LOCUS       sequence-176911-        7779 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Beta1b mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7779)
  AUTHORS   Page KM, Rothwell SW, Dolphin AC
  TITLE     The CaVbeta Subunit Protects the I-II Loop of the Voltage-gated 
            Calcium Channel CaV2.2 from Proteasomal Degradation but Not 
            Oligoubiquitination.
  JOURNAL   J Biol Chem. 2016 Sep 23;291(39):20402-16. doi: 
            10.1074/jbc.M116.737270. Epub 2016 Aug 3.
  PUBMED    27489103
REFERENCE   2  (bases 1 to 7779)
  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: 
            "10.1074/jbc.M116.737270"; journalName: "J Biol Chem"; date: 
            "2016-09-23- 23"; volume: "291"; issue: "39"; pages: "20402-16"
COMMENT     Sequence Label: pMT2 Beta1b mCherry
FEATURES             Location/Qualifiers
     source          1..7779
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1798..2505
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1940..1958)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2208..2227
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2620..3183
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    3370..3504
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3407..3426)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3461..3480
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4177..4194)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4348..4936)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4428..4447)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5107..5898,5899..5967))
                     /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     5730..5749
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5968..6072)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6140..6158
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6196..6218)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6318..6337
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6531..6553
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6545..6562
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6546..6562
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6572..6901)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6586..6721
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6886..6906
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     intron          7370..7599
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     7582..7601
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 atggtccaga agagcggcat gtcccggggc ccttacccac cttcccaaga gatccctatg
       61 gaggtcttcg accccagccc acagggcaaa tacagcaaga ggaaagggcg gttcaaaagg
      121 tcagatggga gcacctcctc agataccaca tccaacagct ttgtccgtca gggctcagca
      181 gagtcctaca cgagccggcc gtcagactcc gacgtgtccc tggaggagga ccgggaagcc
      241 ttaaggaagg aggcagagcg ccaggcctta gcccagctcg agaaagccaa gaccaaacca
      301 gtggcctttg ctgttcggac aaatgttggc tacaatccgt ctccaggaga tgaggtgcca
      361 gtgcagggag tggccatcac ctttgagccc aaggacttcc tgcacatcaa ggagaaatac
      421 aataatgact ggtggatcgg gaggctggtg aaggaaggct gtgaggttgg tttcatcccc
      481 agccctgtca aactggacag ccttcgcctg ctgcaggaac agacgctgcg ccaaaaccgc
      541 ctcagctcca gcaagtcagg tgacaactcc agttccagtc tgggagatgt ggtgactggc
      601 acccgccgcc ccacaccccc cgccagtgcc aaacagaagc agaagtcgac agagcacgtg
      661 cccccctatg acgtggtgcc ttccatgagg cccatcatcc tggtgggacc atcgctcaag
      721 ggctatgagg taactgacat gatgcagaag gcgttgtttg acttcttgaa gcatcggttt
      781 gatggcagga tttccatcac ccgggtaacg gctgacattt ccctggccaa gcgctcggtc
      841 ctcaacaacc ccagcaaaca cattatcatc gagcgctcca acactcgctc cagcctggct
      901 gaggtacaga gtgaaatcga gcggatcttc gagctggccc ggaccttgca gctggttgcc
      961 ctggacgccg acaccatcaa ccacccagcc cagctctcta aaacctcgct ggcccccatc
     1021 attgtttaca tcaagatcac atcccccaag gtactgcaga ggctcatcaa atcccgaggg
     1081 aagtctcaat ccaaacacct caatgtccaa atagcagcct cggagaagct ggcacagtgt
     1141 ccccccgaaa tgtttgacat aatcctggac gagaaccaat tggaagatgc ctgcgagcac
     1201 ctggcggagt acttggaagc ctactggaag gccacacacc cgcctagcag cacgccaccc
     1261 aatccgctgc tgaaccgcac catggctacc gccgctctgg ctgccagccc tgcccccgtc
     1321 tccaacctcc agggacccta ccttgcttcc ggggaccagc cgctggaccg ggccactggg
     1381 gagcatgcca gtgtgcacga gtaccccggg gagctgggcc agcccccagg cctttacccc
     1441 agcaaccacc cacctggccg ggcaggcacc ctgcgggcgc tatcccgcca agacaccttt
     1501 gatgctgaca cccccggcag ccgaaactct gcctacacgg agccaggaga ctcgtgtgtg
     1561 gacatggaga cagacccctc agagggccca gggcctggag accctgcagg gggaggtaca
     1621 ccaccagctc ggcagggctc ctgggaagag gaggaagatt atgaggagga gatgaccgac
     1681 aacaggaacc ggggccggaa taaggcccgc tactgtgcgg agggtggtgg gccggttctg
     1741 gggcgcaata agaatgagct ggagggctgg ggacaaggcg tctacatccg cactagtgtg
     1801 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     1861 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1921 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     1981 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     2041 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     2101 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     2161 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     2221 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     2281 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     2341 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     2401 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     2461 gagggccgcc actccaccgg cggcatggac gagctgtaca agtagcggcc gcaattccgg
     2521 gggggggggg gggggggaca gctcagggct gcgatttcgc gccaaacttg acggcaatcc
     2581 tagcgtgaag gctggtagga ttttatcccc gctgccatca tggttcgacc attgaactgc
