagggaaat ataacattct
     1441 tggcacaaat actattatgg ataagatgat ggtggcaggt tttaagaagc aaatggcaga
     1501 tactggaaaa ctgaacacac tgttgcagag agccccccag tgtcttggga aatttattga
     1561 gattgctgcc cgaaagaagc gaaattttat tctggatcag acaaatgtgt ctgctgctgc
     1621 ccagaggaga aaaatgtgcc tgtttgcagg cttccagcga aaagctgttg tagtttgccc
     1681 aaaagatgaa gactataagc aaagaacaca gaagaaagca gaagtagagg ggaaagacct
     1741 accagaacat gcggtcctca aaatgaaagg aaactttacc ctcccagagg tagctgagtg
     1801 ctttgatgaa ataacctatg ttgaacttca gaaggaagaa gcccaaaaac tcttggagca
     1861 atataaggaa gaaagcaaaa aggctcttcc accagaaaag aaacagaaca ctggctcaaa
     1921 gaaaagcaat aaaaataaga gtggcaagaa ccagtttaac agaggtggtg gccatagagg
     1981 acgtggagga ttcaatatgc gtggtggaaa tttcagagga ggagcccctg ggaatcgtgg
     2041 cggatataat aggaggggca acatgccaca gagaggtggt ggcggtggag gaagtggtgg
     2101 aatcggctat ccataccctc gtgcccctgt ttttcctggc cgtggtagtt actcaaacag
     2161 agggaactac aacagaggtg gaatgcccaa cagagggaac tacaaccaga acttcagagg
     2221 acgaggaaac aatcgtggct acaaaaatca atctcagggc tacaaccagt ggcagcaggg
     2281 tcaattctgg ggtcagaagc catggagtca gcattatcac caaggatatt attaggaccc
     2341 agctttcttg tacaaagtgg ttgatatcca gcacagtggc ggccgctcaa gtctagaggg
     2401 cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac
     2461 ctgatccgcg gccgcataga taactgatcc agtgtgcctc gagtctagag ggcccgttta
     2521 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2581 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2641 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2701 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     2761 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     2821 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2881 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2941 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3001 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtacctaga agttcctatt
     3061 ccgaagttcc tattctctag aaagtatagg aacttccttg gccaaaaagc ctgaactcac
     3121 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     3181 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     3241 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     3301 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct
     3361 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     3421 actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct
     3481 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     3541 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     3601 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     3661 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     3721 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     3781 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     3841 ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct
     3901 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     3961 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     4021 acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga
     4081 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagcacg tactacgaga
     4141 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4201 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4261 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4321 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4381 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     4441 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4501 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4561 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4621 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4681 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4741 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4801 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4861 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4921 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4981 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5041 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5101 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5161 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5221 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5281 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5341 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5401 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5461 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5521 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5581 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5641 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5701 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5761 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5821 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5881 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5941 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6001 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6061 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6121 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6181 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6241 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6301 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6361 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6421 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6481 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6541 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     6601 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     6661 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     6721 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
     6781 gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat
     6841 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     6901 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     6961 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     7021 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     7081 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     7141 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     7201 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     7261 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     7321 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     7381 gaggtctata taagcagagc tctccctatc agtgatagag atctccctat cagtgataga
     7441 gatcgtcgac gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt
     7501 tttgacctcc atagaagaca ccgggaccga tccagcctcc ggactctagc gtttaaactt
     7561 aagcttggta ccatgcacca ccaccaccac caccaccaca tggactacaa ggacgacgat
     7621 gacaagtacc cttatgacgt gcccgattac gctttgaacc agatatcaac aagtttgtac
     7681 aaaaaagcag gctccaccat gagttcctcg cctgttaatg taaaaaagct gaaggtgtcg
     7741 gagctgaaag aggagctcaa gaagcgacgc ctttctgaca aggg
//
