LOCUS       sequence-337239-        5469 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 5469)
  AUTHORS   Rhee HW, Zou P, Udeshi ND, Martell JD, Mootha VK, Carr SA, Ting AY
  TITLE     Proteomic Mapping of Mitochondria in Living Cells via Spatially 
            Restricted Enzymatic Tagging.
  JOURNAL   Science. 2013 Jan 31.
  PUBMED    23371551
REFERENCE   2  (bases 1 to 5469)
  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: "Science. 
            2013 Jan 31."
COMMENT     Sequence Label: pCAG HRP-TM
FEATURES             Location/Qualifiers
     source          1..5469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          614..1630
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1554..1576
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1638..1657
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1723..1732
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1729..1788
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             1792..1818
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2770..2799
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2803..2949
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(2971..2987)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2971..2987)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2984..3006)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2995..3011
                     /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(3019..3025,3026..3043,3044..3049))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3064..3085
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3144..3339
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3190..3325
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3345..3479
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3382..3401)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3436..3455
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3547..3564)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3718..4306)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3798..3817)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4477..5268,5269..5337))
                     /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     5100..5119
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5338..5442)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
      541 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
      601 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc
      661 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
      721 cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct
      781 gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc
      841 ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg
      901 cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg
      961 gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgggggga acaaaggctg
     1021 cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca
     1081 accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct
     1141 ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt
     1201 gccgggcggg gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc
     1261 ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc
     1321 gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag
     1381 gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga
     1441 aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc
     1501 tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg
     1561 cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt
     1621 tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga
     1681 attctgcagt cgacggtacc gcgggcccgg gatccaccgg tcgccaccat ggagacagac
     1741 acactcctgc tatgggtact gctgctctgg gttccaggtt ccactggtga ctatccatat
     1801 gatgttccag attatgctgg ggcccagcag ttaaccccta cattctacga caatagctgt
     1861 cccaacgtgt ccaacatcgt tcgcgacaca atcgtcaacg agctcagatc cgatcccagg
     1921 atcgctgctt caatattacg tctgcacttc catgactgct tcgtgaatgg ttgcgacgct
     1981 agcatattac tggacaacac caccagtttc cgcactgaaa aggatgcatt cgggaacgct
     2041 aacagcgcca ggggctttcc agtgatcgat cgcatgaagg ctgccgttga gtcagcatgc
     2101 ccacgaacag tcagttgtgc agacctgctg actatagctg cgcaacagag cgtgactctt
     2161 gcaggcggac cgtcctggag agtgccgctc ggtcgacgtg actccctaca ggcattccta
     2221 gatctggcca acgccaactt gcctgctcca ttcttcaccc tgccccagct gaaggatagc
     2281 tttagaaacg tgggtctgaa tcgctcgagt gaccttgtgg ctctgtccgg aggacacaca
     2341 tttggaaaga accagtgtag gttcatcatg gataggctct acaatttcag caacactggg
     2401 ttacctgacc ccacgctgaa cactacgtat ctccagacac tgagaggctt gtgcccactg
     2461 aatggcaacc tcagtgcact agtggacttt gatctgcgga ccccaaccat cttcgataac
     2521 aagtactatg tgaatctaga ggagcagaaa ggcctgatac agagtgatca agaactgttt
     2581 agcagtccaa acgccactga caccatccca ctggtgagaa gttttgctaa ctctactcaa
     2641 accttcttta acgccttcgt ggaagccatg gaccgtatgg gtaacattac ccctctgacg
     2701 ggtacccaag gccagattcg tctgaactgc agagtggtca acagcaactc tccgcggctg
     2761 caggtcgacg aacaaaaact catctcagaa gaggatctga atgctgtggg ccaggacacg
     2821 caggaggtca tcgtggtgcc acactccttg ccctttaagg tggtggtgat ctcagccatc
     2881 ctggccctgg tggtgctcac catcatctcc cttatcatcc tcatcatgct ttggcagaag
     2941 aagccacgtt agaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3001 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3061 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3121 aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta
     3181 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3241 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3301 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag
     3361 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3421 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc
     3481 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3541 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3601 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3661 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3721 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3781 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3841 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3901 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3961 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4021 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4081 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4141 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4201 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4261 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4321 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4381 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4441 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4501 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4561 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4621 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4681 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4741 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4801 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4861 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4921 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4981 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5041 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5101 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5161 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5221 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5281 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5341 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5401 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5461 gccacctgg
//
