LOCUS       sequence-432934-        7091 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Full-length horseradish peroxidase (HRP) fused to the C-terminus of 
            the yeast mating protein Aga2P. Galactose-inducible expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7091)
  AUTHORS   Martell JD, Yamagata M, Deerinck TJ, Phan S, Kwa CG, Ellisman MH, 
            Sanes JR, Ting AY
  TITLE     A split horseradish peroxidase for the detection of intercellular 
            protein-protein interactions and sensitive visualization of 
            synapses.
  JOURNAL   Nat Biotechnol. 2016 May 30. doi: 10.1038/nbt.3563.
  PUBMED    27240195
REFERENCE   2  (bases 1 to 7091)
  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. 2016 May 30. doi: 10.1038/nbt.3563."
COMMENT     Sequence Label: pCTCON2 Aga2P-HRP
FEATURES             Location/Qualifiers
     source          1..7091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    69..572
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        609..713
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(714..782,783..1574)
                     /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     complement(932..951)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1745..2333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2234..2253
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2487..2504
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2621..2642
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2657..2662,2663..2680,2681..2687)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2695..2711
                     /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)."
     primer_bind     2700..2722
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2719..2735
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2719..2735
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2756..2774
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2804..3456
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    3008..3125
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3420..3443
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             3775..3786
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3787..3813
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3820..3840
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4843..4872
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(5205..5224)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5206..5224)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5231..5248)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5231..5247)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5240..5262)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5388..5843
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5475..5494)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5685..5706
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5946..6620)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(6621..6901)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(6922..6941)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7041..7063
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgga tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat gatggaataa
      121 tttgggaatt tactctgtgt ttatttattt ttatgttttg tatttggatt ttagaaagta
      181 aataaagaag gtagaagagt tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa
      241 tttcaacaaa aagcgtactt tacatatata tttattagac aagaaaagca gattaaatag
      301 atatacattc gattaacgat aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt
      361 ctacacagac aagatgaaac aattcggcat taatacctga gagcaggaag agcaagataa
      421 aaggtagtat ttgttggcga tccccctaga gtcttttaca tcttcggaaa acaaaaacta
      481 ttttttcttt aatttctttt tttactttct atttttaatt tatatattta tattaaaaaa
      541 tttaaattat aattattttt atagcacgtg atgaaaagga cccaggtggc acttttcggg
      601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      721 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      781 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      841 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      901 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
      961 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1021 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1081 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1141 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1201 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1261 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1321 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1381 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1441 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1501 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1561 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1621 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1861 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2161 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2401 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2461 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2521 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     2581 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     2641 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     2701 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctcgaaatta
