LOCUS       sequence-149914-        8486 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBac transposon vector, sHRPaNRX3beta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8486)
  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 8486)
  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: pXL-CAG-ICeuI-sHRPaNRX3beta
FEATURES             Location/Qualifiers
     source          1..8486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   746..808
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        1072..1375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1378..1653
                     /label=chicken beta-actin promoter
     intron          1654..2662
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2586..2608
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     protein_bind    2729..2753
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3658..3681
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4573..4599
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4600..4626
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(4956..4980)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5071..5090)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5136..5191
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5190..5209)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   5812..5846
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6337..6353)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6337..6353)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6350..6372)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6361..6377
                     /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(6385..6391,6392..6409,6410..6415))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6430..6451
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6568..6585)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6739..7327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6819..6838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7498..8289,8290..8358))
                     /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     8121..8140
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8359..8463)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcctcgtt cattcacgtt tttgaacccg tggaggacgg
      661 gcagactcgc ggtgcaaatg tgttttacag cgtgatggag cagatgaaga tgctcgacac
      721 gctgcagaac acgcagctag attaacccta gaaagataat catattgtga cgtacgttaa
      781 agataatcat gcgtaaaatt gacgcatgtg ttttatcggt ctgtatatcg aggtttattt
      841 attaatttga atagatatta agttttatta tatttacact tacatactaa taataaattc
      901 aacaaacaat ttatttatgt ttatttattt attaaaaaaa aacaaaaact caaaatttct
      961 tctataaagt aacaaaactt ttatcgaatt cctgcagccc gggggatcca ctagttatta
     1021 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     1081 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     1141 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     1201 ctatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     1261 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     1321 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgggtcg
     1381 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
     1441 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggcgc
     1501 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
     1561 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
     1621 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgttgcc ttcgccccgt
     1681 gccccgctcc gcgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
     1741 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga
     1801 cggctcgttt cttttctgtg gctgcgtgaa agccttaaag ggctccggga gggccctttg
     1861 tgcggggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg
     1921 cggcccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc
     1981 cgcgtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg ggggctgcga
     2041 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc
     2101 ggcggtcggg ctgtaacccc cccctgcacc cccctccccg agttgctgag cacggcccgg
     2161 cttcgggtgc ggggctccgt gcggggcgtg gcgcggggct cgccgtgccg ggcggggggt
     2221 ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg
     2281 aggggcgcgg cggccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
     2341 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctggcggag
     2401 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcgggcgaag cggtgcggcg
     2461 ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc
     2521 tccatctcca gcctcggggc tgccgcaggg ggacggctgc cttcgggggg gacggggcag
     2581 ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca
     2641 tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca
     2701 ttttggcaaa gaattcctcg aggatatcac aagtttgtac aaaaaagcag gctccgaatt
     2761 cgcccttcgt aactataacg gtcctaaggt agcgaagcca ccatgttcat gccttcttct
     2821 ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag
     2881 aattccgcca ccatgcacct gagaatccac gcgagacgga gccctcctcg ccggccggcc
     2941 tggacgcttg ggatctggtt cctgttctgg ggatgtatcg tcagctctgt atggagtacc
     3001 ggtcagctta ctccaacttt ctacgataac tcttgtccta acgtgagcaa catcgtgcga
     3061 gatattatcg tgaacgagtt gcggtctgat ccgaggatag ccgcttcaat tttgcggctg
     3121 catttccacg attgttttgt aaacggctgt gatgcttcca tcctcctcga caataccacc
     3181 tctttcagga ctgaaaagga tgccttcggt aacgcgaata gtgctagagg gttttccgtg
     3241 atcgatagaa tgaaagccgc tgtggagagc gcctgccccg ggacagttag ttgtgcagac
     3301 cttctgacca tagcggccca gcaatccgtg acgctggccg gtggccccag ctggagggtc
     3361 cccctgggaa ggagggactc tctgcaggct tttctggact tggcaaacgc aaacctgcct
     3421 gctcccttct tcactcttcc ccagctgaag gattccttca ggaatgtcgg cctgaaccga
