LOCUS       Exported                5925 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Beta-2-adrenergic receptor tagged with mRFP on carboxyl terminus.
ACCESSION   .
VERSION     .
KEYWORDS    Beta-2-adrenergic receptor-mRFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5925)
  AUTHORS   Hynes TR, Mervine SM, Yost EA, Sabo JL, Berlot CH
  TITLE     Live cell imaging of Gs and the beta2-adrenergic receptor 
            demonstrates that both alphas and beta1gamma7 internalize upon 
            stimulation and exhibit similar trafficking patterns that differ 
            from that of the beta2-adrenergic receptor.
  JOURNAL   J Biol Chem. 2004 Oct 15;279(42):44101-12. Epub 2004 Aug 5.
  PUBMED    15297467
REFERENCE   2  (bases 1 to 5925)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5925
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             635..658
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1907..1916
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1913..2590
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(2043..2061)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2311..2330
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2505..2528
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2713..2834
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2750..2769)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2804..2823
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2841..3296)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2978..2999)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3190..3209
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3323..3427
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3429..3786
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3452..3472)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3637..3772
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3699..3718
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3821..4615
                     /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(3875..3894)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4485..4504
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4803..4822)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4847..4894
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5223..5811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5712..5731
                     /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 caagcttcga aatggactac aaggacgacg atgacaaggg
      661 gcaacccggg aacggcagcg ccttcttgct ggcacccaat agaagccatg cgccggacca
      721 cgacgtcacg cagcaaaggg acgaggtgtg ggtggtgggc atgggcatcg tcatgtctct
      781 catcgtcctg gccatcgtgt ttggcaatgt gctggtcatc acagccattg ccaagttcga
      841 gcgtctgcag acggtcacca actacttcat cacttcactg gcctgtgctg atctggtcat
      901 gggcctggca gtggtgccct ttggggccgc ccatattctt atgaaaatgt ggacttttgg
      961 caacttctgg tgcgagtttt ggacttccat tgatgtgctg tgcgtcacgg ccagcattga
     1021 gaccctgtgc gtgatcgcag tggatcgcta ctttgccatt acttcacctt tcaagtacca
     1081 gagcctgctg accaagaata aggcccgggt gatcattctg atggtgtgga ttgtgtcagg
     1141 ccttacctcc ttcttgccca ttcagatgca ctggtaccgg gccacccacc aggaagccat
     1201 caactgctat gccaatgaga cctgctgtga cttcttcacg aaccaagcct atgccattgc
     1261 ctcttccatc gtgtccttct acgttcccct ggtgatcatg gtcttcgtct actccagggt
     1321 ctttcaggag gccaaaaggc agctccagaa gattgacaaa tctgagggcc gcttccatgt
     1381 ccagaacctt agccaggtgg agcaggatgg gcggacgggg catggactcc gcagatcttc
     1441 caagttctgc ttgaaggagc acaaagccct caagacgtta ggcatcatca tgggcacttt
     1501 caccctctgc tggctgccct tcttcatcgt taacattgtg catgtgatcc aggataacct
     1561 catccgtaag gaagtttaca tcctcctaaa ttggataggc tatgtcaatt ctggtttcaa
     1621 tccccttatc tactgccgga gcccagattt caggattgcc ttccaggagc ttctgtgcct
     1681 gcgcaggtct tctttgaagg cctatgggaa tggctactcc agcaacggca acacagggga
     1741 gcagagtgga tatcacgtgg aacaggagaa agaaaataaa ctgctgtgtg aagacctccc
     1801 aggcacggaa gactttgtgg gccatcaagg tactgtgcct agcgataaca ttgattcaca
     1861 agggaggaat tgtagtacaa atgactcact gctggatcca ccggtcgcca ccatggcctc
     1921 ctccgaggac gtcatcaagg agttcatgcg cttcaaggtg cgcatggagg gctccgtgaa
     1981 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     2041 cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc
     2101 tcagttccag tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
     2161 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     2221 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     2281 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     2341 ggaggcctcc accgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagat
     2401 gaggctgaag ctgaaggacg gcggccacta cgacgccgag gtcaagacca cctacatggc
     2461 caagaagccc gtgcagctgc ccggcgccta caagaccgac atcaagctgg acatcacctc
     2521 ccacaacgag gactacacca tcgtggaaca gtacgagcgc gccgagggcc gccactccac
     2581 cggcgcctaa agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt
     2641 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     2701 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2761 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2821 atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     2881 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2941 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     3001 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     3061 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     3121 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     3181 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3241 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     3301 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3361 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3421 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     3481 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3541 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3601 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3661 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3721 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3781 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3841 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3901 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3961 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     4021 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4081 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4141 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4201 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4261 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4321 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     4381 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4441 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4501 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4561 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4621 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4681 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4741 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     4801 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4861 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     4921 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     4981 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     5041 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     5101 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5161 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5221 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5281 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5341 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     5401 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5461 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5521 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5581 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5641 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5701 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5761 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5821 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5881 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
