LOCUS       sequence-365707-        5845 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Focal Adhesions, Excitation: 487, Emission: 509.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5845)
  TITLE     Davidson mEmerald
REFERENCE   2  (bases 1 to 5845)
  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-Migfilin-N-16
FEATURES             Location/Qualifiers
     source          1..5845
                     /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      1726..1735
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1732..1734,1735..1737,1738..2451)
                     /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(1777..1798)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2038..2057)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2385..2406
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2573..2694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2610..2629)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2664..2683
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2701..3156)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2838..2859)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3050..3069
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3183..3287
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3289..3646
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3289..3480
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3312..3332)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3497..3632
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3681..4475
                     /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             3681..4475
                     /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(3735..3754)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4345..4364
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4663..4682)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4707..4754
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5083..5671
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5572..5591
                     /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 atctcgagac cgccatggcc tcaaagcctg agaagagggt ggcatcgtct
      601 gtctttatca ccctggcacc cccgcgccgc gatgtggccg tggcggagga agtgaggcag
      661 gcagtttgtg aggcccggcg tggccgcccc tgggaggctc ctgcccccat gaagacaccc
      721 gaggctggct tggcggggag gcccagcccc tggacaaccc ctggcagagc tgcagccaca
      781 gtgccggctg cacctatgca gctcttcaat ggaggatgcc caccccctcc tcctgtcctg
      841 gatggtgagg acgtgcttcc tgacctggac ctcctcccac cccctccacc gccccctcca
      901 gtgcttctgc cttctgaaga ggaggctcct gctccaatgg gggcctcact cattgcagac
      961 ttagagcagc tgcacctgtc cccgcccccg cccccaccac aggccccagc ggagggacct
     1021 tcagtccagc ccggtcccct caggcccatg gaggaagagc tgccacctcc cccggcagaa
     1081 cctgttgaga aaggggcatc cacagacatc tgtgccttct gctacaagac cgtgttcccc
     1141 cgagagctgg ctgtggaggc catgaagagg cagtaccatg cccagtgctt cacgtgccgc
     1201 acctgccgcc gccagctggc tgggcagagc ttctaccaga aggatgggcg acccctctgc
     1261 gaaccctgct accaggacac actggagagg tgcggcaagt gtggcgaggt ggtccgggac
     1321 cacatcatca gggccctggg ccaggccttc cacccctcct gcttcacgtg tgtgacctgc
     1381 gcccggtgca ttggggatga gagctttgcc ctgggcagcc agaacgaggt gtactgcctg
     1441 gacgacttct acaggaaatt cgcccccgtc tgcagcatct gtgaaaatcc catcatccct
     1501 cgggatggga aagatgcctt caaaatcgaa tgcatgggaa gaaacttcca tgaaaattgc
     1561 tacaggtgtg aggactgcag gatcctcctg tctgtcgagc ccacggacca aggctgctac
     1621 cccctgaaca accatctctt ctgcaagcca tgccatgtga agcggagtgc tgcggggtgc
     1681 tgcgcatccg ggggtgcgtc aggcggcagc ggatctggac cggtcgccac catggtgagc
     1741 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1801 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1861 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1921 accttgacct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     1981 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     2041 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     2101 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     2161 tacaactaca acagccacaa ggtctatatc accgccgaca agcagaagaa cggcatcaag
     2221 gtgaacttca agacccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     2281 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     2341 acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     2401 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta agcggccgcg
     2461 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2521 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2581 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2641 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     2701 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     2761 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     2821 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     2881 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     2941 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3001 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3061 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3121 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     3181 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3241 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     3301 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3361 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3421 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3481 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3541 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3601 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     3661 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3721 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3781 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3841 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     3901 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3961 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4021 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4081 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4141 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4201 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4261 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4321 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4381 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4441 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4501 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4561 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4621 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     4681 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     4741 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     4801 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     4861 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     4921 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4981 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5041 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5101 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5161 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5221 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5281 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5341 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5401 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5461 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5521 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5581 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5641 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5701 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5761 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
     5821 ttcatagccc atatatggag ttccg
//
