LOCUS       sequence-181963-        6117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of GlnA-Ypet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6117)
  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 6117)
  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: pGlnA-Ypet
FEATURES             Location/Qualifiers
     source          1..6117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      9..597
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     498..517
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        772..1075
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1076..1279
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1230..1250
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1312..1321
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1318..1320,1321..1323,1324..2034)
                     /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(1624..1643)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3616..3737
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3653..3672)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3707..3726
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3744..4199)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3881..3902)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4093..4112
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4226..4330
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4332..4689
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4332..4523
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4355..4375)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4540..4675
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4724..5518
                     /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             4724..5518
                     /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(4778..4797)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5388..5407
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5706..5725)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5750..5797
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
       61 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
      121 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
      181 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
      241 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
      301 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
      361 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
      421 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
      481 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      541 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      601 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
      661 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta
      721 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      781 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      841 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      901 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      961 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     1021 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     1081 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     1141 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     1201 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     1261 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgccaccggt cgccaccatg
     1321 gtgagcaaag gcgaagagct gttcaccggc gtggtgccca tcctggtgga gctggacggc
     1381 gacgtgaacg gccacaagtt cagcgtgagc ggcgagggcg agggcgacgc cacctacggc
     1441 aagctgaccc tgaagctgct gtgcaccacc ggcaagctgc ccgtgccctg gcccaccctg
     1501 gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccggt accccgacca catgaagcag
     1561 cacgacttct tcaagagcgc catgcccgag ggctacgtgc aggagcggac catcttcttc
     1621 aaggacgacg gcaactacaa gacccgggcc gaggtgaagt tcgagggcga caccctggtg
     1681 aaccggatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct gggccacaag
     1741 ctggagtaca actacaacag ccacaacgtg tacatcaccg ccgacaagca gaagaacggc
     1801 atcaaggcca acttcaagat ccggcacaac atcgaggacg gcggcgtgca gctggccgac
     1861 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1921 ctgagctacc agagcaagct gttcaaggac cccaacgaga agcgggacca catggtgctg
     1981 ctggagttcc tgaccgccgc cggcatcacc gagggcatga acgagctcta taagtccgca
     2041 ggaatgtccg ctgaacacgt actgacgatg ctgaacgagc acgaagtgaa gtttgttgat
     2101 ttgcgcttca ccgatactaa aggtaaagaa cagcacgtca ctatccctgc tcatcaggtg
     2161 aatgctgaat tcttcgaaga aggcaaaatg tttgacggct cctcgattgg cggctggaaa
     2221 ggcattaacg agtccgacat ggtgctgatg ccagacgcat ccaccgcagt gattgacccg
     2281 ttcttcgccg actccaccct gattatccgt tgcgacatcc ttgaacctgg caccctgcaa
     2341 ggctatgacc gtgacccgcg ctccattgcg aagcgcgccg aagattacct gcgttccact
     2401 ggcattgccg acaccgtact gttcgggcca gaacctgaat tcttcctgtt cgatgacatc
     2461 cgtttcggat catctatctc cggttcccac gttgctatcg acgatatcga aggcgcatgg
     2521 aactcctcca cccaatacga aggtggtaac aaaggtcacc gtccggcagt gaaaggcggt
     2581 tacttcccgg ttccaccggt agactcggct caggatattc gttctgaaat gtgtctggtg
     2641 atggaacaga tgggtctggt ggttgaagcc catcaccacg aagtagcgac tgctggtcag
     2701 aacgaagtgg ctacccgctt caataccatg accaaaaaag ctgacgaaat tcagatctac
     2761 aaatatgttg tgcacaacgt agcgcaccgc ttcggtaaaa ccgcgacctt tatgccaaaa
     2821 ccgatgttcg gtgataacgg ctccggtatg cactgccaca tgtctctgtc taaaaacggc
     2881 gttaacctgt tcgcaggcga caaatacgca ggtctgtctg agcaggcgct gtactacatt
     2941 ggcggcgtaa tcaaacacgc taaagcgatt aacgccctgg caaacccgac caccaactct
     3001 tataagcgtc tggtcccggg ctatgaagca ccggtaatgc tggcttactc tgcgcgtaac
     3061 cgttctgcgt ctatccgtat tccggtggtt tcttctccga aagcacgtcg tatcgaagta
     3121 cgtttcccgg atccggcagc taacccgtac ctgtgctttg ctgccctgct gatggccggt
     3181 cttgatggta tcaagaacaa gatccatccg ggcgaagcca tggacaaaaa cctgtatgac
     3241 ctgccgccag aagaagcgaa agagatccca caggttgcag gctctctgga agaagcactg
     3301 aacgaactgg atctggaccg cgagttcctg aaagccggtg gcgtgttcac tgacgaagca
     3361 attgatgcgt acatcgctct gcgtcgcgaa gaagatgacc gcgtgcgtat gactccgcat
     3421 ccggtagagt ttgagctgta ctacagcgtc taacggccgc atcgataaca ccgcaaatgc
     3481 ttctagggat ccaccggatc tagataactg atcataatca gccataccac atttgtagag
     3541 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     3601 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3661 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3721 ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     3781 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     3841 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     3901 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3961 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     4021 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     4081 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     4141 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     4201 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4261 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4321 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     4381 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4441 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4501 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4561 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4621 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4681 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4741 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4801 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4861 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     4921 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4981 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5041 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5101 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5161 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5221 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     5281 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5341 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5401 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5461 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5521 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     5581 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5641 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     5701 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     5761 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     5821 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     5881 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     5941 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     6001 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6061 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaa
//
