LOCUS       sequence-176909-        7783 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression of Beta1b GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7783)
  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 7783)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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 GFP
FEATURES             Location/Qualifiers
     source          1..7783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1795..2505
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1822..1850)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2445..2465
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2624..3187
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    3374..3508
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3411..3430)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3465..3484
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4181..4198)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4352..4940)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4432..4451)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5111..5902,5903..5971))
                     /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     5734..5753
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5972..6076)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6144..6162
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6200..6222)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6322..6341
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6535..6557
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6549..6566
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6550..6566
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6576..6905)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6590..6725
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6890..6910
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     intron          7374..7603
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     7586..7605
                     /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 cactagtaaa
     1801 ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat
     1861 gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc
     1921 cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact tgtcactact
     1981 ttcggttatg gtgttcaatg ctttgcgaga tacccagatc atatgaaaca gcatgacttt
     2041 ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatattttt caaagatgac
     2101 gggaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc
     2161 gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa attggaatac
     2221 aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg aatcaaagtt
     2281 aacttcaaaa ttagacacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa
     2341 caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca
     2401 caatctgccc tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt
     2461 gtaacagctg ctgggattac acatggcatg gatgaactat acaaataagc ggccgcaatt
     2521 ccgggggggg gggggggggg gacagctcag ggctgcgatt tcgcgccaaa cttgacggca
     2581 atcctagcgt gaaggctggt aggattttat ccccgctgcc atcatggttc gaccattgaa
     2641 ctgcatcgtc gccgtgtccc aaaatatggg gattggcaag aacggagacc taccctggcc
     2701 tccgctcagg aacgagttca agtacttcca aagaatgacc acaacctctt cagtggaagg
     2761 taaacagaat ctggtgatta tgggtaggaa aacctggttc tccattcctg agaagaatcg
     2821 acctttaaag gacagaatta atatagttct cagtagagaa ctcaaagaac caccacgagg
     2881 agctcatttt cttgccaaaa gtttggatga tgccttaaga cttattgaac aaccggaatt
     2941 ggcaagtaaa gtagacatgg tttggatagt cggaggcagt tctgtttacc aggaagccat
     3001 gaatcaacca ggccacctca gactctttgt gacaaggatc atgcaggaat ttgaaagtga
     3061 cacgtttttc ccagaaattg atttggggaa atataaactt ctcccagaat acccaggcgt
     3121 cctctctgag gtccaggagg aaaaaggcat caagtataag tttgaagtct acgagaagaa
     3181 agactaacag gaagatgctt tcaagttctc tgctcccctc ctaaagctat gcatttttat
     3241 aagaccatgg gacttttgct ggctttagat cataatcagc cataccacat ttgtagaggt
     3301 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     3361 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3421 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3481 catcaatgta tcttatcatg tctggatccc cggccaacgg tctggtgacc cggctgcgag
     3541 agctcggtgt acctgagacg cgagtaagcc cttgagtcaa agacgtagtc gttgcaagtc
     3601 cgcaccaggt actgatatcc caccaaaaag tgcggcggcg gctggcggta gaggggccag
     3661 cgtagggtgg ccggggctcc gggggcgagg tcttccaaca taaggcgatg atatccgtag
     3721 atgtacctgg acatccaggt gatgccggcg gcggtggtgg aggcgcgcgg aaagtcgcgg
     3781 acgcggttcc agatgttgcg cagcggcaaa aagtgctcca tggtcgggac gctctggccg
     3841 gtgaggcgtg cgcagtcgtt gacgctctag accgtgcaaa aggagagcct gtaagcgggc
     3901 actcttccgt ggtctggtgg ataaattcgc aagggtatca tggcggacga ccggggttcg
     3961 aaccccggat ccggccgtcc gccgtgatcc atgcggttac cgcccgcgtg tcgaacccag
     4021 gtgtgcgacg tcagacaacg ggggagcgct ccttttggct tccttccagg cgcggcggct
     4081 gctgcgctag cttttttggc gagctcgaat taattctgca ttaatgaatc ggccaacgcg
     4141 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4201 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4261 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4321 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4381 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4441 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4501 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4561 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4621 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4681 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4741 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4801 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4861 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4921 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4981 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5041 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5101 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5161 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5221 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5281 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5341 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5401 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5461 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5521 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5581 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5641 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5701 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5761 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5821 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5881 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5941 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6001 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6061 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     6121 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     6181 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     6241 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     6301 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     6361 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     6421 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     6481 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     6541 cacgacgttg taaaacgacg gccagtgcca agctttttgc aaaagcctag gcctccaaaa
     6601 aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa
     6661 ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc
     6721 gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc
     6781 atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga
     6841 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga
     6901 cacacattcc acaggatccg gtcgcgcgaa tttcgagcgg tgttccgcgg tcctcctcgt
     6961 atagaaactc ggaccactct gagacgaagg ctcgcgtcca ggccagcacg aaggaggcta
     7021 agtgggaggg gtagcggtcg ttgtccacta gggggtccac tcgctccagg gtgtgaagac
     7081 acatgtcgcc ctcttcggca tcaaggaagg tgattggttt ataggtgtag gccacgtgac
     7141 cgggtgttcc tgaagggggg ctataaaagg gggtgggggc gcgttcgtcc tcactctctt
     7201 ccgcatcgct gtctgcgagg gccagctgtt gggctcgcgg ttgaggacaa actcttcgcg
     7261 gtctttccag tactcttgga tcggaaaccc gtcggcctcc gaacggtact ccgccaccga
     7321 gggacctgag cgagtccgca tcgaccggat cggaaaacct ctcgactgtt ggggtgagta
     7381 ctccctctca aaagcgggca tgacttctgc gctaagattg tcagtttcca aaaacgagga
     7441 ggatttgata ttcacctggc ccgcggtgat gcctttgagg gtggccgcgt ccatctggtc
     7501 agaaaagaca atctttttgt tgtcaagctt gaggtgtggc aggcttgaga tctggccata
     7561 cacttgagtg acaatgacat ccactttgcc tttctctcca caggtgtcca ctcccaggtc
     7621 caactgcagg cgagcctgaa ttcggcacga gagcgtgcag ctgggcgact gcacccggtg
     7681 ctggctgcgc gacgccgcgc tgctctcggc tcggacggcc tctcccatgc gctgagggcc
     7741 cccggctgtg ccgggagggc ggccggaccg gaggatcctc tcc
//
