LOCUS       sequence-209109-        7508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fluorescent fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7508)
  TITLE     Johannes Schmid plasmids
REFERENCE   2  (bases 1 to 7508)
  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"
COMMENT     Sequence Label: EGFP-AR
FEATURES             Location/Qualifiers
     source          1..7508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4296..4417
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4333..4352)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4387..4406
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4424..4879)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4561..4582)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4773..4792
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4906..5010
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5012..5369
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5012..5203
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5035..5055)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5220..5355
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5404..6198
                     /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             5404..6198
                     /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(5458..5477)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6068..6087
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6386..6405)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6430..6477
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6806..7394
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7295..7314
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagga attcggtgga agctacagac aagctcaagg
     1381 atggaggtgc agttagggct gggaagggtc tacccacggc ccccatccaa gacctatcga
     1441 ggagcgttcc agaatctgtt ccagagcgtg cgcgaagcga tccagaaccc gggccccagg
     1501 caccctgagg ccgctaacat agcacctccc ggcgcctgtt tacagcagag gcaggagact
     1561 agcccccggc ggcggcggcg gcagcagcac actgaggatg gttctcctca agcccacatc
     1621 agaggcccca caggctacct ggccctggag gaggaacagc agccttcaca gcagcaggca
     1681 gcctccgagg gccaccctga gagcagctgc ctccccgagc ctggggcggc caccgctcct
     1741 ggcaaggggc tgccgcagca gccaccagct cctccagatc aggatgactc agctgcccca
     1801 tccacgttgt ccctgctggg ccccactttc ccaggcttaa gcagctgctc cgccgacatt
     1861 aaagacattt tgaacgaggc cggcaccatg caacttcttc agcagcagca acaacagcag
     1921 cagcaccaac agcagcacca acagcaccaa cagcagcagg aggtaatctc cgaaggcagc
     1981 agcgcaagag ccagggaggc cacgggggct ccctcttcct ccaaggatag ttacctaggg
     2041 ggcaattcaa ccatatctga cagtgccaag gagttgtgta aagcagtgtc tgtgtccatg
     2101 ggattgggtg tggaagcatt ggaacatctg agtccagggg aacagcttcg gggagactgc
     2161 atgtacgcgt cgctcctggg aggtccaccc gcggtgcgtc ccactccttg tgcgccgctg
     2221 cccgaatgca aaggtcttcc cctggacgaa ggcccaggca aaagcactga agagactgct
     2281 gagtattcct ctttcaaggg aggttacgcc aaaggattgg aaggtgagag cttggggtgc
     2341 tctggcagca gtgaagcagg tagctctggg acacttgaga tcccgtcctc tctgtctctg
     2401 tataaatctg gagcactaga cgaggcagca gcataccaga atcgcgacta ctacaacttt
     2461 ccgctggctc tgtccgggcc gccgcacccc ccgcccccta cccatccaca cgcccgtatc
     2521 aagctggaga acccattgga ctacggcagc gcctgggctg cggcggcagc gcaatgccgc
     2581 tatggggact tgggtagtct acatggaggg agtgtagccg ggcccagcac tggatcgccc
     2641 ccagccacca cctcttcttc ctggcatact ctcttcacag ctgaagaagg ccaattatat
     2701 gggccaggag gcgggggcgg cagcagcagc ccaagcgatg ccgggcctgt agccccctat
     2761 ggctacactc ggccccctca ggggctgaca agccaggaga gtgactactc tgcctccgaa
     2821 gtgtggtatc ctggtggagt tgtgaacaga gtaccctatc ccagtcccaa ttgtgtcaaa
     2881 agtgaaatgg gaccttggat ggagaactac tccggacctt atggggacat gcgtttggac
     2941 agtaccaggg accatgtttt acccatcgac tattactttc caccccagaa gacctgcctg
     3001 atctgtggag atgaagcttc tggctgtcac tacggagctc tcacttgtgg cagctgcaag
     3061 gtcttcttca aaagagccgc tgaagggaaa cagaagtatc tatgtgccag cagaaacgat
     3121 tgtaccattg ataaatttcg gaggaaaaat tgcccatctt gtcgtctccg gaaatgttat
     3181 gaagcaggga tgactctggg agctcgtaag ctgaagaaac ttggaaatct aaaactacag
     3241 gaggaaggag aaaactccaa tgctggcagc cccactgagg acccatccca gaagatgact
     3301 gtatcacaca ttgaaggcta tgaatgtcag cctatctttc ttaacgtcct ggaagccatt
     3361 gagccaggag tggtgtgtgc cggacatgac aacaaccaac cagattcctt tgctgccttg
     3421 ttatctagcc tcaatgagct tggagagagg cagcttgtgc atgtggtcaa gtgggccaag
     3481 gccttgcctg gcttccgcaa cttgcatgtg gatgaccaga tggcggtcat tcagtattcc
     3541 tggatgggac tgatggtatt tgccatgggt tggcggtcct tcactaatgt caactccagg
     3601 atgctctact ttgcacctga cttggttttc aatgagtacc gcatgcacaa gtctcggatg
     3661 tacagccagt gtgtgaggat gaggcacctg tctcaagagt ttggatggct ccaaataacc
     3721 ccccaggaat tcctgtgcat gaaagcactg ctgctcttca gcattattcc agtggatggg
     3781 ctgaaaaatc aaaaattctt tgatgaactt cgaatgaact acatcaagga actcgatcgc
     3841 atcattgcat gcaaaagaaa gaatcccaca tcctgctcaa ggcgcttcta ccagctcacc
     3901 aagctcctgg attctgtgca gcctattgca agagagctgc atcagttcac ttttgacctg
     3961 ctaatcaagt cccatatggt gagcgtggac tttcctgaaa tgatggcaga gatcatctct
     4021 gtgcaagtgc ccaagatcct ttctgggaaa gtcaagccca tctatttcca cacacagtgg
     4081 agatttggaa accctaatac ccaaaaccca ccttgttccc tttccagatg tcttctgcct
     4141 gttatataac tctgcactac ttctctggat ccaccggatc tagataactg atcataatca
     4201 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     4261 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     4321 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4381 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     4441 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     4501 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     4561 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4621 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     4681 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     4741 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     4801 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     4861 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4921 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4981 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     5041 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5101 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5161 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     5221 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     5281 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     5341 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     5401 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5461 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5521 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5581 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5641 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5701 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5761 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5821 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5881 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     5941 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     6001 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     6061 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     6121 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     6181 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     6241 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     6301 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     6361 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     6421 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     6481 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     6541 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     6601 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     6661 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6721 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6781 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6841 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6901 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     6961 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7021 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7081 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7141 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7201 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7261 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7321 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7381 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7441 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     7501 ccatgcat
//
