LOCUS       sequence-243987-        6456 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Localization: Focal Adhesions/Membrane, Excitation: 487, Emission: 
            509.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6456)
  TITLE     Davidson mEmerald
REFERENCE   2  (bases 1 to 6456)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: mEmerald-Ezrin-N-14
FEATURES             Location/Qualifiers
     source          1..6456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        84..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        388..591
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     542..562
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2420..2429
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2426..2428,2429..2431,2432..3145)
                     /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(2471..2492)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2732..2751)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3079..3100
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3267..3388
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3304..3323)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3358..3377
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3395..3850)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3532..3553)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3744..3763
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3877..3981
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3983..4340
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3983..4174
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4006..4026)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4191..4326
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4375..5169
                     /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             4375..5169
                     /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(4429..4448)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5039..5058
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5357..5376)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5401..5448
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5777..6365
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6266..6285
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      421 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      481 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      541 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
      601 aaccgtcaga tccgctagcg ccaccatgcc gaaaccaatc aatgtccgag ttaccaccat
      661 ggatgcagag ctggagtttg caatccagcc aaatacaact ggaaaacagc tttttgatca
      721 ggtggtaaag actatcggcc tccgggaagt gtggtacttt ggcctccact atgtggataa
      781 taaaggattt cctacctggc tgaagctgga taagaaggtg tctgcccagg aggtcaggaa
      841 ggagaatccc ctccagttca agttccgggc caagttctac cctgaagatg tggctgagga
      901 gctcatccag gacatcaccc agaaactttt cttcctccaa gtgaaggaag gaatccttag
      961 cgatgagatc tactgccccc ctgagactgc cgtgctcttg gggtcctacg ctgtgcaggc
     1021 caagtttggg gactacaaca aagaagtgca caagtctggg tacctcagct ctgagcggct
     1081 gatccctcaa agagtgatgg accagcacaa acttaccagg gaccagtggg aggaccggat
     1141 ccaggtgtgg catgcggaac accgtgggat gctcaaagat aatgctatgt tggaatacct
     1201 gaagattgct caggacctgg aaatgtatgg aatcaactat ttcgagataa aaaacaagaa
     1261 aggaacagac ctttggcttg gagttgatgc ccttggactg aatatttatg agaaagatga
     1321 taagttaacc ccaaagattg gctttccttg gagtgaaatc aggaacatct ctttcaatga
     1381 caaaaagttt gtcattaaac ccatcgacaa gaaggcacct gactttgtgt tttatgcccc
     1441 acgtctgaga atcaacaagc ggatcctgca gctctgcatg ggcaaccatg agttgtatat
     1501 gcgccgcagg aagcctgaca ccatcgaggt gcagcagatg aaggcccagg cccgggagga
     1561 gaagcatcag aagcagctgg agcggcaaca gctggaaaca gagaagaaaa ggagagaaac
     1621 cgtggagaga gagaaagagc agatgatgcg cgagaaggag gagttgatgc tgcggctgca
     1681 ggactatgag gagaagacaa agaaggcaga gagagagctc tcggagcaga ttcagagggc
     1741 cctgcagctg gaggaggaga ggaagcgggc acaggaggag gccgagcgcc tagaggctga
     1801 ccgtatggct gcactgcggg ctaaggagga gctggagaga caggcggtgg atcagataaa
     1861 gagccaggag cagctggctg cggagcttgc agaatacact gccaagattg ccctcctgga
     1921 agaggcgcgg aggcgcaagg aggatgaagt tgaagagtgg cagcacaggg ccaaagaagc
     1981 ccaggatgac ctggtgaaga ccaaggagga gctgcacctg gtgatgacag cacccccgcc
     2041 cccaccaccc cccgtgtacg agccggtgag ctaccatgtc caggagagct tgcaggatga
     2101 gggcgcagag cccacgggct acagcgcgga gctgtctagt gagggcatcc gggatgaccg
     2161 caatgaggag aagcgcatca ctgaggcaga gaagaacgag cgtgtgcagc ggcagctgct
     2221 gacgctgagc agcgagctgt cccaggcccg agatgagaat aagaggaccc acaatgacat
     2281 catccacaac gagaacatga ggcaaggccg ggacaagtac aagacgctgc ggcagatccg
     2341 gcagggcaac accaagcagc gcatcgacga gttcgaggcc ctggcatccg gcggtagcgg
     2401 cggtggcagc ggaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
     2461 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     2521 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     2581 caagctgccc gtgccctggc ccaccctcgt gaccaccttg acctacggcg tgcagtgctt
     2641 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     2701 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     2761 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     2821 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaaggtcta
     2881 tatcaccgcc gacaagcaga agaacggcat caaggtgaac ttcaagaccc gccacaacat
     2941 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     3001 ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc
     3061 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     3121 cggcatggac gagctgtaca agtaagcggc cgcgactcta gatcataatc agccatacca
     3181 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3241 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3301 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3361 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     3421 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3481 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3541 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3601 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3661 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3721 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3781 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3841 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3901 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3961 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4021 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4081 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4141 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4201 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4261 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4321 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4381 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4441 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4501 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     4561 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4621 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4681 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4741 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4801 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4861 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     4921 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4981 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5041 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5101 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5161 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5221 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5281 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5341 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     5401 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     5461 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     5521 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     5581 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     5641 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5701 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5761 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5821 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5881 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5941 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6001 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6061 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6121 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6181 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6241 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6301 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6361 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6421 acatgttctt tcctgcgtta tcccctgatt ctgtgg
//
