LOCUS       sequence-358017-        4732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: N1 Cloning Vector, Excitation: 516, Emission: 529.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4732)
  AUTHORS   Griesbeck O, Baird GS, Campbell RE, Zacharias DA, Tsien RY
  TITLE     Reducing the environmental sensitivity of yellow fluorescent 
            protein. Mechanism and applications.
  JOURNAL   J Biol Chem. 2001 Aug 3;276(31):29188-94. Epub 2001 May 31.
  PUBMED    11387331
REFERENCE   2  (bases 1 to 4732)
  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: "J Biol 
            Chem."; date: "2001-08-3"; volume: "276(31)"; pages: "29188-94. Epub
            2001 May 31"
COMMENT     Sequence Label: mCitrine-N1
FEATURES             Location/Qualifiers
     source          1..4732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6..141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        158..349
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     306..326
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(351..455)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      482..937
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(569..588)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     779..800
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    944..1065
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(955..974)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1009..1028
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1187..1900,1901..1903,1904..1906))
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1232..1253)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1581..1600
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1840..1861
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1903..1912
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1914..1994
                     /label=MCS
                     /note="multiple cloning site"
     promoter        complement(2017..2220)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2046..2066)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2221..2524
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2699..3287)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2779..2798)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3616..3663
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3688..3707
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3895..4689)
                     /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             complement(3895..4689)
                     /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(4006..4025)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4616..4635
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(4724..4732,1..349))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
       61 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
      121 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
      181 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
      241 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
      301 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcagg actcttcctt
      361 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
      421 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
      481 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
      541 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      601 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      661 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      721 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      781 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      841 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
      901 aacgcgaatt ttaacaaaat attaacgctt acaatttacg ccttaagata cattgatgag
      961 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     1021 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     1081 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     1141 ctctacaaat gtggtatggc tgattatgat ctagagtcgc ggccgcttac ttgtacagct
     1201 cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc
     1261 gcttctcgtt ggggtctttg ctcagcttgg actggtagct caggtagtgg ttgtcgggca
     1321 gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc
     1381 tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc ttctgcttgt
     1441 cggccatgat atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt
     1501 tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct
     1561 cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct
     1621 cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg
     1681 ggtagcgggc gaagcacatc aggccgtagc cgaaggtggt cacgagggtg ggccagggca
     1741 cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc
     1801 cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca
     1861 ggatgggcac caccccggtg aacagctcct cgcccttgct caccatggtg gcgaccggtg
     1921 gatcccgggc ccgcggtacc gtcgactgca gaattcgaag cttgagctcg agatctgagt
     1981 ccggtagcgc tagcggatct gacggttcac taaaccagct ctgcttatat agacctccca
     2041 ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa
     2101 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     2161 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac
     2221 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
     2281 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
     2341 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     2401 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     2461 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     2521 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata
     2581 actaatgcat ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2641 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2701 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2761 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2821 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2881 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2941 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3001 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3061 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3121 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3181 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3241 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3301 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3361 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3421 caatctaaag tatatatgag taacctgagg ctatggcagg gcctgccgcc ccgacgttgg
     3481 ctgcgagccc tgggccttca cccgaacttg gggggtgggg tggggaaaag gaagaaacgc
     3541 gggcgtattg gccccaatgg ggtctcggtg gggtatcgac agagtgccag ccctgggacc
     3601 gaaccccgcg tttatgaaca aacgacccaa caccgtgcgt tttattctgt ctttttattg
     3661 ccgtcatagc gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc
     3721 tagggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt
     3781 cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc
     3841 gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa
     3901 gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta
     3961 aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc
     4021 caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga
     4081 aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag
     4141 atcctcgccg tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc
     4201 ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc
     4261 tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg
     4321 cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga
     4381 caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac
     4441 aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc
     4501 ctcgtcttgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg
     4561 cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca
     4621 gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg
     4681 ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa aa
//
