LOCUS       sequence-134511-        6281 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Large fragment of split HRP fused to the extracellular terminus of 
            neurexin3B. CAG promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6281)
  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 6281)
  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 sHRPa-NRX
FEATURES             Location/Qualifiers
     source          1..6281
                     /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"
     CDS             2497..2520
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3412..3438
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3439..3465
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3784..3800)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3784..3800)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3797..3819)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3808..3824
                     /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(3832..3838,3839..3856,3857..3862))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3877..3898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3957..4152
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4003..4138
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4158..4292
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4195..4214)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4249..4268
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4360..4377)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4531..5119)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4611..4630)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5290..6081,6082..6150))
                     /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     5913..5932
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6151..6255)
                     /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 catgcacctg
     1741 agaatccacg cgagacggag ccctcctcgc cggccggcct ggacgcttgg gatctggttc
     1801 ctgttctggg gatgtatcgt cagctctgta tggagtaccg gtcagcttac tccaactttc
     1861 tacgataact cttgtcctaa cgtgagcaac atcgtgcgag atattatcgt gaacgagttg
     1921 cggtctgatc cgaggatagc cgcttcaatt ttgcggctgc atttccacga ttgttttgta
     1981 aacggctgtg atgcttccat cctcctcgac aataccacct ctttcaggac tgaaaaggat
     2041 gccttcggta acgcgaatag tgctagaggg ttttccgtga tcgatagaat gaaagccgct
     2101 gtggagagcg cctgccccgg gacagttagt tgtgcagacc ttctgaccat agcggcccag
     2161 caatccgtga cgctggccgg tggccccagc tggagggtcc ccctgggaag gagggactct
     2221 ctgcaggctt ttctggactt ggcaaacgca aacctgcctg ctcccttctt cactcttccc
     2281 cagctgaagg attccttcag gaatgtcggc ctgaaccgaa gtagtgattt ggtggctctg
     2341 tccggaggcc acacattcgg caagagtcag tgcaggttca tcatggacag actgtataac
     2401 ttttccaaca ccggcttgcc tgaccctacc ctgaacacaa catacttgca gactctcaga
     2461 ggactgtgtc cgctcaatgg cggctcgggc tcgggcgact acaaggatga cgacgataag
     2521 caattgtctt ctaatgtagc ttcctcctcc tccacctctt cctcgccggg gtctcactct
     2581 cagcacgagc accatttcca tggcagcaag catcactcag tgcctatttc tatctatcgt
     2641 tcccctgttt cccttcgagg aggacacgct ggcgctacgt acatctttgg gaaaagtggt
     2701 gggcttatcc tctacacctg gccagccaat gacaggccca gcacgcggtc tgaccgcctt
     2761 gccgtgggct tcagcaccac tgtgaaggat ggcatcttgg tccgcatcga cagtgctcca
     2821 ggacttggtg acttcctcca gcttcacata gaacagggga aaattggagt tgtcttcaac
     2881 attggcacag ttgacatctc catcaaagag gagagaaccc ctgtaaatga cggcaaatac
     2941 catgtggtac gcttcaccag gaacggcggc aacgccaccc tgcaggtgga caactggcca
     3001 gtgaatgaac attatcctac aggccggcag ttaaccatct tcaacactca ggcgcaaata
     3061 gccattggtg gaaaggacaa aggacgcctc ttccaaggcc aactctctgg gctctattat
     3121 gatggtttga aagtactgaa catggcggct gagaacaacc ccaatattaa aatcaatgga
     3181 agtgttcggc tggttggaga agtcccatca attttgggaa caacacagac gacctccatg
     3241 ccaccagaaa tgtctactac tgtcatggaa accactacta caatggcgac taccacaacc
     3301 cgtaagaatc gctctacagc cagcattcag ccaacatcag atgatcttgt ttcatctgct
     3361 gaatgttcaa gtgatgatga agactttgtt gaatgtgagc cgagtacagc atacccatac
     3421 gatgttccag attacgctta tccttacgac gtacctgact atgctaaccc cacggagccg
     3481 ggaatcagac gggttccggg ggcctcagag gtgatccggg agtcgagcag cacaacaggg
     3541 atggtcgtcg gcattgtggc tgctgccgcc ctctgcatct tgatcctcct gtacgccatg
     3601 tacaagtaca ggaacaggga cgaggggtcc tatcaagtgg acgagacgcg gaactacatc
     3661 agcaactccg cccagagcaa cggcacgctc atgaaggaga agcagcagag ctcgaagagc
     3721 ggccacaaga aacagaaaaa caaggacagg gagtattacg tgtaaaagct tggcgtaatc
     3781 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3841 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3901 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat
     3961 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4021 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4081 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4141 ggcttttgca aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4201 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4261 aactcatcaa tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc
     4321 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4381 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4441 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4501 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4561 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4621 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4681 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     4741 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4801 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4861 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4921 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     4981 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5041 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5101 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5161 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5221 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5281 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5341 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5401 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5461 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5521 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5581 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5641 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5701 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5761 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5821 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5881 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5941 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6001 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6061 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6121 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6181 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6241 aataggggtt ccgcgcacat ttccccgaaa agtgccacct g
//
