LOCUS       sequence-182021-        6792 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of CD86-mEos2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6792)
  AUTHORS   Fricke F, Beaudouin J, Eils R, Heilemann M
  TITLE     One, two or three? Probing the stoichiometry of membrane proteins by
            single-molecule localization microscopy.
  JOURNAL   Sci Rep. 2015 Sep 11;5:14072. doi: 10.1038/srep14072.
  PUBMED    26358640
REFERENCE   2  (bases 1 to 6792)
  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: "Sci Rep."; 
            date: "2015-09-11"; volume: "5:14072. doi"; pages: " 
            10.1038/srep14072"
COMMENT     Sequence Label: pirespuro2-CD86-mEos2
FEATURES             Location/Qualifiers
     source          1..6792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        24..403
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        404..607
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     558..578
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      654..663
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          2442..2671
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2654..2673
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2729..3300
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2894..2911)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3121..3140
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3333..3932
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3333..3352)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3829..3849
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(4068..4085)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4074..4298
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4345..4364
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4424..4440)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4424..4440)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4437..4459)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4448..4464
                     /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(4472..4478,4479..4496,4497..4502))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4517..4538
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4655..4672)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4826..5414)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4906..4925)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5585..6376,6377..6445))
                     /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     6208..6227
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6446..6550)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6625..6644)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
       61 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      121 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      181 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      241 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      301 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      361 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      421 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      481 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      541 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      601 cagagctggt ttagtgaacc gtcagatccg ctagctaatc tcgagatgct agcgccacca
      661 tgggactgag taacattctc tttgtgatgg ccttcctgct ctctggtgct gctcctctga
      721 agattcaagc ttatttcaat gagactgcag acctgccatg ccaatttgca aactctcaaa
      781 accaaagcct gagtgagcta gtagtatttt ggcaggacca ggaaaacttg gttctgaatg
      841 aggtatactt aggcaaagag aaatttgaca gtgttcattc caagtatatg ggccgcacaa
      901 gttttgattc ggacagttgg accctgagac ttcacaatct tcagatcaag gacaagggct
      961 tgtatcaatg tatcatccat cacaaaaagc ccacaggaat gattcgcatc caccagatga
     1021 attctgaact gtcagtgctt gctaacttca gtcaacctga aatagtacca atttctaata
     1081 taacagaaaa tgtgtacata aatttgacct gctcatctat acacggttac ccagaaccta
     1141 agaagatgag tgttttgcta agaaccaaga attcaactat cgagtatgat ggtattatgc
     1201 agaaatctca agataatgtc acagaactgt acgacgtttc catcagcttg tctgtttcat
     1261 tccctgatgt tacgagcaat atgaccatct tctgtattct ggaaactgac aagacgcggc
     1321 ttttatcttc acctttctct atagagcttg aggaccctca gcctccccca gaccacattc
     1381 cttggattac agctgtactt ccaacagtta ttatatgtgt gatggttttc tgtctaattc
     1441 tatggaaatg gaagaagaag aagcggcctc gcaactctta taaatgtgga accaacacaa
     1501 tggagaggga agagagtgaa cagaccaaga aaagagaaaa aatccatata cctgaaagat
     1561 ctgatgaaac ccagcgtgtt tttaaaagtt cgaagacatc ttcatgcgac aaaagtgata
     1621 catgttttgg aggcggaccg gtgccccagt gggagggatt cgctgcgctg ctggcaaccc
     1681 cggtcgctac tatgtctgct atcaagccag acatgaagat caagttgaga atggaaggta
     1741 acgttaacgg tcaccacttc gttatcgacg gtgacggtac tggtaagcca ttcgaaggta
     1801 agcaatctat ggacttggaa gttaaggaag gtggtccatt gccattcgct ttcgacatct
     1861 tgactactgc tttccactac ggtaacagag ttttcgctaa gtacccagac aacatccaag
     1921 actacttcaa gcaatctttc ccaaagggtt actcttggga aagatctttg actttcgaag
     1981 acggtggtat ctgtatcgct agaaacgaca tcactatgga aggtgacact ttctacaaca
     2041 aggttagatt ctacggtact aacttcccag ctaacggtcc agttatgcaa aagaagactt
     2101 tgaagtggga accatctact gaaaagatgt acgttagaga cggtgttttg actggtgaca
     2161 tccacatggc tttgttgttg gaaggtaacg ctcactacag atgtgacttc agaactactt
     2221 acaaggctaa ggaaaagggt gttaagttgc caggttacca cttcgttgac cactgtatcg
     2281 aaatcttgtc tcacgacaag gactacaaca aggttaagtt gtacgaacac gctgttgctc
     2341 actctggttt gccagacaac gctagaagat aaagcggccg catagataac tgatccagtg
     2401 tgctggaatt aattcgctgt ctgcgagggc cagctgttgg ggtgagtact ccctctcaaa
     2461 agcgggcatg acttctgcgc taagattgtc agtttccaaa aacgaggagg atttgatatt
     2521 cacctggccc gcggtgatgc ctttgagggt ggccgcgtcc atctggtcag aaaagacaat
     2581 ctttttgttg tcaagcttga ggtgtggcag gcttgagatc tggccataca cttgagtgac
     2641 aatgacatcc actttgcctt tctctccaca ggtgtccact cccaggtcca actgcaggtc
     2701 gagcatgcat ctagggcggc caattccgcc cctctccccc ccccccctaa cgttactggc
     2761 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     2821 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     2881 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     2941 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     3001 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     3061 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     3121 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     3181 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     3241 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     3301 gcttgccaca acccacaagg agacgacctt ccatgaccga gtacaagccc acggtgcgcc
     3361 tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact
     3421 accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc
     3481 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     3541 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     3601 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     3661 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     3721 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     3781 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     3841 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     3901 cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga
     3961 ccgaaaggag cgcacgaccc catggctccg accgaagcca cccggggcgg ccccgccgac
     4021 cccgcacccg cccccgaggc ccaccgactc tagagctcgc tgatcagcct cgactgtgcc
     4081 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     4141 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     4201 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4261 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     4321 ctggggctcg agtgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4381 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     4441 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4501 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4561 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4621 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4681 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4741 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4801 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4861 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4921 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4981 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5041 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5101 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5161 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5221 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5281 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5341 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5401 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5461 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5521 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5581 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5641 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5701 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5761 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5821 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5881 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5941 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6001 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6061 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6121 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6181 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6241 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6301 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6361 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6421 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6481 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6541 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     6601 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     6661 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     6721 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     6781 gctgcttcgc ga
//
