LOCUS       sequence-228918-        5875 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Plasmid co-producing the LPA2 receptor and a mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5875)
  AUTHORS   Mastop M, Reinhard NR, Zuconelli CR, Terwey F, Gadella TWJ Jr, van 
            Unen J, Adjobo-Hermans MJW, Goedhart J
  TITLE     A FRET-based biosensor for measuring Galpha13 activation in single 
            cells.
  JOURNAL   PLoS One. 2018 Mar 5;13(3):e0193705. doi: 
            10.1371/journal.pone.0193705. eCollection 2018.
  PUBMED    29505611
REFERENCE   2  (bases 1 to 5875)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2018-03-5"; volume: "13(3):e0193705. doi"; pages: " 
            10.1371/journal.pone.0193705. eCollection 2018"
COMMENT     Sequence Label: LPA2R-P2A-mCherry
FEATURES             Location/Qualifiers
     source          1..5875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1767..1823
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1830..2540
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1975..1993)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2243..2262
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2663..2784
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2700..2719)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2754..2773
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2791..3246)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2928..2949)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3140..3159
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3273..3377
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3379..3736
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3379..3570
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3402..3422)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3587..3722
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3771..4565
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3771..4565
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3825..3844)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4435..4454
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4753..4772)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4797..4844
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5173..5761
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5662..5681
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttggt accgagctcg gatccactag tccagtgtgg
      661 tggaattcac catggtcatc atgggccagt gctactacaa cgagaccatc ggcttcttct
      721 ataacaacag tggcaaagag ctcagctccc actggcggcc caaggatgtg gtcgtggtgg
      781 cactggggct gaccgtcagc gtgctggtgc tgctgaccaa tctgctggtc atagcagcca
      841 tcgcctccaa ccgccgcttc caccagccca tctactacct gctcggcaat ctggccgcgg
      901 ctgacctctt cgcgggcgtg gcctacctct tcctcatgtt ccacactggt ccccgcacag
      961 cccgactttc acttgagggc tggttcctgc ggcagggctt gctggacaca agcctcactg
     1021 cgtcggtggc cacactgctg gccatcgccg tggagcggca ccgcagtgtg atggccgtac
     1081 agctgcacag ccgcctgccc cgtggccgcg tggtcatgct cattgtgggc gtgtgggtgg
     1141 ctgccctggg cctggggctg ctgcctgccc actcctggca ctgcctctgt gccctggacc
     1201 gctgctcacg catggcaccc ctgctcagcc gctcctattt ggccgtctgg gctctgtcga
     1261 gcctgcttgt cttcctgctc atggtggctg tgtacacccg cattttcttc tacgtgcggc
     1321 ggcgagtgca gcgcatggca gagcatgtca gctgccaccc ccgctaccga gagaccacgc
     1381 tcagcctggt caagactgtt gtcatcatcc tgggggcgtt cgtggtctgc tggacaccag
     1441 gccaggtggt actgctcctg gatggtttag gctgtgagtc ctgcaatgtc ctggctgtag
     1501 aaaagtactt cctactgttg gccgaggcca actcactggt caatgctgct gtgtactctt
     1561 gccgagatgc tgagatgcgc cgcaccttcc gccgccttct ctgctgcgcg tgcctccgcc
     1621 agtccacccg cgagtctgtc cactatacat cctctgccca gggaggtgcc agcactcgca
     1681 tcatgcttcc cgagaacggc cacccactga tgactccacc caagtcgacg gtaccgcggg
     1741 cccgggatcc accggtggga tcgggtgcta ctaacttcag cctgctgaag caggctggag
     1801 acgtggagga gaaccctgga cctgggtcca tggtgagcaa gggcgaggag gataacatgg
     1861 ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg
     1921 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     1981 tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca
     2041 tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt
     2101 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga
     2161 ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg
     2221 gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct
     2281 cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga
     2341 agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc
     2401 ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg
     2461 aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca
     2521 tggacgagct gtacaagtaa agcggccgcg actctagatc ataatcagcc ataccacatt
     2581 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2641 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     2701 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2761 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     2821 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     2881 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     2941 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3001 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     3061 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3121 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     3181 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     3241 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3301 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3361 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     3421 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3481 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3541 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3601 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3661 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3721 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3781 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3841 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3901 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     3961 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4021 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4081 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4141 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4201 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4261 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     4321 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4381 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4441 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4501 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4561 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4621 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4681 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     4741 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     4801 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     4861 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4921 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     4981 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     5041 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5101 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5161 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5221 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5281 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5341 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5401 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5461 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5521 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5581 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5641 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5701 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5761 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5821 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
