LOCUS       sequence-492634-        6625 bp    DNA     circular SYN 12-NOV-2025
DEFINITION  Localization: Centromere, Excitation: 487, Emission: 509.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6625)
  TITLE     Davidson mEmerald
REFERENCE   2  (bases 1 to 6625)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 12, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: mEmerald-Rad21-N-18
FEATURES             Location/Qualifiers
     source          1..6625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        320..523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     474..494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2521..2530
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2527..2529,2530..2532,2533..3246)
                     /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(2572..2593)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2833..2852)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3180..3201
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3368..3489
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3405..3424)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3459..3478
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3496..3951)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3633..3654)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3845..3864
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3978..4082
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4084..4441
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4084..4275
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4107..4127)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4292..4427
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4476..5270
                     /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             4476..5270
                     /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(4530..4549)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5140..5159
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5458..5477)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5502..5549
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5878..6466
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6367..6386
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
       61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      541 gatccgctag cgctaccgga ctcagatctc gaggccacca tgttctacgc acattttgtt
      601 ctcagtaaaa gagggcctct ggccaaaatt tggctagcgg cccattggga taagaagcta
      661 accaaagccc atgtgttcga gtgtaattta gagagcagcg tggagagtat catctcacca
      721 aaggtgaaaa tggcattacg gacatcagga catctcttac tgggagtagt tcgaatctat
      781 cacaggaaag ccaaatacct tcttgcagac tgtaatgaag cattcattaa gataaagatg
      841 gcttttcggc caggtgtggt tgacctgcct gaggaaaatc gggaagcagc ttataatgcc
      901 attactttac ctgaagaatt tcatgacttt gatcagccac tgcctgactt agatgacatc
      961 gatgtggccc agcagttcag cttgaatcag agtagagtgg aagagataac catgagagaa
     1021 gaagttggga acatcagtat tttacaagaa aatgattttg gtgattttgg aatggatgat
     1081 cgtgagataa tgagagaagg cagtgctttt gaggatgacg acatgttagt aagcactact
     1141 acttctaacc tcctattaga gtctgaacag agcaccagca atctgaatga gaaaattaac
     1201 catttagaat atgaagatca atataaggat gataattttg gagaaggaaa tgatggtgga
     1261 atattagatg acaaacttat tagtaataat gatggcggta tctttgatga tccccctgcc
     1321 ctctctgagg caggggtgat gttgccagag cagcctgcac atgacgatat ggatgaggat
     1381 gataatgtat caatgggtgg gcctgatagt cctgattcag tggatcccgt tgaaccaatg
     1441 ccaaccatga ctgatcaaac aacacttgtt ccaaatgagg aagaagcatt tgcattggag
     1501 cctattgata taactgttaa agaaacaaaa gccaagagga agaggaagct aattgttgac
     1561 agtgtcaaag agttggatag caagacaatt agagcccaac ttagtgatta ttcagatatt
     1621 gttactactt tggatctggc accgcccacc aagaaattga tgatgtggaa agagacagga
     1681 ggagtagaaa aactgttttc tttacctgct cagcctttgt ggaataacag actactgaag
     1741 ctctttacac gctgtcttac accgcttgta ccagaagacc ttagaaaaag gaggaaagga
     1801 ggagaggcag ataatttgga tgaattcctc aaagaatttg aaaatccaga ggttcctaga
     1861 gaggaccagc aacagcagca tcagcagcgt gatgttatcg atgagcccat tattgaagag
     1921 ccaagccgcc tccaggagtc agtgatggag gccagcagaa caaacataga tgagtcagct
     1981 atgcctccac caccacctca gggagttaag cgaaaagctg gacaaattga cccagagcct
     2041 gtgatgcctc ctcagcaggt agagcagatg gaaataccac ctgtagagct tcccccagaa
     2101 gaacctccaa atatctgtca gctaatacca gagttagaac ttctgccaga aaaagagaag
     2161 gagaaagaga aggaaaaaga agatgatgaa gaggaagagg atgaagatgc atcagggggc
     2221 gatcaagatc aggaagaaag aagatggaac aaaaggactc agcagatgct tcatggtctt
     2281 cagcgtgctc ttgctaaaac tggagctgaa tctatcagtt tgcttgagtt atgtcgaaat
     2341 acgaacagaa aacaagctgc cgcaaagttc tacagcttct tggttcttaa aaagcagcaa
     2401 gctattgagc tgacacagga agaaccgtac agtgacatca tcgcaacacc tggaccaagg
     2461 ttccatatta taagtcgcgg aggcagtggt ggggtaccgc gggcccggga tccaccggtc
     2521 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2581 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     2641 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     2701 cccaccctcg tgaccacctt gacctacggc gtgcagtgct tcgcccgcta ccccgaccac
     2761 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     2821 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     2881 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     2941 gggcacaagc tggagtacaa ctacaacagc cacaaggtct atatcaccgc cgacaagcag
     3001 aagaacggca tcaaggtgaa cttcaagacc cgccacaaca tcgaggacgg cagcgtgcag
     3061 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3121 aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
     3181 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     3241 aagtaagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     3301 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     3361 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3421 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3481 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3541 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3601 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3661 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3721 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3781 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3841 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3901 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3961 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4021 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4081 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4141 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4201 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4261 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4321 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4381 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4441 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4501 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4561 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4621 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     4681 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4741 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4801 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4861 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4921 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4981 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     5041 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5101 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5161 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5221 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5281 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5341 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5401 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     5461 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     5521 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     5581 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     5641 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     5701 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     5761 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5821 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5881 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5941 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6001 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6061 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6121 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6181 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6241 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6301 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6361 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6421 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6481 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6541 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa
     6601 ttacggggtc attagttcat agccc
//
