LOCUS       sequence-13360-e        4725 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 4725)
  AUTHORS   Schnutgen F, Ehrmann F, Poser I, Hubner NC, Hansen J, Floss T, 
            Devries I, Wurst W, Hyman A, Mann M, von Melchner H
  TITLE     Resources for proteomics in mouse embryonic stem cells.
  JOURNAL   Nat Methods. 2011 Feb . 8(2):103-4.
  PUBMED    21278719
REFERENCE   2  (bases 1 to 4725)
  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 
            Methods. 2011 Feb . 8(2):103-4."
COMMENT     Sequence Label: pDTnLAPP2A frame 1
FEATURES             Location/Qualifiers
     source          1..4725
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(18..86,87..878)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPPAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(236..255)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1038..1626
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1527..1546
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1780..1797
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2045..2092
                     /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)."
     misc_feature    join(2174..2239,2240..2277)
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     protein_bind    complement(2304..2337)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             2346..3368
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2346..3368
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3375..3431
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(3738..3757)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             4176..4199
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4239..4283
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             4296..4316
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4332..4355
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    4574..4643
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     protein_bind    complement(4662..4695)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
ORIGIN
        1 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
       61 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
      121 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
      181 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
      241 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
      301 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
      361 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
      421 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
      481 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
      541 agcgtgacac cacgatgcct ccagcaatgg caacaacgtt gcgcaaacta ttaactggcg
      601 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
      661 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
      721 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
      781 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
      841 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
      901 atatacttta gattgatttg cggccgcaaa aggatctagg tgaagatcct ttttgataat
      961 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1021 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1081 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1141 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     1201 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1261 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1321 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1381 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1441 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1501 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1561 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     1621 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     1681 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     1741 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     1801 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     1861 aactatggcc atttaatgta aatacttaag aaaaaaaacc aaattaattt tgatacatgc
     1921 tgcatgtgaa agacccccgc tgacgggtag tcaatcactc agaggagacc ctcccaagga
     1981 acagcgagac cacaagtcgg atgcaactgc aagagggttt attggataca cgggtaccga
     2041 attcgaagtt cctattccga agttcctatt ctctagaaag tataggaact tcggtaacct
     2101 ttgagttctc tcagttgggg gcgtagggtc gccgacatga cacccgcggt ctagacaatt
     2161 gtacgtagat ctgtagggcg cagtagtcca gggtttcctt gatgatgtca tacttatcct
     2221 gtcccttttt tttccacagc tcgcggttga ggacaaactc ttcgcggtct ttccagtggg
     2281 gatcgacggt atcgataagc tttataactt cgtatagcat acattatacg aagttatcag
     2341 ccaccatgaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     2401 tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct
     2461 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     2521 aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg
     2581 acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     2641 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca
     2701 tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
     2761 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
     2821 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
     2881 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
     2941 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
     3001 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
     3061 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat
     3121 cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
     3181 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
     3241 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg
     3301 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
     3361 caaaggaacg cggagccacg aacttctctc tgttaaagca agcaggagat gttgaagaaa
     3421 accccgggcc catggtgtcc aagggcgagg aactgttcac cggcgtggtg cccatcctgg
     3481 tggagctgga cggcgacgtg aacggccaca agttcagcgt gtccggcgag ggcgaaggcg
     3541 acgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     3601 cctggcccac cctggtgacc accctgacct acggcgtgca gtgcttcagc agataccccg
     3661 accacatgaa gcagcacgat ttcttcaaga gcgccatgcc cgagggctac gtccaggagc
     3721 gcaccatctt tttcaaggac gacggcaact acaagaccag ggccgaggtg aagttcgagg
     3781 gcgacacact ggtgaacagg atcgagctga agggcatcga cttcaaagag gacggcaaca
     3841 tcctgggcca caagctggaa tacaactaca acagccacaa cgtgtacatt atggccgaca
     3901 agcagaagaa cggcatcaag gtcaacttca agatccggca caacatcgag gacggcagcg
     3961 tgcagctggc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     4021 ccgacaacca ctacctgagc acccagagcg ctctcagcaa ggaccccaac gagaagcggg
     4081 accacatggt gctgaaagaa tttgtgacag ccgccggaat caccctgggc atggacgagc
     4141 tgtacaagag cagcggcccc agcggcagca gcagcctgga agtgctgttc cagggccccc
     4201 tgagcagctc tggcccctct ggcagcgccg ccaagttcaa agaaacagct gctgctaaat
     4261 tcgagcggca gcacatggac agcggaggag gaggcgagaa cctgtacttt cagggaggcg
     4321 gaggcggagg ggactataag gacgacgacg acaagctgac cgacgacctg cccagggccc
     4381 gcgttcgtcc tcactctcta ccgcatcgct gtctgcgagg gccagctgtt ccggggtgag
     4441 tactccctct caaaagcggg catgacttct gcgctaagat tgtcagtttc caaaaacgag
     4501 gaggatttga tattcacctg gcccgcggtg atgcctttga gggtggccgc ggatatcatg
     4561 cattcgtgac ggtctcgaag ccgcggtgcg ggtgccaggg cgtgcccttg ggctccccgg
     4621 gcgcgtactc cacctcaccc atctggtcca cgctgcataa cgaagttcct atactatttg
     4681 aagaatagga acttcggaat aggaacttca gcaacggatc cggcc
//
