LOCUS       sequence-311823-        6045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Microtubules, Excitation: 488, Emission: 507.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6045)
  AUTHORS   Rizzo MA, Davidson MW, Piston DW
  TITLE     Fluorescent protein tracking and detection: fluorescent protein 
            structure and color variants.
  JOURNAL   Cold Spring Harb Protoc. 2009 Dec;2009(12):pdb.top63. doi: 
            10.1101/pdb.top63.
  PUBMED    20150100
REFERENCE   2  (bases 1 to 6045)
  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: "Cold Spring
            Harb Protoc."; date: "2009-12"; volume: "2009(12):pdb.top63. doi"; 
            pages: " 10.1101/pdb.top63"
COMMENT     Sequence Label: EGFP-Tubulin-6
FEATURES             Location/Qualifiers
     source          1..6045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        50..353
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        354..557
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     508..528
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      596..605
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(602..604,605..607,608..1318)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(602..604,605..607,608..1318)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(647..668)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(908..927)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1255..1276
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2822..2943
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2859..2878)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2913..2932
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2950..3405)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3087..3108)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3299..3318
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3432..3536
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3538..3895
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3538..3729
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3561..3581)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3746..3881
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3930..4724
                     /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             3930..4724
                     /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(3984..4003)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4594..4613
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4912..4931)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4956..5003
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5332..5920
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5821..5840
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
       61 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      121 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      181 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      241 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
      301 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      361 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      421 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      481 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
      541 tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac cggtcgccac
      601 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      661 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
      721 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
      781 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
      841 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
      901 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
      961 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1021 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     1081 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     1141 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1201 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     1261 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc
     1321 cggactcaga tctcgagtgc gtgagtgcat ctccatccac gttggccagg ctggtgtcca
     1381 gattggcaat gcctgctggg agctctactg cctggaacac ggcatccagc ccgatggcca
     1441 gatgccaagt gacaagacca ttgggggagg agatgactcc ttcaacacct tcttcagtga
     1501 gacgggcgct ggcaagcacg tgccccgggc tgtgtttgta gacttggaac ccacagtcat
     1561 tgatgaagtt cgcactggca cctaccgcca gctcttccac cctgagcagc tcatcacagg
     1621 caaggaagat gctgccaata actatgcccg agggcactac accattggca aggagatcat
     1681 tgaccttgtg ttggaccgaa ttcgcaagct ggctgaccag tgcaccggtc ttcagggctt
     1741 cttggttttc cacagctttg gtgggggaac tggttctggg ttcacctccc tgctcatgga
     1801 acgtctctca gttgattatg gcaagaagtc caagctggag ttctccattt acccagcacc
     1861 ccaggtttcc acagctgtag ttgagcccta caactccatc ctcaccaccc acaccaccct
     1921 ggagcactct gattgtgcct tcatggtaga caatgaggcc atctatgaca tctgtcgtag
     1981 aaacctcgat atcgagcgcc caacctacac taaccttaac cgccttatta gccagattgt
     2041 gtcctccatc actgcttccc tgagatttga tggagccctg aatgttgacc tgacagaatt
     2101 ccagaccaac ctggtgccct acccccgcat ccacttccct ctggccacat atgcccctgt
     2161 catctctgct gagaaagcct accatgaaca gctttctgta gcagagatca ccaatgcttg
     2221 ctttgagcca gccaaccaga tggtgaaatg tgaccctcgc catggtaaat acatggcttg
     2281 ctgcctgttg taccgtggtg acgtggttcc caaagatgtc aatgctgcca ttgccaccat
     2341 caaaaccaag cgcagcatcc agtttgtgga ttggtgcccc actggcttca aggttggcat
     2401 caactaccag cctcccactg tggtgcctgg tggagacctg gccaaggtac agagagctgt
     2461 gtgcatgctg agcaacacca cagccattgc tgaggcctgg gctcgcctgg accacaagtt
     2521 tgacctgatg tatgccaagc gtgcctttgt tcactggtac gtgggtgagg ggatggagga
     2581 aggcgagttt tcagaggccc gtgaagatat ggctgccctt gagaaggatt atgaggaggt
     2641 tggtgtggat tctgttgaag gagagggtga ggaagaagga gaggaatact aaggatccac
     2701 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2761 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2821 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2881 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2941 ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     3001 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     3061 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     3121 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     3181 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     3241 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3301 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3361 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     3421 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3481 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     3541 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3601 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3661 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3721 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3781 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3841 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     3901 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3961 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4021 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4081 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     4141 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4201 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4261 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4321 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4381 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4441 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4501 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4561 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4621 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4681 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4741 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4801 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4861 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     4921 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     4981 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     5041 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     5101 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     5161 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     5221 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5281 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5341 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5401 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5461 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5521 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5581 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5641 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5701 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5761 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5821 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5881 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5941 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6001 tgattctgtg gataaccgta ttaccgccat gcattagtta ttaat
//
