LOCUS       sequence-194168-        6812 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ACP-Tagged beta-2 Adrenergic Receptor in Mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6812)
  AUTHORS   Ana Egana
  TITLE     Egana Lab NEB plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6812)
  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            "Unpublished"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: pACP-ADR-beta-2 Control Plasmid
FEATURES             Location/Qualifiers
     source          1..6812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    105..567
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(161..178)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     388..407
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             603..1406
                     /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(666..685)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1276..1295
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1536..1657
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1573..1592)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1759..1775)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1759..1775)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1772..1794)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1783..1799
                     /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(1807..1813,1814..1831,1832..1837))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1852..1873
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1990..2007)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2161..2749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2241..2260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2920..3711,3712..3780))
                     /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     3543..3562
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3781..3885)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3960..3979)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4151..4530
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4531..4734
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4685..4705
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4779..4798
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4779..4797
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4927..5160
                     /codon_start=1
                     /gene="acpP"
                     /product="acyl carrier protein (ACP) from E. coli"
                     /label=ACP
                     /translation="MSTIEERVKKIIGEQLGVKQEEVTNNASFVEDLGADSLDTVELVM
                     ALEEEFDTEIPDEEAEKITTVQAAIDYINGHQA"
     intron          6516..6745
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     6728..6747
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 cccccccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
       61 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
      121 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
      181 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
      241 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
      301 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
      361 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
      421 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
      481 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
      541 gttttccttt gaaaaacacg atgataagct tgccacaacc cgggataatt cctgcagcca
      601 atatgggatc ggccattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
      661 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
      721 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
      781 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
      841 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
      901 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
      961 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     1021 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     1081 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     1141 gcatgcccga cggcgatgat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     1201 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     1261 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     1321 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     1381 atcgccttct tgacgagttc ttctgagggg atcaattctc tagataactg atcataatca
     1441 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     1501 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     1561 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     1621 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtcgagtgc attctagttg
     1681 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     1741 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     1801 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     1861 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1921 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1981 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2041 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2101 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2161 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2221 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2281 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2341 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2401 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2461 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2521 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2581 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2641 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2701 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2761 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2821 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2881 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2941 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3001 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3061 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3121 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3181 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3241 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3301 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3361 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3421 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3481 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3541 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3601 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3661 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3721 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3781 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3841 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3901 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     3961 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     4021 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     4081 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     4141 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     4201 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     4261 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     4321 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     4381 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     4441 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     4501 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     4561 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     4621 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     4681 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
     4741 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
     4801 acccaagctt ggtaccgagc tcggatcgtc gctagcacca tgcggctctg catcccgcag
     4861 gtgctgttgg ccttgttcct ttccatgctg acagggccgg gagaaggcag cgaattcaag
     4921 cttaccatga gcactatcga agaacgcgtt aagaaaatta tcggcgaaca gctgggcgtt
     4981 aagcaggaag aagttaccaa caatgcttct ttcgttgaag acctgggcgc ggattctctt
     5041 gacaccgttg agctggtaat ggctctggaa gaagagtttg atactgagat tccggacgaa
     5101 gaagctgaga aaatcaccac cgttcaggct gccattgatt acatcaacgg ccaccaggcg
     5161 cctgcaggta ccatggggca acccgggaac ggcagcgcct tcttgctggc acccaatgga
     5221 agccatgcgc cggaccacga cgtcacgcag caaagggacg aggtgtgggt ggtgggcatg
     5281 ggcatcgtca tgtctctcat cgtcctggcc atcgtgtttg gcaatgtgct ggtcatcaca
     5341 gccattgcca agttcgagcg tctgcagacg gtcaccaact acttcatcac ttcactggcc
     5401 tgtgctgatc tggtcatggg cctagcagtg gtgccctttg gggccgccca tattcttatg
     5461 aaaatgtgga cttttggcaa cttctggtgc gagttttgga cttccattga tgtgctgtgc
     5521 gtcacggcca gcattgagac cctgtgcgtg atcgcagtgg atcgctactt tgccattact
     5581 tcacctttca agtaccagag cctgctgacc aagaataagg cccgggtgat cattctgatg
     5641 gtgtggattg tgtcaggcct tacctccttc ttgcccattc agatgcactg gtacagggcc
     5701 acccaccagg aagccatcaa ctgctatgcc aatgagacct gctgtgactt cttcacgaac
     5761 caagcctatg ccattgcctc ttccatcgtg tccttctacg ttcccctggt gatcatggtc
     5821 ttcgtctact ccagggtctt tcaggaggcc aaaaggcagc tccagaagat tgacaaatct
     5881 gagggccgct tccatgtcca gaaccttagc caggtggagc aggatgggcg gacggggcat
     5941 ggactccgca gatcttccaa gttctgcttg aaggagcaca aagccctcaa gacgttaggc
     6001 atcatcatgg gcactttcac cctctgctgg ctgcccttct tcatcgttaa cattgtgcat
     6061 gtgatccagg ataacctcat ccgtaaggaa gtttacatcc tcctaaattg gataggctat
     6121 gtcaattctg gtttcaatcc ccttatctac tgccggagcc cagatttcag gattgccttc
     6181 caggagcttc tgtgcctgcg caggtcttct ttgaaggcct atggcaatgg ctactccagc
     6241 aacggcaaca caggggagca gagtggatat cacgtggaac aggagaaaga aaataaactg
     6301 ctgtgtgaag acctcccagg cacggaagac tttgtgggcc atcaaggtac tgtgcctagc
     6361 gataacattg attcacaagg gaggaattgt agtacaaatg actcactgct gtaaggcgcg
     6421 ccaggatccc tcgaggttaa ttaatgagcg gccgcataga taactgatcc agtgtgctgg
     6481 aattaattcg ctgtctgcga gggccagctg ttggggtgag tactccctct caaaagcggg
     6541 catgacttct gcgctaagat tgtcagtttc caaaaacgag gaggatttga tattcacctg
     6601 gcccgcggtg atgcctttga gggtggccgc gtccatctgg tcagaaaaga caatcttttt
     6661 gttgtcaagc ttgaggtgtg gcaggcttga gatctggcca tacacttgag tgacaatgac
     6721 atccactttg cctttctctc cacaggtgtc cactcccagg tccaactgca ggtcgagcat
     6781 gcatctaggg cggccaattc cgcccctctc cc
//
