LOCUS       sequence-194092-        7164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SNAP-Tagged beta-2 Adrenergic Receptor in Mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7164)
  AUTHORS   Ana Egana
  TITLE     Egana Lab NEB plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7164)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pSNAPf-ADR-beta-2 Control Plasmid
FEATURES             Location/Qualifiers
     source          1..7164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(167..184)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     394..413
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             609..1412
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(672..691)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1282..1301
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1542..1663
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1579..1598)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1765..1781)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1765..1781)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1778..1800)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1789..1805
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1813..1819,1820..1837,1838..1843))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1858..1879
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1996..2013)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2167..2755)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2247..2266)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2926..3717,3718..3786))
                     /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     3549..3568
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3787..3891)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3966..3985)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4157..4536
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4537..4740
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4691..4711
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4785..4804
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4785..4803
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4969..5514
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     intron          6862..7091
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     7074..7093
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(7164,1..574,575..577,578..586)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
ORIGIN
        1 ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
       61 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
      121 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
      181 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
      241 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
      301 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
      361 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
      421 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc
      481 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
      541 gacgtggttt tcctttgaaa aacacgatga taagcttgcc acaacccggg ataattcctg
      601 cagccaatat gggatcggcc attgaacaag atggattgca cgcaggttct ccggccgctt
      661 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
      721 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
      781 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
      841 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
      901 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
      961 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     1021 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     1081 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     1141 aggcgcgcat gcccgacggc gatgatctcg tcgtgaccca tggcgatgcc tgcttgccga
     1201 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     1261 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     1321 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     1381 ccttctatcg ccttcttgac gagttcttct gaggggatca attctctaga taactgatca
     1441 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     1501 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     1561 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     1621 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgtc gagtgcattc
     1681 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     1741 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     1801 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     1861 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     1921 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1981 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2041 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2101 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2161 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2221 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2281 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2341 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2401 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2461 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2521 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2581 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2641 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2701 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2761 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2821 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2881 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2941 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3001 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3061 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3121 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3181 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3241 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3301 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3361 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3421 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3481 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3541 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3601 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3661 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3721 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3781 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3841 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3901 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     3961 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     4021 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     4081 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
     4141 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
     4201 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     4261 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     4321 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     4381 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     4441 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     4501 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     4561 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     4621 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     4681 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     4741 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat
     4801 agggagaccc aagcttggta ccgagctcgg atcgatatca ccatgcggct ctgcatcccg
     4861 caggtgctgt tggccttgtt cctttccatg ctgacagggc cgggagaagg cagcgctagc
     4921 gatatcggcg cgccagcatt taaatctgta cagaccggtg aattcaccat ggacaaagac
     4981 tgcgaaatga agcgcaccac cctggatagc cctctgggca agctggaact gtctgggtgc
     5041 gaacagggcc tgcaccgtat catcttcctg ggcaaaggaa catctgccgc cgacgccgtg
     5101 gaagtgcctg ccccagccgc cgtgctgggc ggaccagagc cactgatgca ggccaccgcc
     5161 tggctcaacg cctactttca ccagcctgag gccatcgagg agttccctgt gccagccctg
     5221 caccacccag tgttccagca ggagagcttt acccgccagg tgctgtggaa actgctgaaa
     5281 gtggtgaagt tcggagaggt catcagctac agccacctgg ccgccctggc cggcaatccc
     5341 gccgccaccg ccgccgtgaa aaccgccctg agcggaaatc ccgtgcccat tctgatcccc
     5401 tgccaccggg tggtgcaggg cgacctggac gtggggggct acgagggcgg gctcgccgtg
     5461 aaagagtggc tgctggccca cgagggccac agactgggca agcctgggct gggtcctgca
     5521 ggtaccatgg ggcaacccgg gaacggcagc gccttcttgc tggcacccaa tggaagccat
     5581 gcgccggacc acgacgtcac gcagcaaagg gacgaggtgt gggtggtggg catgggcatc
     5641 gtcatgtctc tcatcgtcct ggccatcgtg tttggcaatg tgctggtcat cacagccatt
     5701 gccaagttcg agcgtctgca gacggtcacc aactacttca tcacttcact ggcctgtgct
     5761 gatctggtca tgggcctagc agtggtgccc tttggggccg cccatattct tatgaaaatg
     5821 tggacttttg gcaacttctg gtgcgagttt tggacttcca ttgatgtgct gtgcgtcacg
     5881 gccagcattg agaccctgtg cgtgatcgca gtggatcgct actttgccat tacttcacct
     5941 ttcaagtacc agagcctgct gaccaagaat aaggcccggg tgatcattct gatggtgtgg
     6001 attgtgtcag gccttacctc cttcttgccc attcagatgc actggtacag ggccacccac
     6061 caggaagcca tcaactgcta tgccaatgag acctgctgtg acttcttcac gaaccaagcc
     6121 tatgccattg cctcttccat cgtgtccttc tacgttcccc tggtgatcat ggtcttcgtc
     6181 tactccaggg tctttcagga ggccaaaagg cagctccaga agattgacaa atctgagggc
     6241 cgcttccatg tccagaacct tagccaggtg gagcaggatg ggcggacggg gcatggactc
     6301 cgcagatctt ccaagttctg cttgaaggag cacaaagccc tcaagacgtt aggcatcatc
     6361 atgggcactt tcaccctctg ctggctgccc ttcttcatcg ttaacattgt gcatgtgatc
     6421 caggataacc tcatccgtaa ggaagtttac atcctcctaa attggatagg ctatgtcaat
     6481 tctggtttca atccccttat ctactgccgg agcccagatt tcaggattgc cttccaggag
     6541 cttctgtgcc tgcgcaggtc ttctttgaag gcctatggca atggctactc cagcaacggc
     6601 aacacagggg agcagagtgg atatcacgtg gaacaggaga aagaaaataa actgctgtgt
     6661 gaagacctcc caggcacgga agactttgtg ggccatcaag gtactgtgcc tagcgataac
     6721 attgattcac aagggaggaa ttgtagtaca aatgactcac tgctgtaagg cgcgccagga
     6781 tccctcgagt gaggcggccg catagataac tgatccagtg tgctggaatt aattcgctgt
     6841 ctgcgagggc cagctgttgg ggtgagtact ccctctcaaa agcgggcatg acttctgcgc
     6901 taagattgtc agtttccaaa aacgaggagg atttgatatt cacctggccc gcggtgatgc
     6961 ctttgagggt ggccgcgtcc atctggtcag aaaagacaat ctttttgttg tcaagcttga
     7021 ggtgtggcag gcttgagatc tggccataca cttgagtgac aatgacatcc actttgcctt
     7081 tctctccaca ggtgtccact cccaggtcca actgcaggtc gagcatgcat ctagggcggc
     7141 caattccgcc cctctccccc cccc
//
