LOCUS       sequence-223582-        5823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid co-producing the AT1R receptor and a mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5823)
  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 5823)
  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: "PLoS One.";
            date: "2018-03-5"; volume: "13(3):e0193705. doi"; pages: " 
            10.1371/journal.pone.0193705. eCollection 2018"
COMMENT     Sequence Label: At1R-P2A-mCherry
FEATURES             Location/Qualifiers
     source          1..5823
                     /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             1715..1771
                     /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             1778..2488
                     /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(1923..1941)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2191..2210
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2611..2732
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2648..2667)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2702..2721
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2739..3194)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2876..2897)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3088..3107
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3221..3325
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3327..3684
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3327..3518
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3350..3370)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3535..3670
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3719..4513
                     /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             3719..4513
                     /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(3773..3792)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4383..4402
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4701..4720)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4745..4792
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5121..5709
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5610..5629
                     /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 caagcttcaa aatggccctt aactcttctg ctgaagatgg
      661 tatcaaaaga atccaagatg actgccccaa ggctggcagg cacagttaca tatttgtcat
      721 gatccctacc ctctacagca tcatctttgt ggtgggaata tttggaaaca gcttggtggt
      781 gattgtcatt tacttttaca tgaagctgaa gactgtggcc agcgtctttc ttctcaatct
      841 cgccttggct gacttatgct ttttgctgac tttgcccctg tgggcagtct ataccgctat
      901 ggagtaccgc tggcccttcg gcaatcacct atgtaagatc gcttcggcca gcgtgagctt
      961 caacctctac gccagtgtgt tccttctcac gtgtctcagc atcgaccgct acctggccat
     1021 cgtccaccca atgaagtctc gccttcgccg cacgatgctg gtggccaaag tcacctgcat
     1081 catcatctgg ctgatggctg gcttggccag tttgccagct gtcatccacc gaaatgtata
     1141 cttcatcgag aacaccaata tcacagtgtg cgcgtttcat tatgagtctc ggaattcgac
     1201 gctccccata gggctgggcc ttaccaagaa tattctgggc ttcttgttcc ctttccttat
     1261 cattctcacc agctataccc ttatttggaa agctctaaag aaggcttatg aaattcaaaa
     1321 gaacaaacca agaaacgatg acatctttag gataattatg gcgattgtgc ttttcttctt
     1381 cttttcctgg gtcccccacc aaatattcac tttcctggat gtgctgattc agctgggcgt
     1441 catccatgac tgtaaaattt ctgacatcgt ggacactgcc atgcccatca ccatctgcat
     1501 agcgtatttt aacaactgcc tgaaccctct gttctacggc tttctgggga agaaatttaa
     1561 aaagtatttc ctccagctcc tgaaatatat tcccccaaag gccaagtccc actcaagcct
     1621 gtctacgaaa atgagcacgc tttcttaccg gccttcggat aacatgagct catcggccaa
     1681 aaagcctgcg tcttgttttg aggtggaacc ggtggctact aacttcagcc tgctgaagca
     1741 ggctggagac gtggaggaga accctggacc tgggtccatg gtgagcaagg gcgaggagga
     1801 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
     1861 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     1921 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
     1981 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
     2041 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     2101 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     2161 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     2221 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
     2281 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
     2341 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
     2401 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     2461 cggcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat
     2521 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     2581 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     2641 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     2701 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt
     2761 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     2821 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     2881 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     2941 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     3001 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     3061 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     3121 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     3181 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3241 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3301 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     3361 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3421 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3481 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3541 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3601 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     3661 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     3721 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3781 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3841 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3901 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3961 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4021 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4081 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4141 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4201 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     4261 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4321 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4381 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4441 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4501 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4561 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4621 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4681 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     4741 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     4801 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     4861 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     4921 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     4981 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5041 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5101 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5161 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5221 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5281 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5341 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5401 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5461 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5521 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5581 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5641 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5701 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5761 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     5821 cat
//
