LOCUS       Exported                5945 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Visualization of the Beta-2-adrenergic receptor.
ACCESSION   .
VERSION     .
KEYWORDS    HsB2AR-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5945)
  AUTHORS   Pietraszewska-Bogiel A, Joosen L, Chertkova AO, Goedhart J
  TITLE     Not So Dry After All: DRY Mutants of the AT1A Receptor and H1 
            Receptor Can Induce G-Protein-Dependent Signaling.
  JOURNAL   ACS Omega. 2020 Feb 3;5(6):2648-2659. doi: 10.1021/acsomega.9b03146.
            eCollection 2020 Feb 18.
  PUBMED    32095688
REFERENCE   2  (bases 1 to 5945)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5945
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1834..1843
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1840..2550
                     /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(1985..2003)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2253..2272
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2673..2794
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2710..2729)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2764..2783
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2801..3256)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2938..2959)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3150..3169
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3283..3387
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3389..3746
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3412..3432)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3597..3732
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3659..3678
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3781..4575
                     /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(3835..3854)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4445..4464
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4763..4782)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4807..4854
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5183..5771
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5672..5691
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcgattat ccaccatggg gcaacccggg
      601 aacggcagcg ccttcttgct ggcacccaat agaagccatg cgccggacca cgacgtcacg
      661 cagcaaaggg acgaggtgtg ggtggtgggc atgggcatcg tcatgtctct catcgtcctg
      721 gccatcgtgt ttggcaatgt gctggtcatc acagccattg ccaagttcga gcgtctgcag
      781 acggtcacca actacttcat cacttcactg gcctgtgctg atctggtcat gggcctggca
      841 gtggtgccct ttggggccgc ccatattctt atgaaaatgt ggacttttgg caacttctgg
      901 tgcgagtttt ggacttccat tgatgtgctg tgcgtcacgg ccagcattga gaccctgtgc
      961 gtgatcgcag tggatcgcta ctttgccatt acttcacctt tcaagtacca gagcctgctg
     1021 accaagaata aggcccgggt gatcattctg atggtgtgga ttgtgtcagg ccttacctcc
     1081 ttcttgccca ttcagatgca ctggtaccgg gccacccacc aggaagccat caactgctat
     1141 gccaatgaga cctgctgtga cttcttcacg aaccaagcct atgccattgc ctcttccatc
     1201 gtgtccttct acgttcccct ggtgatcatg gtcttcgtct actccagggt ctttcaggag
     1261 gccaaaaggc agctccagaa gattgacaaa tctgagggcc gcttccatgt ccagaacctt
     1321 agccaggtgg agcaggatgg gcggacgggg catggactcc gcagatcttc caagttctgc
     1381 ttgaaggagc acaaagccct caagacgtta ggcatcatca tgggcacttt caccctctgc
     1441 tggctgccct tcttcatcgt taacattgtg catgtgatcc aggataacct catccgtaag
     1501 gaagtttaca tcctcctaaa ttggataggc tatgtcaatt ctggtttcaa tccccttatc
     1561 tactgccgga gcccagattt caggattgcc ttccaggagc ttctgtgcct gcgcaggtct
     1621 tctttgaagg cctatgggaa tggctactcc agcaacggca acacagggga gcagagtgga
     1681 tatcacgtgg aacaggagaa agaaaataaa ctgctgtgtg aagacctccc aggcacggaa
     1741 gactttgtgg gccatcaagg tactgtgcct agcgataaca ttgattcaca agggaggaat
     1801 tgtagtacaa atgactcact gctgggaccg gtcgccacca tggtgagcaa gggcgaggag
     1861 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     1921 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     1981 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     2041 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     2101 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     2161 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     2221 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     2281 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     2341 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     2401 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     2461 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     2521 accggcggca tggacgagct gtacaagtaa agcggccgcg actctagatc ataatcagcc
     2581 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     2641 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     2701 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2761 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     2821 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     2881 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     2941 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3001 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     3061 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3121 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3181 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3241 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3301 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3361 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     3421 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3481 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3541 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3601 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3661 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3721 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     3781 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3841 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3901 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3961 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4021 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4081 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4141 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4201 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4261 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     4321 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4381 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4441 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4501 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4561 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4621 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4681 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4741 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     4801 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     4861 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     4921 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     4981 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     5041 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5101 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5161 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5221 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5281 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5341 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5401 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5461 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5521 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5581 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5641 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5701 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5761 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5821 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     5881 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     5941 ttccg
//
