LOCUS       sequence-308764-        5175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid to express pA-APEX2 for antibody-based 
            biotin proximity labeling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5175)
  TITLE     Bacterial expression plasmid to express pA-APEX2 for antibody-based 
            biotin proximity labeling
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5175)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pD451-pA-APEX2
FEATURES             Location/Qualifiers
     source          1..5175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      547..1135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1036..1055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1289..1306
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1404..1421
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1405..1421
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1885..1904
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1885..1903
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1904..1928
                     /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)."
     RBS             1965..1973
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             1997..2422
                     /codon_start=1
                     /product="immunoglobulin binding domains of Staphylocuccus 
                     aureus protein A"
                     /label=protein A
                     /translation="SLKDDPSQSANLLSEAKKLNESQAPKADNKFNKEQQNAFYEILHL
                     PNLNEEQRNGFIQSLKDDPSQSANLLAEAKKLNDAQAPKADNKFNKEQQNAFYEILHLP
                     NLTEEQRNGFIQSLKDDPSVSKEILAEAKKLNDAQAPK"
     terminator      3214..3261
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3484..3503)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        3570..3661
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3662..4471
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             join(4554..5175,1..461)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4573..4592)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac
       61 aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc
      121 agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata
      181 tctcggtagt gggatacgac gataccgaag atagctcatg ttatatcccg ccgttaacca
      241 ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct
      301 ctcagggcca ggcggtgaag ggcaatcagc tgttgccagt ctcactggtg aaaagaaaaa
      361 ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
      421 agctggcacg acaggtttcc cgactggaaa gcgggcagtg actcatgacc aaaatccctt
      481 aacgtgagtt acgcgcgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
      541 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
      601 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      661 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttagccca
      721 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      781 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      841 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
      901 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
      961 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     1021 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     1081 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     1141 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     1201 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     1261 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaaggcga
     1321 gagtagggaa ctgccaggca tcaaactaag cagaaggccc ctgacggatg gcctttttgc
     1381 gtttctacaa actctttctg tgttgtaaaa cgacggccag tcttaagctc gggccccctg
     1441 ggcggttctg ataacgagta atcgttaatc cgcaaataac gtaaaaaccc gcttcggcgg
     1501 gtttttttat ggggggagtt tagggaaaga gcatttgtca gaatatttaa gggcgcctgt
     1561 cactttgctt gatatatgag aattatttaa ccttataaat gagaaaaaag caacgcactt
     1621 taaataagat acgttgcttt ttcgattgat gaacacctat aattaaacta ttcatctatt
     1681 atttatgatt ttttgtatat acaatatttc tagtttgtta aagagaatta agaaaataaa
     1741 tctcgaaaat aataaaggga aaatcagttt ttgatatcaa aattatacat gtcaacgata
     1801 atacaaaata taatacaaac tataagatgt tatcagtatt tattatgcat ttagaataaa
     1861 ttttgtgtcg cccttccgcg aaattaatac gactcactat aggggaattg tgagcggata
     1921 acaattcccc tctagaaata attttgttta actttttgag accttaagga ggtaaaaaat
     1981 gaccatgatc accccgagcc tgaaagatga cccgagccag agcgcaaacc tgttgtccga
     2041 ggccaaaaag ctgaacgaaa gccaggcgcc gaaagctgac aataagttca ataaagaaca
     2101 acagaatgcc ttttatgaaa ttcttcactt accgaatctg aacgaagaac agcgcaatgg
     2161 ctttatccag agcttgaaag atgatccgag ccagtccgcg aacctgttgg ccgaggccaa
     2221 gaaactgaac gacgctcaag caccgaaagc cgataacaag tttaacaaag aacagcagaa
     2281 tgcgttctac gaaattctgc atttgccgaa cctcaccgaa gaacaacgta atggtttcat
     2341 ccagtctctg aaagacgacc cgtcggtcag caaagaaatc ctggctgagg cgaaaaagtt
     2401 gaacgacgcc caagccccga aagacgatga caaagaattt ggcaaaagct atccaacggt
     2461 cagcgcggat tatcaagatg cagtggagaa agcgaaaaag aaactgcgtg gcttcattgc
     2521 ggagaagcgc tgcgcacctc tgatgctgcg tctggcattc catagcgcgg gtacgttcga
     2581 caagggcacc aagaccggtg gcccgtttgg cacgatcaag cacccggcag agctggcgca
     2641 ctcagcgaac aatggtctgg acattgcagt gcgtctgctg gagccgctga aagctgagtt
     2701 tccgattctg agctacgcgg acttctatca gctggcgggt gtcgttgcgg ttgaagttac
     2761 gggtggccca aaagtgccgt ttcacccggg tcgtgaggat aagcctgagc cgccacctga
     2821 gggtcgcctg ccggatccga ccaagggtag cgatcatctg cgcgacgttt tcggcaaagc
     2881 gatgggcctg acggatcaag acattgtcgc tctgagcggt ggccacacta tcggtgcagc
     2941 gcataaggaa cgtagcggtt tcgagggtcc gtggacctct aatccgctga tcttcgataa
     3001 ctcttacttc accgagttgc tgagcggtga gaaagagggc ctgctgcaac tgccgagcga
     3061 taaggccctg ctgtccgacc cggtgtttcg tccgctggtt gacaaatacg cggcagacga
     3121 agatgcgttt ttcgcggatt acgcagaggc gcaccaaaag ctgagcgagc tgggttttgc
     3181 ggacgcctag tagtagggtt gaggtctcac cccctagcat aaccccttgg ggcctctaaa
     3241 cgggtcttga ggggtttttt gcccctgaga cgcgtcaatc gagttcgtac ctaagggcga
     3301 caccccctaa ttagcccggg cgaaaggccc agtctttcga ctgagccttt cgttttattt
     3361 gatgcctggc agttccctac tctcgcatgg ggagtcccca cactaccatc ggcgctacgg
     3421 cgtttcactt ctgagttcgg catggggtca ggtgggacca ccgcgctact gccgccaggc
     3481 aaacaagggg tgttatgagc catattcagg tataaatggg ctcgcgataa tgttcagaat
     3541 tggttaattg gttgtaacac tgacccctat ttgtttattt ttctaaatac attcaaatat
     3601 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagaa
     3661 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     3721 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     3781 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     3841 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgccacttcc
     3901 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     3961 cggaaaaaca gcgttccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     4021 tgcgctggca gtgttcctgc gccggttgca ctcgattcct gtttgtaatt gtccttttaa
     4081 cagcgatcgc gtatttcgcc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     4141 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     4201 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     4261 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     4321 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     4381 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     4441 atttcatttg atgctcgatg agtttttcta agcggcgcgc catcgaatgg cgcaaaacct
     4501 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatatgaaac
     4561 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
     4621 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
     4681 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
     4741 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     4801 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     4861 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
     4921 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     4981 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     5041 atgaggacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     5101 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
     5161 aatatctcac tcgca
//
