LOCUS       sequence-181964-        5379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of Ypet-FsaA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5379)
  AUTHORS   Finan K, Raulf A, Heilemann M
  TITLE     A set of homo-oligomeric standards allows accurate protein counting.
  JOURNAL   Angew Chem Int Ed Engl. 2015 Oct 5;54(41):12049-52. doi: 
            10.1002/anie.201505664. Epub 2015 Aug 20.
  PUBMED    26289028
REFERENCE   2  (bases 1 to 5379)
  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"; doi: 
            "10.1002/anie.201505664"; journalName: "Angew Chem Int Ed Engl"; 
            date: "2015-10-5- 5"; volume: "54"; issue: "41"; pages: "12049-52"
COMMENT     Sequence Label: pYpet-FsaA
FEATURES             Location/Qualifiers
     source          1..5379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      3..591
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     492..511
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        766..1069
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1070..1273
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1224..1244
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1306..1315
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1990..1992,1993..1995,1996..2709)
                     /codon_start=1
                     /product="yellow fluorescent protein optimized for FRET 
                     with CyPet (Nguyen and Daugherty, 2005)"
                     /label=YPet
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLLCTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLFKDPNEKRDHMVLL
                     EFLTAAGITEGMNELYK"
     primer_bind     complement(2296..2315)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2872..2993
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2909..2928)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2963..2982
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3000..3455)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3137..3158)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3349..3368
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3482..3586
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3588..3945
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3588..3779
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3611..3631)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3796..3931
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3980..4774
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3980..4774
                     /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(4034..4053)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4644..4663
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4962..4981)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5006..5053
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
       61 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      121 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
      181 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      241 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      301 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      361 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      421 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      481 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      541 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      601 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      661 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
      721 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
      781 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      841 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      901 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      961 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     1021 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     1081 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     1141 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     1201 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     1261 tataagcaga gctggtttag tgaaccgtca gatccgccac cggtcgccac catggaactg
     1321 tatctggata cttcagacgt tgttgcggtg aaggcgctgt cacgtatttt tccgctggcg
     1381 ggtgtgacca ctaacccaag cattatcgcc gcgggtaaaa aaccgctgga tgttgtgctt
     1441 ccgcaacttc atgaagcgat gggcggtcag gggcgtctgt ttgcccaggt aatggctacc
     1501 actgccgaag ggatggttaa tgacgcgctt aagctgcgtt ctattattgc ggatatcgtg
     1561 gtgaaagttc cggtgaccgc cgaggggctg gcagctatta agatgttaaa agcggaaggg
     1621 attccgacgc tgggaaccgc ggtatatggc gcagcacaag ggctgctgtc ggcgctggca
     1681 ggtgcggaat atgttgcgcc ttacgttaat cgtattgatg ctcagggcgg tagcggcatt
     1741 cagactgtga ccgacttaca ccagttattg aaaatgcatg cgccgcaggc gaaagtgctg
     1801 gcagcgagtt tcaaaacccc gcgtcaggcg ctggactgct tactggcagg atgtgaatca
     1861 attactctgc cactggatgt ggcacaacag atgattagct atccggcggt tgatgccgct
     1921 gtggcgaagt ttgagcagga ctggcaggga gcgtttggca gaacgtcgat tgccggagca
     1981 tccgcaggaa tggtgagcaa aggcgaagag ctgttcaccg gcgtggtgcc catcctggtg
     2041 gagctggacg gcgacgtgaa cggccacaag ttcagcgtga gcggcgaggg cgagggcgac
     2101 gccacctacg gcaagctgac cctgaagctg ctgtgcacca ccggcaagct gcccgtgccc
     2161 tggcccaccc tggtgaccac cctgggctac ggcgtgcagt gcttcgcccg gtaccccgac
     2221 cacatgaagc agcacgactt cttcaagagc gccatgcccg agggctacgt gcaggagcgg
     2281 accatcttct tcaaggacga cggcaactac aagacccggg ccgaggtgaa gttcgagggc
     2341 gacaccctgg tgaaccggat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2401 ctgggccaca agctggagta caactacaac agccacaacg tgtacatcac cgccgacaag
     2461 cagaagaacg gcatcaaggc caacttcaag atccggcaca acatcgagga cggcggcgtg
     2521 cagctggccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2581 gacaaccact acctgagcta ccagagcaag ctgttcaagg accccaacga gaagcgggac
     2641 cacatggtgc tgctggagtt cctgaccgcc gccggcatca ccgagggcat gaacgagctc
     2701 tataagtagc ggccgcatcg ataacaccgc aaatgcttct agggatccac cggatctaga
     2761 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2821 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2881 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2941 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
     3001 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3061 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3121 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3181 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3241 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3301 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3361 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3421 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     3481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     3601 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3661 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3721 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3781 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3841 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3901 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3961 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4021 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4081 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4141 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4201 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4261 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4321 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4381 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4441 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4501 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4561 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4621 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4681 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4741 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4801 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4861 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4921 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4981 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     5041 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     5101 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     5161 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     5221 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5281 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5341 cactgagcgt cagaccccgt agaaaagatc aaaggatct
//
