LOCUS       sequence-264404-        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        43..234
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     191..211
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(236..340)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      367..822
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(454..473)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     664..685
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    829..950
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(840..859)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     894..913
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(3917..4627,4628..4630,4631..4633))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(3917..4627,4628..4630,4631..4633))
                     /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(3959..3980)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4308..4327
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4567..4588
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4630..4639
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4678..4881)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4707..4727)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4882..5185
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5360..5948)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5440..5459)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6277..6324
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     6349..6368
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(6556..7350)
                     /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(6556..7350)
                     /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(6667..6686)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7277..7296
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(7385..7508,1..234))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7399..7508,1..26)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
       61 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
      121 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
      181 gactttccac accctaactg acacacattc cacagctggt tctttccgcc tcaggactct
      241 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
      301 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
      361 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
      421 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      481 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      541 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      601 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      661 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      721 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      781 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttacgcgtta agatacattg
      841 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
      901 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
      961 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1021 aaaacctcta caaatgtggt atggctgatt atgatcagtt atctagatcc ggtggatcca
     1081 gagaagtagt gcagagttat ataacaggca gaagacatct ggaaagggaa caaggtgggt
     1141 tttgggtatt agggtttcca aatctccact gtgtgtggaa atagatgggc ttgactttcc
     1201 cagaaaggat cttgggcact tgcacagaga tgatctctgc catcatttca ggaaagtcca
     1261 cgctcaccat atgggacttg attagcaggt caaaagtgaa ctgatgcagc tctcttgcaa
     1321 taggctgcac agaatccagg agcttggtga gctggtagaa gcgccttgag caggatgtgg
     1381 gattctttct tttgcatgca atgatgcgat cgagttcctt gatgtagttc attcgaagtt
     1441 catcaaagaa tttttgattt ttcagcccat ccactggaat aatgctgaag agcagcagtg
     1501 ctttcatgca caggaattcc tggggggtta tttggagcca tccaaactct tgagacaggt
     1561 gcctcatcct cacacactgg ctgtacatcc gagacttgtg catgcggtac tcattgaaaa
     1621 ccaagtcagg tgcaaagtag agcatcctgg agttgacatt agtgaaggac cgccaaccca
     1681 tggcaaatac catcagtccc atccaggaat actgaatgac cgccatctgg tcatccacat
     1741 gcaagttgcg gaagccaggc aaggccttgg cccacttgac cacatgcaca agctgcctct
     1801 ctccaagctc attgaggcta gataacaagg cagcaaagga atctggttgg ttgttgtcat
     1861 gtccggcaca caccactcct ggctcaatgg cttccaggac gttaagaaag ataggctgac
     1921 attcatagcc ttcaatgtgt gatacagtca tcttctggga tgggtcctca gtggggctgc
     1981 cagcattgga gttttctcct tcctcctgta gttttagatt tccaagtttc ttcagcttac
     2041 gagctcccag agtcatccct gcttcataac atttccggag acgacaagat gggcaatttt
     2101 tcctccgaaa tttatcaatg gtacaatcgt ttctgctggc acatagatac ttctgtttcc
     2161 cttcagcggc tcttttgaag aagaccttgc agctgccaca agtgagagct ccgtagtgac
     2221 agccagaagc ttcatctcca cagatcaggc aggtcttctg gggtggaaag taatagtcga
     2281 tgggtaaaac atggtccctg gtactgtcca aacgcatgtc cccataaggt ccggagtagt
     2341 tctccatcca aggtcccatt tcacttttga cacaattggg actgggatag ggtactctgt
     2401 tcacaactcc accaggatac cacacttcgg aggcagagta gtcactctcc tggcttgtca
     2461 gcccctgagg gggccgagtg tagccatagg gggctacagg cccggcatcg cttgggctgc
     2521 tgctgccgcc cccgcctcct ggcccatata attggccttc ttcagctgtg aagagagtat
     2581 gccaggaaga agaggtggtg gctgggggcg atccagtgct gggcccggct acactccctc
     2641 catgtagact acccaagtcc ccatagcggc attgcgctgc cgccgcagcc caggcgctgc
     2701 cgtagtccaa tgggttctcc agcttgatac gggcgtgtgg atgggtaggg ggcggggggt
     2761 gcggcggccc ggacagagcc agcggaaagt tgtagtagtc gcgattctgg tatgctgctg
