LOCUS       sequence-13359-e        4722 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 4722)
  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 4722)
  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 0
FEATURES             Location/Qualifiers
     source          1..4722
                     /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..2276)
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     protein_bind    complement(2303..2336)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             2345..3367
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2345..3367
                     /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             3374..3430
                     /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(3737..3756)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             4175..4198
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4238..4282
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             4295..4315
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4331..4354
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    4571..4640
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     protein_bind    complement(4659..4692)
                     /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 tcgcggtgag gacaaactct tcgcggtctt tccagtgggg
     2281 atcgacggta tcgataagct ttataacttc gtatagcata cattatacga agttatcagc
     2341 caccatgaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
     2401 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
     2461 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
     2521 agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga
     2581 cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     2641 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat
     2701 ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
     2761 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
     2821 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
     2881 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
     2941 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
     3001 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
     3061 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
     3121 gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     3181 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     3241 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga
     3301 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
     3361 aaaggaacgc ggagccacga acttctctct gttaaagcaa gcaggagatg ttgaagaaaa
     3421 ccccgggccc atggtgtcca agggcgagga actgttcacc ggcgtggtgc ccatcctggt
     3481 ggagctggac ggcgacgtga acggccacaa gttcagcgtg tccggcgagg gcgaaggcga
     3541 cgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     3601 ctggcccacc ctggtgacca ccctgaccta cggcgtgcag tgcttcagca gataccccga
     3661 ccacatgaag cagcacgatt tcttcaagag cgccatgccc gagggctacg tccaggagcg
     3721 caccatcttt ttcaaggacg acggcaacta caagaccagg gccgaggtga agttcgaggg
     3781 cgacacactg gtgaacagga tcgagctgaa gggcatcgac ttcaaagagg acggcaacat
     3841 cctgggccac aagctggaat acaactacaa cagccacaac gtgtacatta tggccgacaa
     3901 gcagaagaac ggcatcaagg tcaacttcaa gatccggcac aacatcgagg acggcagcgt
     3961 gcagctggcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     4021 cgacaaccac tacctgagca cccagagcgc tctcagcaag gaccccaacg agaagcggga
     4081 ccacatggtg ctgaaagaat ttgtgacagc cgccggaatc accctgggca tggacgagct
     4141 gtacaagagc agcggcccca gcggcagcag cagcctggaa gtgctgttcc agggccccct
     4201 gagcagctct ggcccctctg gcagcgccgc caagttcaaa gaaacagctg ctgctaaatt
     4261 cgagcggcag cacatggaca gcggaggagg aggcgagaac ctgtactttc agggaggcgg
     4321 aggcggaggg gactataagg acgacgacga caagctgacc gacgacctgc ccagggcccg
     4381 cgttcgtcct cactctctac cgcatcgctg tctgcgaggg ccagctgttg gggtgagtac
     4441 tccctctcaa aagcgggcat gacttctgcg ctaagattgt cagtttccaa aaacgaggag
     4501 gatttgatat tcacctggcc cgcggtgatg cctttgaggg tggccgcgga tatcatgcat
     4561 tcgtgacggt ctcgaagccg cggtgcgggt gccagggcgt gcccttgggc tccccgggcg
     4621 cgtactccac ctcacccatc tggtccacgc tgcataacga agttcctata ctatttgaag
     4681 aataggaact tcggaatagg aacttcagca acggatccgg cc
//
