LOCUS       sequence-393233-        4731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: C1 Cloning Vector, Excitation: 516, Emission: 529.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4731)
  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 4731)
  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-C1
FEATURES             Location/Qualifiers
     source          1..4731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             11..805
                     /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             11..805
                     /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(65..84)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     675..694
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(993..1012)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1037..1084
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1413..2001
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1902..1921
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2176..2479
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2480..2683
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2634..2654
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2722..2731
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2728..2730,2731..2733,2734..3444)
                     /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(2773..2794)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3034..3053)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3381..3402
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3445..3510
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3634..3755
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3671..3690)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3725..3744
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3762..4217)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3899..3920)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4111..4130
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4244..4348
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4350..4707
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4350..4541
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4373..4393)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4558..4693
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
       61 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
      121 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
      181 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
      241 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
      301 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
      361 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
      421 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
      481 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
      541 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
      601 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
      661 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
      721 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
      781 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
      841 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
      901 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
      961 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     1021 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     1081 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     1141 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     1201 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     1261 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     1321 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     1381 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     1441 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     1501 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     1561 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     1621 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     1681 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     1741 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     1801 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     1861 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     1921 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     1981 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2041 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     2101 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc
     2161 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     2221 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     2281 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     2341 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     2401 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     2461 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     2521 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     2581 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     2641 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
     2701 gatccgctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     2761 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     2821 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     2881 ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cctgatgtgc
     2941 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     3001 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     3061 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     3121 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     3181 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     3241 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     3301 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac
     3361 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     3421 ctcggcatgg acgagctgta caagtccgga ctcagatctc gagctcaagc ttcgaattct
     3481 gcagtcgacg gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc
     3541 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     3601 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     3661 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3721 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     3781 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3841 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3901 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3961 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4021 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4081 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4141 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4201 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4261 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4321 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     4381 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4441 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4501 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4561 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4621 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4681 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag g
//
