LOCUS       sequence-438861-        5963 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 5963)
  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 5963)
  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-PRDX1
FEATURES             Location/Qualifiers
     source          1..5963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             67..108
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(188..205)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    194..418
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(551..570)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    702..749
                     /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             757..1779
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             757..1779
                     /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    1909..2030
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1946..1965)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2000..2019
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2079..2095)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2079..2095)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2092..2114)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2103..2119
                     /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(2127..2133,2134..2151,2152..2157))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2172..2193
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2310..2327)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2481..3069)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2561..2580)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3240..4031,4032..4100))
                     /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     3863..3882
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4101..4205)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4280..4299)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4471..4850
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4851..5054
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5005..5025
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    5056..5074
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5077..5095
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     5105..5129
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5229..5252
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5256..5279
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5280..5306
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    5322..5346
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(join(5953..5963,1..14))
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ttgtacaaag tggttgatat ccagcacagt ggcggccgct caagtctaga gggcccgcgg
       61 ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg tacctgatcc
      121 gcggccgcat agataactga tccagtgtgc ctcgagtcta gagggcccgt ttaaacccgc
      181 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
      241 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
      301 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
      361 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
      421 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
      481 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
      541 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
      601 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
      661 gaccccaaaa aacttgatta gggtgatggt tcacgtacct agaagttcct attccgaagt
      721 tcctattctc tagaaagtat aggaacttcc ttggccaaaa agcctgaact caccgcgacg
      781 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg
      841 gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg
      901 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg
      961 gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat
     1021 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
     1081 gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag
     1141 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
     1201 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
     1261 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
     1321 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
     1381 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
     1441 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
     1501 gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt
     1561 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
     1621 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
     1681 gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
     1741 aaccgacgcc ccagcactcg tccgagggca aaggaatagc acgtactacg agatttcgat
     1801 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     1861 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     1921 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     1981 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     2041 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     2101 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2161 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2221 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2281 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2341 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2401 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2461 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2521 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2581 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2641 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2701 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2761 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2821 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2881 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2941 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3001 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3061 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3121 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3181 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3241 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3301 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3361 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3421 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3481 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3541 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3601 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3661 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3721 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3781 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3841 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3901 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3961 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4021 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4081 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4141 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4201 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     4261 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     4321 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     4381 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     4441 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
     4501 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     4561 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     4621 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     4681 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     4741 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     4801 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     4861 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     4921 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     4981 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     5041 atataagcag agctctccct atcagtgata gagatctccc tatcagtgat agagatcgtc
     5101 gacgagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     5161 tccatagaag acaccgggac cgatccagcc tccggactct agcgtttaaa cttaagcttg
     5221 gtaccatgca ccaccaccac caccaccacc acatggacta caaggacgac gatgacaagt
     5281 acccttatga cgtgcccgat tacgctttga accagatatc aacaagtttg tacaaaaaag
     5341 caggctccac catgtcttca ggaaatgcta aaattgggca ccctgccccc aacttcaaag
     5401 ccacagctgt tatgccagat ggtcagttta aagatatcag cctgtctgac tacaaaggaa
     5461 aatatgttgt gttcttcttt taccctcttg acttcacctt tgtgtgcccc acggagatca
     5521 ttgctttcag tgatagggca gaagaattta agaaactcaa ctgccaagtg attggtgctt
     5581 ctgtggattc tcacttctgt catctagcat gggtcaatac acctaagaaa caaggaggac
     5641 tgggacccat gaacattcct ttggtatcag acccgaagcg caccattgct caggattatg
     5701 gggtcttaaa ggctgatgaa ggcatctcgt tcaggggcct ttttatcatt gatgataagg
     5761 gtattcttcg gcagatcact gtaaatgacc tccctgttgg ccgctctgtg gatgagactt
     5821 tgagactagt tcaggccttc cagttcactg acaaacatgg ggaagtgtgc ccagctggct
     5881 ggaaacctgg cagtgatacc atcaagcctg atgtccaaaa gagcaaagaa tatttctcca
     5941 agcagaagta ggacccagct ttc
//
