LOCUS       sequence-134538-        7704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Large fragment of split HRP fused to the N-terminus of the yeast 
            mating protein Aga1P. When linearized, this vector incorporates into
            the yeast genome..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7704)
  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 7704)
  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: YIP sHRPa-Aga1P
FEATURES             Location/Qualifiers
     source          1..7704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        269..912
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     806..829
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             1699..1740
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(4202..4219)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4202..4218)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4211..4233)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4427..4446)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4546..4568
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4686..5489)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(5490..5705)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5712..5730)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5798..5902
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5903..5971,5972..6763)
                     /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(6121..6140)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6934..7522
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7423..7442
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7676..7693
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgcctgcag gtcgactcta gaagtttatc attatcaata ctcgccattt caaagaatac
      301 gtaaataatt aatagtagtg attttcctaa ctttatttag tcaaaaaatt agccttttaa
      361 ttctgctgta acccgtacat gcccaaaata gggggcgggt tacacagaat atataacatc
      421 gtaggtgtct gggtgaacag tttattcctg gcatccacta aatataatgg agcccgcttt
      481 ttaagctggc atccagaaaa aaaaagaatc ccagcaccaa aatattgttt tcttcaccaa
      541 ccatcagttc ataggtccat tctcttagcg caactacaga gaacaggggc acaaacaggc
      601 aaaaaacggg cacaacctca atggagtgat gcaacctgcc tggagtaaat gatgacacaa
      661 ggcaattgac ccacgcatgt atctatctca ttttcttaca ccttctatta ccttctgctc
      721 tctctgattt ggaaaaagct gaaaaaaaag gttgaaacca gttccctgaa attattcccc
      781 tacttgacta ataagtatat aaagacggta ggtattgatt gtaattctgt aaatctattt
      841 cttaaacttc ttaaattcta cttttatagt tagtcttttt tttagtttta aaacaccaga
      901 acttagtttc gacggatgga tccagctaaa aaaaccaaaa aatttcagat gacattatct
      961 ttcgctcatt ttacctacct gttcacaata ttgttgggat taactaatat tgccttggca
     1021 tctgatccag aaacgggagg ttctcttaag cagcttactc caactttcta cgataactct
     1081 tgtcctaacg tgagcaacat cgtgcgagat attatcgtga acgagttgcg gtctgatccg
     1141 aggatagccg cttcaatttt gcggctgcat ttccacgatt gttttgtaaa cggctgtgat
     1201 gcttccatcc tcctcgacaa taccacctct ttcaggactg aaaaggatgc cttcggtaac
     1261 gcgaatagtg ctagagggtt ttccgtgatc gatagaatga aagccgctgt ggagagcgcc
     1321 tgccccggga cagttagttg tgcagacctt ctgaccatag cggcccagca atccgtgacg
     1381 ctggccggtg gccccagctg gagggtcccc ctgggaagga gggactctct gcaggctttt
     1441 ctggacttgg caaacgcaaa cctgcctgct cccttcttca ctcttcccca gctgaaggat
     1501 tccttcagga atgtcggcct gaaccgaagt agtgatttgg tggctctgtc cggaggccac
     1561 acattcggca agagtcagtg caggttcatc atggacagac tgtataactt ttccaacacc
     1621 ggcttgcctg accctaccct gaacacaaca tacttgcaga ctctcagagg actgtgtccg
     1681 ctcaatggcg gctcgggcgg caagcccatc cccaaccccc tgctgggcct ggacagcacc
     1741 ggtagcggag gcggagggtc gagcggcgct agcggtggtt ctattctagt gacgataacc
     1801 aagacaaacg atgcaaatgg ggttgttaca actacagttt cacccgcgct agtctccaca
     1861 tccactatcg ttcaagctgg cactacgaca ttgtatacga cttggtgtcc attgacggta
     1921 tccacttcat ctgctgccga aataagtcct tcaatatcgt acgctactac cctatccaga
     1981 tttagtactt tgacattatc tacagaagtc tgctcccatg aggcatgtcc ttcgtcatcg
     2041 acgttgccaa ccaccacctt atctgtgact tccaagttca cttcatatat ttgccctact
     2101 tgtcacacaa ccgctatcag ctcattatcc gaagtaggaa ctacaaccgt ggtatcatcc
     2161 agcgccattg aaccatcaag tgcctctata atctcacctg tcacctctac actttcgagt
     2221 acaacatcgt ccaatccaac tactacctcc ctaagttcga catctacatc tccaagctct
     2281 acatctacat ctccaagctc tacatctacc tcatcaagtt cgacatctac ctcatcaagt
     2341 tcgacatcta cctcatcaag ttcgacatct acatctccaa gttcgacatc cacatcttca
     2401 agtttgacat ccacatcttc aagttctaca tctacatccc aaagttctac atctacctca
     2461 tcaagttcga catctacatc tccaagctct acatctacct catcaagttc aacatctaca
     2521 tctccaagtt ctaaatctac ttctgcaagc tccacttcca cttcttcata ttcaacatct
     2581 acatccccaa gtttgacttc ttcatctcca actttggctt ccacttctcc aagttcaaca
     2641 tctattagct ctacttttac tgattcaact tcatcccttg gctcctctat agcatcttca
     2701 tcaacgtctg tgtcattata cagcccatcc acacctgttt actccgtccc ttcgacttcg
     2761 tcaaatgttg caactccttc tatgacttct tcaactgttg aaacaactgt tagttcacaa
     2821 agttcgtctg aatatatcac caaatcctca atttctacta ctatcccatc attttccatg
     2881 tctacatatt tcaccactgt tagtggagtc actacaatgt atacgacatg gtgtccttat
     2941 agctctgaat ctgagactag cacattaacc agtatgcatg aaacggttac aacagacgct
     3001 acagtctgca ctcacgagtc ttgcatgccc tcgcagacaa caagtttgat tacatcttct
