LOCUS       sequence-325289-        6776 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the pHTomato inserted in the synaptophysion in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6776)
  AUTHORS   Li Y, Tsien RW
  TITLE     pHTomato, a red, genetically encoded indicator that enables 
            multiplex interrogation of synaptic activity.
  JOURNAL   Nat Neurosci. 2012 May 27;15(7):1047-53. doi: 10.1038/nn.3126.
  PUBMED    22634730
REFERENCE   2  (bases 1 to 6776)
  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"; doi: "10.1038/nn"; 
            journalName: "Nat Neurosci"; date: "2012-05-27- 27"; volume: "15"; 
            issue: "7"; pages: "1047-53"
COMMENT     Sequence Label: pCMV-sypHTomato
FEATURES             Location/Qualifiers
     source          1..6776
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     638..657
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      733..742
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1318..2022
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKG
                     EIKMRLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1460..1478)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1728..1747
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1922..1945
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             2479..2505
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2527..2544)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2533..2757
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2889..3477)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2969..2988)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3806..3853
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3878..3897
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4085..4879)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTYGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4196..4215)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4806..4825
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(4978..4997)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        5076..5267
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5224..5244
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(5327..6118,6119..6187))
                     /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     5950..5969
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6188..6292)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6319..6774
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6406..6425)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6616..6637
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccctgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      601 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      661 cccaagcttg gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgg
      721 cttggggata tcgccaccat ggacgtggtg aatcagctgg tggctggggg tcagttccgg
      781 gtggtcaagg agccccttgg cttcgtgaag gtgctgcagt gggtctttgc catcttcgcc
      841 tttgctacgt gcggcagcta caccggagag cttcggctga gcgtggagtg tgccaacaag
      901 acggagagtg ccctcaacat cgaagtcgaa tttgagtacc cattcaggct gcaccaagtg
      961 tactttgatg caccctcctg cgttaaaggg ggcactacca agatcttcct agttggtgac
     1021 tactcctcct cggctgaatt ctttgtcacc gtggctgtgt ttgccttcct ctactccatg
     1081 ggggccctgg ccacctacat cttcctgcag aacaagtacc gagagaacaa caaagggcca
     1141 atgatggact tcctggccac agcagtgttc gctttcatgt ggctagttag ctcatccgcc
     1201 tgggccaaag gcctgtccga tgtgaagatg gccactgacc cagagaacat tatcaaggag
     1261 atgcctatgt gccgccagac agatctagcc accggatcca caggtggagc ctcctccgtg
     1321 agcaagggcg aggagaataa catggccatc atcaaggagt tcatgcgctt caaggtgcgc
     1381 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1441 tacgagggca cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc
     1501 tgggacatcc tgtcccctca gttcacgtac ggctccaagg cctacgtgaa gcaccccgcc
     1561 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     1621 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     1681 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     1741 aagaagacca tgggctggga ggcctccacc gagcggatgt accccgagga cggcgccctg
     1801 aagggcgaga tcaagatgag gctgaagctg aaggacggcg gccactacga cgccgaggtc
     1861 aagaccacct acatggccaa gaagcccgtg cagctgcccg gcgcctacaa gaccgacatc
     1921 aagctggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgcgcc
     1981 gagggccgcc actccaccgg cggcatggac gagctgtaca agtccaccgg cgccagcgga
     2041 ggcacaggtt caggtaccgg tggagctcga atcacatgca aggaactgag ggaccctgtg
     2101 acttcaggac tcaacacctc ggtggtgttt ggcttcctga acctggtgct ctgggttggc
     2161 aacctatggt tcgtgttcaa ggagacaggc tgggccgccc cattcatgcg cgcacctcca
     2221 ggcgccccag aaaagcaacc agctcctggc gatgcctacg gcgatgcggg ctatgggcag
     2281 ggccccggag gctatgggcc ccaggactcc tacgggcctc agggtggtta tcaacccgat
     2341 tacgggcagc cagccagcgg cggtggcggt ggctacgggc ctcagggcga ctatgggcag
     2401 caaggctacg gccaacaggg tgcgcccacc tccttctcca atcagatgtc ggatccgaag
     2461 cttcggtacc aagctgccta cccttacgat gtgcccgatt acgcctgagc ggccgctcga
     2521 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2581 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2641 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2701 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     2761 ctgaggcgga aagaaccagt ggcggtaata cggttatcca cagaatcggg ggataacgca
     2821 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2881 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2941 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3001 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3061 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3121 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3181 tccggtaact atcgtcttga gtccaacccg gtaagacgcg acttatcgcc actggcagca
     3241 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3301 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3361 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3421 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3481 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3541 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3601 agttttaaat caatctaaag tatatatgag taacctgagg ctatggcagg gcctgccgcc
     3661 ccgacgttgg ctgcgagccc tgggccttca cccgaacttg gggggtgggg tggggaaaag
     3721 gaagaaacgc gggcgtattg gccccaatgg ggtctcggtg gggtatcgac agagtgccag
     3781 ccctgggacc gaaccccgcg tttatgaaca aacgacccaa caccgtgcgt tttattctgt
     3841 ctttttattg ccgtcatagc gcgggttcct tccggtattg tctccttccg tgtttcagtt
     3901 agcctccccc tagggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc
     3961 cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa
     4021 tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc
     4081 ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg
     4141 cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat
     4201 cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga
     4261 tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatagg
     4321 tcacgacgag atcctcgccg tcgggcatgc tcgccttgag cctggcgaac agttcggctg
     4381 gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc
     4441 gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat
     4501 caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa
     4561 ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg
     4621 cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata
     4681 gccgcgctgc ctcgtcttgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa
     4741 gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct
     4801 gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca
     4861 atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa
     4921 aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgggcct
     4981 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     5041 gcggagttag gggcggaatg ggcggagtta ggggcgggac tatggttgct gactaattga
     5101 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
     5161 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     5221 cacaccctaa ctgacacaca ttccacagct ggttctttcc gcctcaggac tcttcctttt
     5281 tcaataaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5341 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5401 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5461 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5521 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5581 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5641 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5701 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5761 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5821 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5881 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5941 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6001 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6061 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6121 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6181 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6241 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6301 cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     6361 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     6421 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     6481 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     6541 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     6601 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     6661 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     6721 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttac
//