     2761 accctcacta aagggaacaa aagctggtac caattccttg aattttcaaa aattcttact
     2821 ttttttttgg atggacgcaa agaagtttaa taatcatatt acatggcatt accaccatat
     2881 acatatccat atctaatctt acttatatgt tgtggaaatg taaagagccc cattatctta
     2941 gcctaaaaaa accttctctt tggaactttc agtaatacgc ttaactgctc attgctatat
     3001 tgaagtacgg attagaagcc gccgagcggg tgacagccct ccgaaggaag actctcctcc
     3061 gtgcgtcctc gtcttcaccg gtcgcgttcc tgaaacgcag atgtgcctcg cgccgcactg
     3121 ctccgaacaa taaagattct acaatactag cttttatggt tatgaagagg aaaaattggc
     3181 agtaacctgg ccccacaaac cttcaaatga acgaatcaaa ttaacaacca taggatgata
     3241 atgcgattag ttttttagcc ttatttctgg ggtaattaat cagcgaagcg atgatttttg
     3301 atctattaac agatatataa atgcaaaaac tgcataacca ctttaactaa tactttcaac
     3361 attttcggtt tgtattactt cttattcaaa tgtaataaaa gtatcaacaa aaaattgtta
     3421 atatacctct atactttaac gtcaaggaga aaaaaccccg gatcgaattc cctacttcat
     3481 acattttcaa ttaagatgca gttacttcgc tgtttttcaa tattttctgt tattgcttca
     3541 gttttagcac aggaactgac aactatatgc gagcaaatcc cctcaccaac tttagaatcg
     3601 acgccgtact ctttgtcaac gactactatt ttggccaacg ggaaggcaat gcaaggagtt
     3661 tttgaatatt acaaatcagt aacgtttgtc agtaattgcg gttctcaccc ctcaacaact
     3721 agcaaaggca gccccataaa cacacagtat gtttttaagg acaatagctc gacgattgaa
     3781 ggtagatacc catacgacgt tccagactac gctctgcagg agaacttgta ctttcaaggt
     3841 ctgcaggcta gtggtggagg aggctctggt ggaggcggta gcggaggcgg agggtcggct
     3901 agccagctta ctccaacttt ctacgataac tcttgtccta acgtgagcaa catcgtgcga
     3961 gatactatcg tgaacgagtt gcggtctgat ccgaggatag ccgcttcaat tttgcggctg
     4021 catttccacg attgttttgt aaacggctgt gatgcttcca tcctcctcga caataccacc
     4081 tctttcagga ctgaaaagga tgccttcggt aacgcgaata gtgctagagg gtttcccgtg
     4141 atcgatagaa tgaaagccgc tgtggagagc gcctgccccc ggacagttag ttgtgcagac
     4201 cttctgacca tagcggccca gcaatccgtg acgctggccg gtggccccag ctggagggtc
     4261 cccctgggaa ggagggactc tctgcaggct tttctggact tggcaaacgc aaacctgcct
     4321 gctcccttct tcacccttcc ccagctgaag gattccttca ggaatgtcgg cctgaaccga
     4381 agtagtgatt tggtggctct gtccggaggc cacacattcg gcaagaatca gtgcaggttc
     4441 atcatggaca gactgtataa cttttccaac accggcttgc ctgaccctac cctgaacaca
     4501 acatacttgc agactctcag agggctgtgt ccgctcaatg gcaacttgtc tgcactggtt
     4561 gactttgacc tgcgcacccc aaccatcttc gacaataagt attacgtcaa tctggaggaa
     4621 cagaagggcc tgattcagtc cgaccaggag ctgttcagtt ccccaaatgc taccgacact
     4681 attcctcttg tgcgcagctt cgctaatagc acacaaacct ttttcaatgc tttcgtggaa
     4741 gctatggatc ggatgggaaa tataacgccc ctgacgggta cgcagggaca gataagactg
     4801 aactgtaggg tcgtgaacag caactctggc tcgggcggat ccgaacaaaa gcttatttct
     4861 gaagaggact tgtaatagct cgagatctga taacaacagt gtagatgtaa caaaatcgac
     4921 tttgttccca ctgtactttt agctcgtaca aaatacaata tacttttcat ttctccgtaa
     4981 acaacatgtt ttcccatgta atatcctttt ctatttttcg ttccgttacc aactttacac
     5041 atactttata tagctattca cttctataca ctaaaaaact aagacaattt taattttgct
     5101 gcctgccata tttcaatttg ttataaattc ctataattta tcctattagt agctaaaaaa
     5161 agatgaatgt gaatcgaatc ctaagagaat tgagctccaa ttcgccctat agtgagtcgt
     5221 attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     5281 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     5341 gcaccgatcg cctttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag
     5401 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     5461 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5521 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     5581 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     5641 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5701 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     5761 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5821 caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
     5881 atttcacacc gcatagatcg gcaagtgcac aaacaatact taaataaata ctactcagta
     5941 ataacctatt tcttagcatt tttgacgaaa tttgctattt tgttagagtc ttttacacca
     6001 tttgtctcca cacctccgct tacatcaaca ccaataacgc catttaatct aagcgcatca
     6061 ccaacatttt ctggcgtcag tccaccagct aacataaaat gtaagctttc ggggctctct
     6121 tgccttccaa cccagtcaga aatcgagttc caatccaaaa gttcacctgt cccacctgct
     6181 tctgaatcaa acaagggaat aaacgaatga ggtttctgtg aagctgcact gagtagtatg
     6241 ttgcagtctt ttggaaatac gagtctttta ataactggca aaccgaggaa ctcttggtat
     6301 tcttgccacg actcatctcc atgcagttgg acgatatcaa tgccgtaatc attgaccaga
     6361 gccaaaacat cctccttagg ttgattacga aacacgccaa ccaagtattt cggagtgcct
     6421 gaactatttt tatatgcttt tacaagactt gaaattttcc ttgcaataac cgggtcaatt
     6481 gttctctttc tattgggcac acatataata cccagcaagt cagcatcgga atctagagca
     6541 cattctgcgg cctctgtgct ctgcaagccg caaactttca ccaatggacc agaactacct
     6601 gtgaaattaa taacagacat actccaagct gcctttgtgt gcttaatcac gtatactcac
     6661 gtgctcaata gtcaccaatg ccctccctct tggccctctc cttttctttt ttcgaccgaa
     6721 ttaattctta atcggcaaaa aaagaaaagc tccggatcaa gattgtacgt aaggtgacaa
     6781 gctatttttc aataaagaat atcttccact actgccatct ggcgtcataa ctgcaaagta
     6841 cacatatatt acgatgctgt tctattaaat gcttcctata ttatatatat agtaatgtcg
     6901 tgatctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     6961 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     7021 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     7081 gaaacgcgcg a
//
