LOCUS       sequence-51840-e        5651 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 5651)
  AUTHORS   Martell JD, Deerinck TJ, Sancak Y, Poulos TL, Mootha VK, Sosinsky 
            GE, Ellisman MH, Ting AY
  TITLE     Engineered ascorbate peroxidase as a genetically encoded reporter 
            for electron microscopy.
  JOURNAL   Nat Biotechnol. 2012 Oct 21. doi: 10.1038/nbt.2375.
  PUBMED    23086203
REFERENCE   2  (bases 1 to 5651)
  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 
            Biotechnol. 2012 Oct 21. doi: 10.1038/nbt.2375."
COMMENT     Sequence Label: pCAG APX W41F dimeric (mito matrix)
FEATURES             Location/Qualifiers
     source          1..5651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2560..2583
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2675..2694)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2740..2795
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2794..2813)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3154..3170)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3154..3170)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3167..3189)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3178..3194
                     /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(3202..3208,3209..3226,3227..3232))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3247..3268
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3327..3522
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3373..3508
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3528..3662
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3565..3584)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3619..3638
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3730..3747)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3901..4489)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3981..4000)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4660..5451,5452..5520))
                     /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     5283..5302
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5521..5625)
                     /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 tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attccgccac catgctggcc
     1741 acccgcgtgt tcagcctggt gggcaagcgc gccatcagca ccagcgtgtg cgtgcgcgcc
     1801 cacgctagca ggggaaagtc atacccaacc gtgagcccag attaccagaa ggctattgag
     1861 aaggcaaaaa ggaaactgag gggattcatc gcagaaaaga aatgcgcccc actgatcctg
     1921 cgactggctt tccacagcgc aggaacattt gactccaaga ccaaaacagg cggacccttt
     1981 gggactatca agcaccaggc tgaactggca catggggcca acaatggcct ggacattgct
     2041 gtgagactgc tggagccaat caaagaacag ttccccattg tctcctacgc agacttctac
     2101 cagctggccg gcgtggtcgc tgtggagatt accggcggcc ccgaggtccc tttccaccca
     2161 ggaagggagg acaaacctga accccctcca gaggggagac tgccagatgc taccaagggc
     2221 agcgaccatc tgcgggacgt gttcggaaaa gcaatggggc tgtctgacca ggatatcgtg
     2281 gccctgagtg gagggcacac cattggcgcc gctcataagg aacgctcagg cttcgaggga
     2341 ccctggacaa gcaaccctct gatcttcgac aattcctact ttactgaact gctgaccggc
     2401 gagaaggatg gactgctgca gctgccctct gacaaagccc tgctgacaga tagcgtgttc
     2461 aggcctctgg tcgaaaagta tgcagccgac gaggatgtgt tcttcgccga ctatgctgag
     2521 gcacacctga aactgtctga actgggattc gccgaggcag actacaagga tgacgacgat
     2581 aagtaatgag cggccgcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg
     2641 tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg
     2701 gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca
     2761 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
     2821 atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat gccatatgct
     2881 ggctgccatg aacaaaggtg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     2941 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     3001 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     3061 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atgaagatcc
     3121 ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3181 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3241 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3301 gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc
     3361 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3421 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3481 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg
     3541 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3601 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     3661 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3721 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3781 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3841 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3901 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3961 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4021 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4081 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4141 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4201 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4261 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4321 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     4381 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4441 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4501 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4561 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4621 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4681 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4741 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4801 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4861 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4921 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4981 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5041 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5101 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5161 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5221 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5281 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5341 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5401 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5461 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5521 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5581 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5641 agtgccacct g
//