     3061 ataaaaatgt ccactaaaaa cgtcgcaact tctgtaagca cctcaacggt tgaatcctca
     3121 tatgcatgct ccacatgtgc tgaaacgtca cactcgtatt cttccgtgca aacagcttca
     3181 tcaagttctg taacacagca gaccacatcc acaaagagtt gggtaagttc aatgacaact
     3241 tcggatgaag atttcaataa gcacgctacc ggtaagtatc atgtaacatc ttcaggtacc
     3301 tcaaccattt cgactagtgt aagtgaagcc acgagtacat caagcattga ctcagaatct
     3361 caagaacaat catcacactt attatcgaca tcggtccttt catcctcctc cttgtctgct
     3421 acattatcct ctgacagtac tattttgcta ttcagttctg tatcatcact aagtgtcgaa
     3481 cagtcaccag ttaccacact tcaaatttct tcaacatcag agattttaca acccacttct
     3541 tccacagcta ttgctacaat atctgcctct acatcatcac tttccgcaac atctatctct
     3601 acaccatcta cctctgtgga atcgactatt gaatcttcat cattgactcc gacggtatct
     3661 tctattttcc tctcatcatc atctgctccc tcttctctac aaacatctgt taccactaca
     3721 gaagtttcca ctacttcaat ctccatacaa taccaaactt catcaatggt aacaattagc
     3781 caatatatgg gcagtggatc gcaaacgcgt ttgccattag gaaagttggt cttcgccatc
     3841 atggcagttg cttgcaatgt aattttcagt taatagctcg agatctgata acaacagtgt
     3901 agatgtaaca aaatcgactt tgttcccact gtacttttag ctcgtacaaa atacaatata
     3961 cttttcattt ctccgtaaac aacatgtttt cccatgtaat atccttttct atttttcgtt
     4021 ccgttaccaa ctttacacat actttatata gctattcact tctatacact aaaaaactaa
     4081 gacaatttta attttgctgc ctgccatatt tcaatttgtt ataaattcct ataatttatc
     4141 ctattagtag ctaaaaaaag atgaatgtga atcgaatcct aagagaattg agctcgaatt
     4201 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     4261 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     4321 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
     4381 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     4441 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     4501 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     4561 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcggg taataactga
     4621 tataattaaa ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca
     4681 gttttttagt tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa
     4741 cgttcaccct ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata
     4801 atgtcagatc ctgtagagac cacatcatcc acggttctat actgttgacc caatgcgtct
     4861 cccttgtcat ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt
     4921 ccacccatgt ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg
     4981 tcaacagtac ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct
     5041 aacatcaaaa ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta
     5101 acaatacctg ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg
     5161 tatacacccg cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtct
     5221 tcgaagagta aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc
     5281 atggaaaaat cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat
     5341 gcttcaacta actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt
     5401 gtttgctttt cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca
     5461 gcacgttcct tatatgtagc tttcgacatg atttatcttc gtttcctgca tgtttttgtt
     5521 ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt
     5581 atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc
     5641 gaatcaaaaa aatttcaagg aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa
     5701 ttgaaaagct ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     5761 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5821 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5881 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5941 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6001 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6061 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     6121 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     6181 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     6241 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6301 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6361 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6421 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     6481 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6541 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6601 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6661 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6721 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6781 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6841 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6901 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6961 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7021 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7081 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7141 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7201 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7261 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7321 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7381 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7441 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7501 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7561 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     7621 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     7681 gtcagtgagc gaggaagcgg aaga
//
