LOCUS       sequence-134514-        7147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Small fragment of split HRP fused to the extracellular terminus of 
            NLG1. CAG promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7147)
  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 7147)
  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: CAG sHRPb-NLG
FEATURES             Location/Qualifiers
     source          1..7147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     regulatory      1726..1735
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2179..2220
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      2682..2691
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4650..4666)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4650..4666)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4663..4685)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4674..4690
                     /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(4698..4704,4705..4722,4723..4728))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4743..4764
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4823..5018
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4869..5004
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5024..5158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5061..5080)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5115..5134
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5226..5243)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5397..5985)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5477..5496)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6156..6947,6948..7016))
                     /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     6779..6798
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7017..7121)
                     /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 catggcactt
     1741 cccagatgca tgtggccaaa ttatgtttgg agagctatga tggcatgtgt ggtccacagg
     1801 ggatccggtg ccccattgac tctctgcttg ttgggatgtt tgctacagac ttttcacgta
     1861 ctctctcaaa agaccggtaa cttgtctgca ctggttgact ttgacctgcg caccccaacc
     1921 atcttcgaca ataagtatta cgtcaatctg gaggaacaga agggcctgat tcagtccgac
     1981 caggagctgt tcagttcccc agatgctacc gacactattc ctcttgtgcg cagcttcgct
     2041 aatagcacac aaaccttttt caatgctttc gtggaagcta tggatcggat gggaaatata
     2101 acgcccctga cgggtacgca gggacagata agacggaact gtagggtcgt gaacagcaac
     2161 tctggctcgg gctcgggcgg caagcccatc cccaaccccc tgctgggcct ggacagcacc
     2221 caattgttgg atgatgtaga cccattggtt actactaact ttggcaagat taggggaatt
     2281 aagaaagaac tcaataatga aattttgggt cctgtcattc agtttcttgg ggttccatat
     2341 gccgctccac caacaggaga acatcgtttc cagcctccag aaccaccatc tccctggtct
     2401 gacatccgga acgccactca gtttgctcct gtatgtcccc agaatatcat tgatggcaga
     2461 ttgcctgaag ttatgcttcc tgtgtggttc actaataact tggatgtggt ttcatcatac
     2521 gtccaagacc agagtgaaga ctgtctatac ttaaacatct atgtcccaac tgaagatggt
     2581 ccccttacaa agaaacacac agatgattta ggtgataatg acggtgctga agatgaagat
     2641 attcgggaca gtgggggtcc caaaccagtg atggtgtaca tccatggcgg ctcttacatg
     2701 gaaggtactg gaaatctgta tgatgggagt gtcttggcaa gctatggcaa tgtgatcgtc
     2761 atcacagtca actatcggct tggggtactt ggcttcttga gcacagggga tcaggctgcc
     2821 aaaggaaact acgggctcct tgacctcatc caggccctaa gatggaccag cgagaacatt
     2881 gggttctttg gtggtgaccc cttgcgaatc actgtgtttg gatcaggcgc tgggggttca
     2941 tgtgtcaacc tgctgacttt atcccattat tctgaaggta accgttggag caattcaacc
     3001 aaaggacttt ttcaacgagc aatagctcag agtggaacag ccctttccag ctgggctgtt
     3061 agtttccagc ctgcaaaata cgctagaatt ctggccacaa aagttggctg caatgtttca
     3121 gatacagtag agttagtaga atgcctgcag aagaagcctt acaaagaact tgttgatcaa
     3181 gatgttcaac cagcccgata ccacatagcc tttggacctg tgatcgatgg tgatgtaata
     3241 ccagatgacc ctcagatact gatggaacaa ggagagttcc tcaactatga tataatgttg
     3301 ggagttaacc aaggggaagg gttgaagttt gtcgaaaata tagtagatag tgatgatggt
     3361 gtatcagcca gtgatttcga ctttgctgtt tctaattttg ttgataattt atatggatat
     3421 cctgaaggca aagatgtttt gagagaaacc attaaattca tgtatactga ctgggctgat
     3481 cgccataatc ctgaaactag aaggaagaca ttgttggctt tgtttacgga ccatcaatgg
     3541 gtagcacctg ctgtggccac agcagacctt cactcgaact ttggctcacc tacatacttc
     3601 tatgcctttt atcatcattg ccaaacagat caagttccag cttgggctga tgcagctcat
     3661 ggggatgagg ttccctatgt gttgggaatc cccatgattg gccctacaga gttatttcct
     3721 tgcaatttct ccaagaatga tgtgatgttg agtgcagtag taatgacata ctggacgaat
     3781 tttgctaaaa ctggtgaccc aaatcaacca gttcctcaag acacaaaatt catccatacc
     3841 aaacccaacc gctttgaaga agtagcatgg accagatatt cccagaaaga ccagctttat
     3901 ctccatattg gattaaaacc gagagttaaa gagcattaca gagccaataa ggtaaatctc
     3961 tggctggagc tggtacctca tctgcataat ctcaatgaca tttctcagta tacctcgaca
     4021 acaactaaag tgccatccac ggacatcact ctcagaccta caaggaagaa ttccacacca
     4081 gtcacatcag cctttcccac tgccaaacag gatgatccca agcaacaacc aagccccttc
     4141 tcggtggatc agagggacta ctccacagag ctaagtgtca ctatcgcagt gggggcctct
     4201 ctgctgtttc tcaacatctt ggcttttgca gccctgtact acaagaagga taagaggaga
     4261 catgatgtcc accggaggtg cagccctcag cgcacgacca ccaacgacct aacccatgct
     4321 ccagaagagg aaattatgtc tctccaaatg aagcacactg acttggatca cgagtgtgag
     4381 tccatccatc cacatgaggt ggttcttcgg accgcctgtc ccccagatta tactctagct
     4441 atgaggaggt cacctgatga tattccacta atgacaccta acaccatcac aatgattccc
     4501 aacactatac cagggattca gcccttacat acattcaaca catttactgg aggacagaat
     4561 aatacactgc cccatcccca cccacacccc cattcacatt caacaaccag ggtatagaga
     4621 tctgcggccg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4681 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4741 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4801 aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa
     4861 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4921 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4981 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc
     5041 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5101 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg
     5161 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5221 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5281 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5341 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5401 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5461 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5521 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5581 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5641 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5701 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5761 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5821 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5881 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5941 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6001 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6061 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6121 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6181 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6241 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6301 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6361 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6421 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6481 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6541 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6601 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6661 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6721 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6781 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6841 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6901 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6961 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7021 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7081 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7141 ccacctg
//
