LOCUS       sequence-298515-        6397 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Cytoskeleton, Excitation: 487, Emission: 509.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6397)
  TITLE     Davidson mEmerald
REFERENCE   2  (bases 1 to 6397)
  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: mEmerald-WASP1-N-14
FEATURES             Location/Qualifiers
     source          1..6397
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        72..375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        376..579
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     530..550
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2349..2358
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2355..2357,2358..2360,2361..3074)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2400..2421)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2661..2680)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3008..3029
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3196..3317
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3233..3252)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3287..3306
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3324..3779)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3461..3482)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3673..3692
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3806..3910
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3912..4269
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3912..4103
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3935..3955)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4120..4255
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4304..5098
                     /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             4304..5098
                     /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(4358..4377)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4968..4987
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5286..5305)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5330..5377
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5706..6294
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6195..6214
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
       61 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      121 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      181 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      241 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      301 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      361 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      421 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      481 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      541 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      601 cgctagcgct accggactca gatctcgaga ccgccatgcc gctagtgaaa agaaacatcg
      661 atcctaggca cttgtgccac acagcactgc ctagaggcat taagaatgaa ctggaatgtg
      721 taaccaatat ttccttggca aatataatta gacaactaag tagcctaagt aaatatgctg
      781 aagatatatt tggagaatta ttcaatgaag cacatagttt ttccttcaga gtcaactcat
      841 tgcaagaacg tgtggaccgt ttatctgtta gtgttacaca gcttgatcca aaggaagaag
      901 aattgtcttt gcaagatata acaatgagga aagctttccg aagttctaca attcaagacc
      961 agcagctttt cgatcgcaag actttgccta ttccattaca ggagacgtac gatgtttgtg
     1021 aacagcctcc acctctcaat atactcactc cttatagaga tgatggtaaa gaaggtctga
     1081 agttttatac caatccttcg tatttctttg atctatggaa agaaaaaatg ttgcaagata
     1141 cagaggataa gaggaaggaa aagaggaagc agaagcagaa aaatctagat cgtcctcatg
     1201 aaccagaaaa agtgccaaga gcacctcatg acaggcggcg agaatggcag aagctggccc
     1261 aaggtccaga gctggctgaa gatgatgcta atctcttaca taagcatatt gaagttgcta
     1321 atggcccagc ctctcatttt gaaacaagac ctcagacata cgtggatcat atggatggat
     1381 cttactcact ttctgccttg ccatttagtc agatgagtga gcttctgact agagctgagg
     1441 aaagggtatt agtcagacca catgaaccac ctccacctcc accaatgcat ggagcaggag
     1501 atgcaaaacc gatacccacc tgtatcagtt ctgctacagg tttgatagaa aatcgccctc
     1561 agtcaccagc tacaggcaga acacctgtgt ttgtgagccc cactccccca cctcctccac
     1621 cacctcttcc atctgccttg tcaacttcct cattaagagc ttcaatgact tcaactcctc
     1681 cccctccagt acctccccca cctccacctc cagccactgc tttgcaagct ccagcagtac
     1741 caccacctcc agctcctctt cagattgccc ctggagttct tcacccagct cctcctccaa
     1801 ttgcacctcc tctagtacag ccctctccac cagtagctag agctgcccca gtatgtgaga
     1861 ctgtaccagt tcatccactc ccacaaggtg aagttcaggg gctgcctcca cccccaccac
     1921 cgcctcctct gcctccacct ggcattcgac catcatcacc tgtcacagtt acagctcttg
     1981 ctcatcctcc ctctgggcta catccaactc catctactgc cccaggtccc catgttccat
     2041 taatgcctcc atctcctcca tcacaagtta tacctgcttc tgagccaaag cgccatccat
     2101 caaccctacc tgtaatcagt gatgccagga gtgtgctact ggaagcaata cgaaaaggta
     2161 ttcagctacg caaagtagaa gagcagcgtg aacaggaagc taagcatgaa cgcattgaaa
     2221 acgatgttgc caccatcctg tctcgccgta ttgctgttga atatagtgat tcggaagatg
     2281 attcagaatt tgatgaagta gattggttgg aggggggtgc gtcaggcggc agcggatctg
     2341 gaccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     2401 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     2461 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     2521 tgccctggcc caccctcgtg accaccttga cctacggcgt gcagtgcttc gcccgctacc
     2581 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2641 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2701 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2761 acatcctggg gcacaagctg gagtacaact acaacagcca caaggtctat atcaccgccg
     2821 acaagcagaa gaacggcatc aaggtgaact tcaagacccg ccacaacatc gaggacggca
     2881 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2941 tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc
     3001 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     3061 agctgtacaa gtaagcggcc gcgactctag atcataatca gccataccac atttgtagag
     3121 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     3181 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3241 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3301 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     3361 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     3421 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     3481 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3541 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3601 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3661 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3721 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3781 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3841 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3901 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3961 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4021 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4081 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4141 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4201 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4261 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4321 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4381 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4441 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     4501 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4561 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4621 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4681 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4741 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4801 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     4861 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4921 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4981 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5041 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5101 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     5161 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5221 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     5281 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     5341 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     5401 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     5461 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     5521 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     5581 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5641 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5701 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5761 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5821 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5881 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5941 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6001 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6061 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6121 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6181 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6241 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6301 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6361 cctgcgttat cccctgattc tgtggataac cgtatta
//
