LOCUS       sequence-474311-        5977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Endoplasmic Reticulum, Excitation: 487, Emission: 509.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5977)
  TITLE     Davidson mEmerald
REFERENCE   2  (bases 1 to 5977)
  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-Calreticulin-N-16
FEATURES             Location/Qualifiers
     source          1..5977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1858..1867
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1864..1866,1867..1869,1870..2583)
                     /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(1909..1930)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2170..2189)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2517..2538
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2705..2826
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2742..2761)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2796..2815
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2833..3288)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2970..2991)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3182..3201
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3315..3419
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3421..3778
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3421..3612
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3444..3464)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3629..3764
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3813..4607
                     /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             3813..4607
                     /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(3867..3886)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4477..4496
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4795..4814)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4839..4886
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5215..5803
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5704..5723
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgaggc caccatgctg ctatccgtgc cgctgctgct cggcctcctc
      601 ggcctggccg tcgccgagcc tgccgtctac ttcaaggagc agtttctgga cggagacggg
      661 tggacttccc gctggatcga atccaaacac aagtcagatt ttggcaaatt cgttctcagt
      721 tccggcaagt tctacggtga cgaggagaaa gataaaggtt tgcagacaag ccaggatgca
      781 cgcttttatg ctctgtcggc cagtttcgag cctttcagca acaaaggcca gacgctggtg
      841 gtgcagttca cggtgaaaca tgagcagaac atcgactgtg ggggcggcta tgtgaagctg
      901 tttcctaata gtttggacca gacagacatg cacggagact cagaatacaa catcatgttt
      961 ggtcccgaca tctgtggccc tggcaccaag aaggttcatg tcatcttcaa ctacaagggc
     1021 aagaacgtgc tgatcaacaa ggacatccgt tgcaaggatg atgagtttac acacctgtac
     1081 acactgattg tgcggccaga caacacctat gaggtgaaga ttgacaacag ccaggtggag
     1141 tccggctcct tggaagacga ttgggacttc ctgccaccca agaagataaa ggatcctgat
     1201 gcttcaaaac cggaagactg ggatgagcgg gccaagatcg atgatcccac agactccaag
     1261 cctgaggact gggacaagcc cgagcatatc cctgaccctg atgctaagaa gcccgaggac
     1321 tgggatgaag agatggacgg agagtgggaa cccccagtga ttcagaaccc tgagtacaag
     1381 ggtgagtgga agccccggca gatcgacaac ccagattaca agggcacttg gatccaccca
     1441 gaaattgaca accccgagta ttctcccgat cccagtatct atgcctatga taactttggc
     1501 gtgctgggcc tggacctctg gcaggtcaag tctggcacca tctttgacaa cttcctcatc
     1561 accaacgatg aggcatacgc tgaggagttt ggcaacgaga cgtggggcgt aacaaaggca
     1621 gcagagaaac aaatgaagga caaacaggac gaggagcaga ggcttaagga ggaggaagaa
     1681 gacaagaaac gcaaagagga ggaggaggca gaggacaagg aggatgatga ggacaaagat
     1741 gaggatgagg aggatgagga ggacaaggag gaagatgagg aggaagatgt ccccggccag
     1801 gccaaggacg agctggcatc cgggggtgcg tcaggcggca gcggatctgg accggtcgcc
     1861 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1921 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1981 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     2041 accctcgtga ccaccttgac ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg
     2101 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2161 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2221 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2281 cacaagctgg agtacaacta caacagccac aaggtctata tcaccgccga caagcagaag
     2341 aacggcatca aggtgaactt caagacccgc cacaacatcg aggacggcag cgtgcagctc
     2401 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     2461 cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     2521 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     2581 taagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     2641 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     2701 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2761 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2821 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     2881 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     2941 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3001 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3061 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3121 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3181 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3241 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     3301 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3361 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     3421 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3481 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3541 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3601 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3661 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     3721 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     3781 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3841 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3901 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3961 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     4021 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4081 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4141 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4201 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4261 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4321 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4381 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4441 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4501 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4561 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4621 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4681 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4741 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     4801 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     4861 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     4921 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     4981 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     5041 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     5101 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5161 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5221 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5281 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5341 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5401 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5461 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5521 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5581 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5641 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5701 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5761 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5821 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5881 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
     5941 cggggtcatt agttcatagc ccatatatgg agttccg
//
