LOCUS       sequence-13361-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 2
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..2278)
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     protein_bind    complement(2305..2338)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             2347..3369
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2347..3369
                     /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             3376..3432
                     /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(3739..3758)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             4177..4200
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4240..4284
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             4297..4317
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4333..4356
                     /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 tcgcggtttg aggacaaact cttcgcggtc tttccagtgg
     2281 ggatcgacgg tatcgataag ctttataact tcgtatagca tacattatac gaagttatca
     2341 gccaccatga aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag
     2401 ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc
     2461 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac
     2521 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt
     2581 gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc
     2641 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc
     2701 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg
     2761 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat
     2821 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc
     2881 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat
     2941 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc
     3001 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg
     3061 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga
     3121 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg
     3181 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga
     3241 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc
     3301 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg
     3361 gcaaaggaac gcggagccac gaacttctct ctgttaaagc aagcaggaga tgttgaagaa
     3421 aaccccgggc ccatggtgtc caagggcgag gaactgttca ccggcgtggt gcccatcctg
     3481 gtggagctgg acggcgacgt gaacggccac aagttcagcg tgtccggcga gggcgaaggc
     3541 gacgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     3601 ccctggccca ccctggtgac caccctgacc tacggcgtgc agtgcttcag cagatacccc
     3661 gaccacatga agcagcacga tttcttcaag agcgccatgc ccgagggcta cgtccaggag
     3721 cgcaccatct ttttcaagga cgacggcaac tacaagacca gggccgaggt gaagttcgag
     3781 ggcgacacac tggtgaacag gatcgagctg aagggcatcg acttcaaaga ggacggcaac
     3841 atcctgggcc acaagctgga atacaactac aacagccaca acgtgtacat tatggccgac
     3901 aagcagaaga acggcatcaa ggtcaacttc aagatccggc acaacatcga ggacggcagc
     3961 gtgcagctgg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     4021 cccgacaacc actacctgag cacccagagc gctctcagca aggaccccaa cgagaagcgg
     4081 gaccacatgg tgctgaaaga atttgtgaca gccgccggaa tcaccctggg catggacgag
     4141 ctgtacaaga gcagcggccc cagcggcagc agcagcctgg aagtgctgtt ccagggcccc
     4201 ctgagcagct ctggcccctc tggcagcgcc gccaagttca aagaaacagc tgctgctaaa
     4261 ttcgagcggc agcacatgga cagcggagga ggaggcgaga acctgtactt tcagggaggc
     4321 ggaggcggag gggactataa ggacgacgac gacaagctga ccgacgacct gcccagggcc
     4381 cgcgttcgtc ctcactctct accgcatcgc tgtctgcgag ggccagctgt tcggggtgag
     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
//