     2641 atcgtcgccg tgtcccaaaa tatggggatt ggcaagaacg gagacctacc ctggcctccg
     2701 ctcaggaacg agttcaagta cttccaaaga atgaccacaa cctcttcagt ggaaggtaaa
     2761 cagaatctgg tgattatggg taggaaaacc tggttctcca ttcctgagaa gaatcgacct
     2821 ttaaaggaca gaattaatat agttctcagt agagaactca aagaaccacc acgaggagct
     2881 cattttcttg ccaaaagttt ggatgatgcc ttaagactta ttgaacaacc ggaattggca
     2941 agtaaagtag acatggtttg gatagtcgga ggcagttctg tttaccagga agccatgaat
     3001 caaccaggcc acctcagact ctttgtgaca aggatcatgc aggaatttga aagtgacacg
     3061 tttttcccag aaattgattt ggggaaatat aaacttctcc cagaataccc aggcgtcctc
     3121 tctgaggtcc aggaggaaaa aggcatcaag tataagtttg aagtctacga gaagaaagac
     3181 taacaggaag atgctttcaa gttctctgct cccctcctaa agctatgcat ttttataaga
     3241 ccatgggact tttgctggct ttagatcata atcagccata ccacatttgt agaggtttta
     3301 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     3361 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3421 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3481 aatgtatctt atcatgtctg gatccccggc caacggtctg gtgacccggc tgcgagagct
     3541 cggtgtacct gagacgcgag taagcccttg agtcaaagac gtagtcgttg caagtccgca
     3601 ccaggtactg atatcccacc aaaaagtgcg gcggcggctg gcggtagagg ggccagcgta
     3661 gggtggccgg ggctccgggg gcgaggtctt ccaacataag gcgatgatat ccgtagatgt
     3721 acctggacat ccaggtgatg ccggcggcgg tggtggaggc gcgcggaaag tcgcggacgc
     3781 ggttccagat gttgcgcagc ggcaaaaagt gctccatggt cgggacgctc tggccggtga
     3841 ggcgtgcgca gtcgttgacg ctctagaccg tgcaaaagga gagcctgtaa gcgggcactc
     3901 ttccgtggtc tggtggataa attcgcaagg gtatcatggc ggacgaccgg ggttcgaacc
     3961 ccggatccgg ccgtccgccg tgatccatgc ggttaccgcc cgcgtgtcga acccaggtgt
     4021 gcgacgtcag acaacggggg agcgctcctt ttggcttcct tccaggcgcg gcggctgctg
     4081 cgctagcttt tttggcgagc tcgaattaat tctgcattaa tgaatcggcc aacgcgcggg
     4141 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4201 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4261 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4321 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4381 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4441 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4501 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4561 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4621 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4681 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4741 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4801 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4861 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4921 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4981 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5041 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5101 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5161 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5221 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5281 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5341 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5401 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5461 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5521 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5581 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5641 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5701 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5761 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5821 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5881 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5941 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6001 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6061 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     6121 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     6181 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     6241 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     6301 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     6361 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     6421 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     6481 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     6541 acgttgtaaa acgacggcca gtgccaagct ttttgcaaaa gcctaggcct ccaaaaaagc
     6601 ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa
     6661 aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga
     6721 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac
     6781 ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca
     6841 tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca
     6901 cattccacag gatccggtcg cgcgaatttc gagcggtgtt ccgcggtcct cctcgtatag
     6961 aaactcggac cactctgaga cgaaggctcg cgtccaggcc agcacgaagg aggctaagtg
     7021 ggaggggtag cggtcgttgt ccactagggg gtccactcgc tccagggtgt gaagacacat
     7081 gtcgccctct tcggcatcaa ggaaggtgat tggtttatag gtgtaggcca cgtgaccggg
     7141 tgttcctgaa ggggggctat aaaagggggt gggggcgcgt tcgtcctcac tctcttccgc
     7201 atcgctgtct gcgagggcca gctgttgggc tcgcggttga ggacaaactc ttcgcggtct
     7261 ttccagtact cttggatcgg aaacccgtcg gcctccgaac ggtactccgc caccgaggga
     7321 cctgagcgag tccgcatcga ccggatcgga aaacctctcg actgttgggg tgagtactcc
     7381 ctctcaaaag cgggcatgac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat
     7441 ttgatattca cctggcccgc ggtgatgcct ttgagggtgg ccgcgtccat ctggtcagaa
     7501 aagacaatct ttttgttgtc aagcttgagg tgtggcaggc ttgagatctg gccatacact
     7561 tgagtgacaa tgacatccac tttgcctttc tctccacagg tgtccactcc caggtccaac
     7621 tgcaggcgag cctgaattcg gcacgagagc gtgcagctgg gcgactgcac ccggtgctgg
     7681 ctgcgcgacg ccgcgctgct ctcggctcgg acggcctctc ccatgcgctg agggcccccg
     7741 gctgtgccgg gagggcggcc ggaccggagg atcctctcc
//