     2821 cctcgtctag tgctccagat ttatacagag acagagagga cgggatctca agtgtcccag
     2881 agctacctgc ttcactgctg ccagagcacc ccaagctctc accttccaat cctttggcgt
     2941 aacctccctt gaaagaggaa tactcagcag tctcttcagt gcttttgcct gggccttcgt
     3001 ccaggggaag acctttgcat tcgggcagcg gcgcacaagg agtgggacgc accgcgggtg
     3061 gacctcccag gagcgacgcg tacatgcagt ctccccgaag ctgttcccct ggactcagat
     3121 gttccaatgc ttccacaccc aatcccatgg acacagacac tgctttacac aactccttgg
     3181 cactgtcaga tatggttgaa ttgcccccta ggtaactatc cttggaggaa gagggagccc
     3241 ccgtggcctc cctggctctt gcgctgctgc cttcggagat tacctcctgc tgctgttggt
     3301 gctgttggtg ctgctgttgg tgctgctgct gttgttgctg ctgctgaaga agttgcatgg
     3361 tgccggcctc gttcaaaatg tctttaatgt cggcggagca gctgcttaag cctgggaaag
     3421 tggggcccag cagggacaac gtggatgggg cagctgagtc atcctgatct ggaggagctg
     3481 gtggctgctg cggcagcccc ttgccaggag cggtggccgc cccaggctcg gggaggcagc
     3541 tgctctcagg gtggccctcg gaggctgcct gctgctgtga aggctgctgt tcctcctcca
     3601 gggccaggta gcctgtgggg cctctgatgt gggcttgagg agaaccatcc tcagtgtgct
     3661 gctgccgccg ccgccgccgg gggctagtct cctgcctctg ctgtaaacag gcgccgggag
     3721 gtgctatgtt agcggcctca gggtgcctgg ggcccgggtt ctggatcgct tcgcgcacgc
     3781 tctggaacag attctggaac gctcctcgat aggtcttgga tgggggccgt gggtagaccc
     3841 ttcccagccc taactgcacc tccatccttg agcttgtctg tagcttccac cgaattcctc
     3901 gagatctgag tccggacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     3961 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact
     4021 gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     4081 gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt
     4141 tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt
     4201 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     4261 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
     4321 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     4381 agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca
     4441 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca
     4501 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
     4561 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     4621 ccttgctcac catggtggcg accggtagcg ctagcggatc tgacggttca ctaaaccagc
     4681 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga
     4741 gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     4801 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     4861 gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca
     4921 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     4981 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     5041 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     5101 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     5161 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     5221 cgtaattgat tactattaat aactaatgca tggcggtaat acggttatcc acagaatcag
     5281 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5341 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5401 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5461 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5521 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5581 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5641 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5701 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5761 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5821 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5881 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5941 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6001 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6061 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaacctgag gctatggcag
     6121 ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc acccgaactt ggggggtggg
     6181 gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt ggggtatcga
     6241 cagagtgcca gccctgggac cgaaccccgc gtttatgaac aaacgaccca acaccgtgcg
     6301 ttttattctg tctttttatt gccgtcatag cgcgggttcc ttccggtatt gtctccttcc
     6361 gtgtttcagt tagcctcccc ctagggtggg cgaagaactc cagcatgaga tccccgcgct
     6421 ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg
     6481 cggcggtgga atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga
     6541 accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga
     6601 atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc
     6661 ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg
     6721 gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc
     6781 atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa
     6841 cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc
     6901 ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca
     6961 ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc
     7021 ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca
     7081 gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc
     7141 cagccacgat agccgcgctg cctcgtcttg cagttcattc agggcaccgg acaggtcggt
     7201 cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca
     7261 gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga
     7321 acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc
     7381 gatctttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag
     7441 aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag
     7501 aatgggcg
//