     3481 agtagtgatt tggtggctct gtccggaggc cacacattcg gcaagagtca gtgcaggttc
     3541 atcatggaca gactgtataa cttttccaac accggcttgc ctgaccctac cctgaacaca
     3601 acatacttgc agactctcag aggactgtgt ccgctcaatg gcggctcggg ctcgggcgac
     3661 tacaaggatg acgacgataa gcaattgtct tctaatgtag cttcctcctc ctccacctct
     3721 tcctcgccgg ggtctcactc tcagcacgag caccatttcc atggcagcaa gcatcactca
     3781 gtgcctattt ctatctatcg ttcccctgtt tcccttcgag gaggacacgc tggcgctacg
     3841 tacatctttg ggaaaagtgg tgggcttatc ctctacacct ggccagccaa tgacaggccc
     3901 agcacgcggt ctgaccgcct tgccgtgggc ttcagcacca ctgtgaagga tggcatcttg
     3961 gtccgcatcg acagtgctcc aggacttggt gacttcctcc agcttcacat agaacagggg
     4021 aaaattggag ttgtcttcaa cattggcaca gttgacatct ccatcaaaga ggagagaacc
     4081 cctgtaaatg acggcaaata ccatgtggta cgcttcacca ggaacggcgg caacgccacc
     4141 ctgcaggtgg acaactggcc agtgaatgaa cattatccta caggccggca gttaaccatc
     4201 ttcaacactc aggcgcaaat agccattggt ggaaaggaca aaggacgcct cttccaaggc
     4261 caactctctg ggctctatta tgatggtttg aaagtactga acatggcggc tgagaacaac
     4321 cccaatatta aaatcaatgg aagtgttcgg ctggttggag aagtcccatc aattttggga
     4381 acaacacaga cgacctccat gccaccagaa atgtctacta ctgtcatgga aaccactact
     4441 acaatggcga ctaccacaac ccgtaagaat cgctctacag ccagcattca gccaacatca
     4501 gatgatcttg tttcatctgc tgaatgttca agtgatgatg aagactttgt tgaatgtgag
     4561 ccgagtacag catacccata cgatgttcca gattacgctt atccttacga cgtacctgac
     4621 tatgctaacc ccacggagcc gggaatcaga cgggttccgg gggcctcaga ggtgatccgg
     4681 gagtcgagca gcacaacagg gatggtcgtc ggcattgtgg ctgctgccgc cctctgcatc
     4741 ttgatcctcc tgtacgccat gtacaagtac aggaacaggg acgaggggtc ctatcaagtg
     4801 gacgagacgc ggaactacat cagcaactcc gcccagagca acggcacgct catgaaggag
     4861 aagcagcaga gctcgaagag cggccacaag aaacagaaaa acaaggacag ggagtattac
     4921 gtgtaaggcg cgccgggcgg ccaagggcga attcgaccca gctttcttgt acaaagtggt
     4981 gattcgcggc cgcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc
     5041 caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga
     5101 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
     5161 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     5221 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgcca tatgctggct
     5281 gccatgaaca aaggtggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc
     5341 attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg
     5401 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt
     5461 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatga agatccctcg
     5521 acctgcagcc caagcttgat atctataaca agaaaatata tatataataa gttatcacgt
     5581 aagtagaaca tgaaataaca atataattat cgtatgagtt aaatcttaaa agtcacgtaa
     5641 aagataatca tgcgtcattt tgactcacgc ggtcgttata gttcaaaatc agtgacactt
     5701 accgcattga caagcacgcc tcacgggagc tccaagcggc gactgagatg tcctaaatgc
     5761 acagcgacgg attcgcgcta tttagaaaga gagagcaata tttcaagaat gcatgcgtca
     5821 attttacgca gactatcttt ctagggttaa tctagctgca tcaggatcat atcgtcgggt
     5881 cttttttccg gctcagtcat cgcccaagct ggcgctatct gggcatcggg gaggaagaag
     5941 cccgtgcctt ttcccgcgag gttgaagcgg catggaaaga gtttgccgag gatgactgct
     6001 gctgcattga cgttgagcga aaacgcacgt ttaccatgat gattcgggaa ggtgtggcca
     6061 tgcacgcctt taacggtgaa ctgttcgttc aggccacctg ggataccagt tcgtcgcggc
     6121 ttttccggac acagttccgg atggtcagcc cgaagcgcat cagcaacccg aacaataccg
     6181 gcgacagccg gaactgccgt gccggtgtgc agattaatga cagcggtgcg gcgctgggat
     6241 attacgtcag cgaggacggg tatcctggct ggatgccgca gaaatggaca tggatacccc
     6301 gtgagttacc cggcgggcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6361 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6421 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6481 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6541 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6601 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6661 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6721 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6781 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6841 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6901 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6961 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7021 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7081 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7141 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     7201 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7261 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7321 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7381 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7441 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7501 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7561 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7621 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7681 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7741 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7801 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7861 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7921 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7981 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8041 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8101 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8161 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8221 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8281 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8341 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8401 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8461 gcgcacattt ccccgaaaag tgccac
//
