LOCUS       sequence-181971-        5217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of FtnA-Ypet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5217)
  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 5217)
  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: pFtnA-Ypet
FEATURES             Location/Qualifiers
     source          1..5217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      8..596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     497..516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        771..1074
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1075..1278
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1229..1249
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1311..1320
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1317..1319,1320..1322,1323..2033)
                     /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(1623..1642)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2715..2836
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2752..2771)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2806..2825
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2843..3298)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2980..3001)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3192..3211
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3325..3429
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3431..3788
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3431..3622
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3454..3474)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3639..3774
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3823..4617
                     /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             3823..4617
                     /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(3877..3896)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4487..4506
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4805..4824)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4849..4896
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
       61 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      121 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
      181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
      421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
      481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
      541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
      601 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
      661 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
      721 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      781 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      841 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      901 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      961 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     1021 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     1081 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     1141 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     1201 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     1261 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gccaccggtc gccaccatgg
     1321 tgagcaaagg cgaagagctg ttcaccggcg tggtgcccat cctggtggag ctggacggcg
     1381 acgtgaacgg ccacaagttc agcgtgagcg gcgagggcga gggcgacgcc acctacggca
     1441 agctgaccct gaagctgctg tgcaccaccg gcaagctgcc cgtgccctgg cccaccctgg
     1501 tgaccaccct gggctacggc gtgcagtgct tcgcccggta ccccgaccac atgaagcagc
     1561 acgacttctt caagagcgcc atgcccgagg gctacgtgca ggagcggacc atcttcttca
     1621 aggacgacgg caactacaag acccgggccg aggtgaagtt cgagggcgac accctggtga
     1681 accggatcga gctgaagggc atcgacttca aggaggacgg caacatcctg ggccacaagc
     1741 tggagtacaa ctacaacagc cacaacgtgt acatcaccgc cgacaagcag aagaacggca
     1801 tcaaggccaa cttcaagatc cggcacaaca tcgaggacgg cggcgtgcag ctggccgacc
     1861 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1921 tgagctacca gagcaagctg ttcaaggacc ccaacgagaa gcgggaccac atggtgctgc
     1981 tggagttcct gaccgccgcc ggcatcaccg agggcatgaa cgagctctat aaggctagcg
     2041 gagcatccgc aggaatgctg aaaccagaaa tgattgaaaa acttaatgag cagatgaacc
     2101 tggaactgta ctcttcactg ctttatcagc aaatgagcgc ctggtgcagc tatcatacct
     2161 tcgaaggtgc tgccgcgttc ctgcgccgtc acgcccagga agagatgacg catatgcagc
     2221 gtctgtttga ttacctgact gataccggca atttaccgcg tattaatacc gttgaatctc
     2281 cgtttgctga atattcctca cttgatgaat tattccagga aacctataaa cacgaacaat
     2341 taatcaccca gaaaattaac gaactggctc atgctgcaat gaccaatcag gactacccaa
     2401 catttaattt cctgcaatgg tatgtttctg agcagcatga agaagagaaa ctgttcaaat
     2461 cgattattga taaattaagc ctggcaggca aaagcggcga aggtctgtat tttatcgaca
     2521 aagaactctc taccctcgac acacaaaact aacggccgca tcgataacac cgcaaatgct
     2581 tctagggatc caccggatct agataactga tcataatcag ccataccaca tttgtagagg
     2641 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     2701 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2761 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2821 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     2881 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     2941 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3001 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3061 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3121 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3181 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     3241 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     3301 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3361 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3421 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     3481 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3541 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3601 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3661 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3721 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3781 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     3841 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3901 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3961 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     4021 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4081 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4141 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4201 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4261 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4321 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     4381 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4441 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4501 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4561 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4621 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4681 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4741 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     4801 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     4861 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     4921 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     4981 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     5041 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     5101 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5161 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaag
//
