LOCUS Exported 5651 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCAG APX W41F dimeric (mito matrix) 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5651 /organism="synthetic DNA construct" /mol_type="other DNA" 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(3202..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" primer_bind 3435..3454 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" 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(4660..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 //