LOCUS       sequence-409218-        6098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of 1M-Qt-FLAG EMcapsulins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6098)
  AUTHORS   Sigmund F, Berezin O, Beliakova S, Magerl B, Drawitsch M, Piovesan 
            A, Goncalves F, Bodea SV, Winkler S, Bousraou Z, Grosshauser M, 
            Samara E, Pujol-Marti J, Schadler S, So C, Irsen S, Walch A, Kofler 
            F, Piraud M, Kornfeld J, Briggman K, Westmeyer GG
  TITLE     Genetically encoded barcodes for correlative volume electron 
            microscopy.
  JOURNAL   Nat Biotechnol. 2023 Apr 17. doi: 10.1038/s41587-023-01713-y.
  PUBMED    37069313
REFERENCE   2  (bases 1 to 6098)
  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: "Nat 
            Biotechnol. 2023 Apr 17. doi: 10.1038/s41587-023-01713-y."
COMMENT     Sequence Label: 1M-Qt-FLAG
FEATURES             Location/Qualifiers
     source          1..6098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      958..967
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2035..2058
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2106..2123)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2112..2336
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2382..2810
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2469..2488)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2679..2700
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2819..2839)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2824..3153
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3004..3139
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3066..3085
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3201..3248
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3267..3641
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3771..3892
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3808..3827)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3862..3881
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3941..3957)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3941..3957)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3954..3976)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3965..3981
                     /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(3989..3995,3996..4013,4014..4019))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4034..4055
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4172..4189)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4343..4931)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4423..4442)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5102..5893,5894..5962))
                     /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     5725..5744
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5963..6067)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggacc ctgagacatg cccttgtcca acaggcggca gctgcacctg tagcgacaag
     1021 tgcaagtgca aaggctgcaa gtgtaccaac tgcaagaagt cctgctgcag ctgctgtcct
     1081 gccggctgtg aaaagtgcgc caaggactgc gtgtgcaagg gcgaagaagg cgccaaagcc
     1141 gaggccgaga agtgctcttg ttgtcagggc agcggcccgg gcaacaagag ccagctgtac
     1201 cctgacagcc ctctgaccga ccaggacttc aaccagctgg accagaccgt gattgaagcc
     1261 gccagaaggc agctcgtcgg cagaagattc atcgagctgt acggccctct cggcagaggc
     1321 atgcagagcg tgttcaacga catcttcatg gaaagccacg aggccaagat ggacttccag
     1381 ggcagcttcg acaccgaggt ggaaagcagc agaagagtga actacacaat ccccatgctg
     1441 tacaaggact tcgtgctgta ctggcgcgac ctggaacaga gcaaggccct ggacatcccc
     1501 atcgattttt ccgtggccgc caacgccgct agagatgtgg cctttctgga agatcagatg
     1561 atctttcacg gcagcaaaga gttcgacatc cccggcctga tgaacgtgaa gggcagactg
     1621 acacacctga tcggcaattg gtacgagagc ggcaacgcct tccaggacat cgtggaagcc
     1681 cggaacaagc tgctggaaat gaaccacaac ggcccctacg ctctggtgct gagccctgag
     1741 ctgtatagcc tgctgcacag agtgcacaag gacaccaacg tcctggaaat cgagcacgtg
     1801 cgcgagctga ttaccgccgg tgtctttcag agccctgtgc tgaagggaaa gtccggcgtg
     1861 atcgtgaaca ccggcagaaa caacctggac ctggccatca gcgaggactt cgagacagcc
     1921 tatctgggcg aagagggcat gaatcacccc ttccgggtgt acgagacagt ggtgctgcgg
     1981 attaagaggc ctgccgccat ctgcaccctg atcgatcctg aagaaggctc cggcgactac
     2041 aaggacgacg acgacaaatg agcggccgct cgagtctaga gggcccgttt aaacccgctg
     2101 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2161 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2221 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2281 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     2341 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     2401 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     2461 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2521 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     2581 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     2641 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     2701 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     2761 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     2821 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2881 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2941 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3001 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3061 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     3121 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     3181 ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat
     3241 aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca
     3301 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg
     3361 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
     3421 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg
     3481 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc
     3541 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg
     3601 ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg
     3661 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     3721 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     3781 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3841 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     3901 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3961 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4021 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4081 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4141 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4201 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4261 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4321 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4381 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4441 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4501 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4561 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4621 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4681 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4741 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4801 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4861 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4921 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4981 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5041 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5101 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5161 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5221 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5281 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5341 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5401 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5461 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5521 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5581 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5641 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5701 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5761 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5821 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5881 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5941 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6001 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6061 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
