LOCUS       sequence-202017-        4081 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian Expression of mtagBFP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4081)
  AUTHORS   Hirabayashi Y, Kwon SK, Paek H, Pernice WM, Paul MA, Lee J, Erfani 
            P, Raczkowski A, Petrey DS, Pon LA, Polleux F
  TITLE     ER-mitochondria tethering by PDZD8 regulates Ca(2+) dynamics in 
            mammalian neurons.
  JOURNAL   Science. 2017 Nov 3;358(6363):623-630. doi: 10.1126/science.aan6009.
  PUBMED    29097544
REFERENCE   2  (bases 1 to 4081)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2017-11-3- 3"; volume: "358"; issue: 
            "6363"; pages: "623-630"
COMMENT     Sequence Label: pCAG-AKAP-mtagBFP2-UBC6
FEATURES             Location/Qualifiers
     source          1..4081
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             186..884
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(1074..1093)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1139..1194
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1193..1212)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1585..1601)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1585..1601)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1598..1620)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1609..1625
                     /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(1633..1639,1640..1657,1658..1663))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1678..1699
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1758..1953
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1804..1939
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1959..2093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1996..2015)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2050..2069
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2161..2178)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2332..2920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2412..2431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3091..3882,3883..3951))
                     /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     3714..3733
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3952..4056)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcaaagaatt gctcgagatg gcaatccagt tgcgttcgct cttccccttg gcgttgcccg
       61 gaatgctggc cctccttggc tggtggtggt ttttctctcg taaaaaagac ctggagctga
      121 agctgaggat cctgcagagc accgtgccta gggccaggga ccctcctgtg gccaccggac
      181 cggtcatgag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg
      241 tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc
      301 agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
      361 ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc atccccgact
      421 tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg
      481 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg
      541 tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag aaaacactcg
      601 gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa ggcagaaacg
      661 acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacgccaag accacatata
      721 gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg gactacagac
      781 tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag gtggcagtgg
      841 ccagatactg cgacctccct agcaaactgg ggcacaagct taatagcggc ctgcgcagcc
      901 gcgcgcaggc gagcaacagc cgcgtgatgg tgtacatcgg catcgccatc ttcctgttcg
      961 tgggcctgtt catgaaggat taagcggccg caattcactc ctcaggtgca ggctgcctat
     1021 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt
     1081 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     1141 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     1201 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     1261 gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag
     1321 tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt
     1381 tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc
     1441 ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc
     1501 cctcttctct tatggagatc cctcgacctg caccgtcgac cagctggtcg acggtgcacc
     1561 gtcgaccagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     1621 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     1681 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     1741 gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     1801 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     1861 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     1921 ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata
     1981 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2041 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca
     2101 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     2161 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2221 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2281 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2341 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2401 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2461 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2521 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2581 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2641 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2701 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2761 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2821 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2881 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2941 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3001 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3061 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3121 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3181 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3241 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3301 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3361 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3421 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3481 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3541 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3601 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3661 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3721 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3781 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3841 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3901 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3961 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4021 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4081 t
//
