LOCUS       sequence-355756-        5013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mRuby3 from the EF1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5013)
  AUTHORS   Vevea JD, Kusick GF, Courtney KC, Chen E, Watanabe S, Chapman ER
  TITLE     Synaptotagmin 7 is targeted to the axonal plasma membrane through 
            gamma-secretase processing to promote synaptic vesicle docking in 
            mouse hippocampal neurons.
  JOURNAL   Elife. 2021 Sep 20;10. pii: 67261. doi: 10.7554/eLife.67261.
  PUBMED    34543184
REFERENCE   2  (bases 1 to 5013)
  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: "Elife."; 
            date: "2021-09-20"; volume: "10. pii: 67261. doi"; pages: " 
            10.7554/eLife.67261"
COMMENT     Sequence Label: pEF-mRuby3
FEATURES             Location/Qualifiers
     source          1..5013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        40..1222
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          271..1213
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1170..1190
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1254..1263
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2039..2058)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2104..2159
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2158..2177)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2520..2536)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2520..2536)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2533..2555)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2544..2560
                     /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(2568..2574,2575..2592,2593..2598))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2613..2634
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2693..2888
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2739..2874
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2894..3028
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2931..2950)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2985..3004
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3096..3113)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3267..3855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3347..3366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4026..4817,4818..4886))
                     /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     4649..4668
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4887..4991)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctgggtcga cattgattat tgactagatc atcgcgtgag gctccggtgc ccgtcagtgg
       61 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc
      121 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
      181 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
      241 tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct
      301 ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacgcc cctggctgca
      361 gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg
      421 cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg
      481 ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc
      541 catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa
      601 tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc
      661 ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat
      721 cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg
      781 tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag
      841 atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga
      901 gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc
      961 atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg
     1021 gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga
     1081 gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc
     1141 cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt
     1201 tcttccattt caggtgtcgt gaggaattct gcagtcgacg gtaccgcggg cccgccacca
     1261 tggtgtctaa gggcgaagag ctgatcaagg aaaatatgcg tatgaaggtg gtcatggaag
     1321 gttcggtcaa cggccaccaa ttcaaatgca caggtgaagg agaaggcaga ccgtacgagg
     1381 gagtgcaaac catgaggatc aaagtcatcg agggaggacc cctgccattt gcctttgaca
     1441 ttcttgccac gtcgttcatg tatggcagcc gtacctttat caagtacccg gccgacatcc
     1501 ctgatttctt taaacagtcc tttcctgagg gttttacttg ggaaagagtt acgagatacg
     1561 aagatggtgg agtcgtcacc gtcacgcagg acaccagcct tgaggatggc gagctcgtct
     1621 acaacgtcaa ggtcagaggg gtaaactttc cctccaatgg tcccgtgatg cagaagaaga
     1681 ccaagggttg ggagcctaat acagagatga tgtatccagc agatggtggt ctgagaggat
     1741 acactgacat cgcactgaaa gttgatggtg gtggccatct gcactgcaac ttcgtgacaa
     1801 cttacaggtc aaaaaagacc gtcgggaaca tcaagatgcc cggtgtccat gccgttgatc
     1861 accgcctgga aaggatcgag gagagtgaca atgaaaccta cgtagtgcaa agagaagtgg
     1921 cagttgccaa atacagcaac cttggtggtg gctaaggccg cactcctcag gtgcaggctg
     1981 cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc
     2041 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     2101 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     2161 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     2221 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
     2281 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     2341 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     2401 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     2461 ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg
     2521 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     2581 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2641 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
     2701 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2761 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2821 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2881 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2941 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3001 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
     3061 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3121 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3181 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3241 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3301 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3361 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3421 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3481 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3541 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3601 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3661 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3721 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3781 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3841 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3901 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3961 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4021 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4081 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4141 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4201 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4261 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4321 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4381 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4441 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4501 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4561 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4621 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4681 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4741 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4801 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4861 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4921 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4981 ggggttccgc gcacatttcc ccgaaaagtg cca
//
