LOCUS       sequence-305814-        6938 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Membrane Scaffold, Excitation: 487, Emission: 509.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6938)
  TITLE     Davidson mEmerald
REFERENCE   2  (bases 1 to 6938)
  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-PSD95-C-16
FEATURES             Location/Qualifiers
     source          1..6938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        99..402
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        403..606
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     557..577
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      645..654
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(651..653,654..656,657..1367)
                     /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(696..717)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(957..976)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1304..1325
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3588..3640
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3764..3885
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3801..3820)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3855..3874
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3892..4347)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4029..4050)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4241..4260
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4374..4478
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4480..4837
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4480..4671
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4503..4523)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4688..4823
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4872..5666
                     /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             4872..5666
                     /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(4926..4945)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5536..5555
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5854..5873)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5898..5945
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6274..6862
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6763..6782
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt
       61 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      121 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      181 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      241 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      301 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      361 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      421 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      481 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      541 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      601 agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggtcgccacc atggtgagca
      661 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      721 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      781 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      841 ccttgaccta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
      901 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
      961 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1021 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1081 acaactacaa cagccacaag gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     1141 tgaacttcaa gacccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     1201 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1261 cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1321 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc ggagggagcg
     1381 gcagtgggtc cagtggaggg agcagtggaa gcggagactg tctctgtata gtgacaacca
     1441 agaaataccg ctaccaagat gaagacacgc cccctctgga acacagcccg gcccacctcc
     1501 ccaaccaggc caattctccc cctgtgattg tcaacacgga caccctagaa gccccaggat
     1561 atgagttgca ggtgaatgga acagaggggg agatggagta tgaggagatc acattggaaa
     1621 ggggtaactc aggtctgggc ttcagcatcg caggtggcac tgacaacccg cacatcggtg
     1681 acgacccgtc catttttatc accaagatca ttcctggtgg ggctgcagcc caggatggcc
     1741 gcctcagggt caatgacagc atcctgtttg taaatgaagt ggatgttcgg gaggtgaccc
     1801 attcagctgc ggtggaggcc ctcaaagagg caggttccat cgttcgcctc tatgtcatgc
     1861 gccggaaacc cccagccgaa aaggtcatgg agatcaaact catcaaaggg cctaaaggac
     1921 ttggcttcag cattgcgggg ggcgttggga accagcacat ccctggagat aacagcatct
     1981 atgtaacgaa gatcatcgaa ggaggtgctg cccacaagga tggcaggttg cagattggag
     2041 acaagatcct ggcggtcaac agtgtggggc tggaggacgt catgcacgag gatgccgtgg
     2101 cagccctgaa gaacacatat gacgttgtgt acctaaaggt ggccaagccc agcaatgcct
     2161 acctgagtga cagctatgct cccccagaca tcacaacctc gtattctcag cacctggaca
     2221 atgagatcag tcatagcagc tacttgggca ctgactaccc cacagccatg acccccactt
     2281 cccctcggcg ctactcccct gtggccaagg acctgctggg ggaggaagac attccccggg
     2341 aaccaaggcg gatcgtgatc catcggggct ccaccggcct gggcttcaac atcgtgggcg
     2401 gcgaggatgg tgaaggcatc ttcatctcct tcatccttgc tgggggtcca gccgacctca
     2461 gtggggagct acggaagggg gaccagatcc tgtcggtcaa tggtgttgac ctccgcaatg
     2521 ccagtcacga acaggctgcc attgccctga agaatgcggg tcagacggtc acgatcatcg
     2581 ctcagtataa accagaagag tatagtcgat tcgaggccaa gatccatgat cttcgggaac
     2641 agctcatgaa tagtagccta ggctcaggga ctgcatcctt gcgaagcaac cccaagaggg
     2701 gcttctacat tagggccctg tttgattacg acaagaccaa ggactgcggt ttcttgagcc
     2761 aggccctgag cttccgcttc ggggatgtgc ttcatgtcat tgacgctggt gacgaagagt
     2821 ggtggcaagc acggcgggtc cactccgaca gtgagaccga cgacattggc ttcattccca
     2881 gcaaacggcg ggtcgagcga cgagagtggt caaggttaaa ggccaaggac tggggctcca
     2941 gctctggatc acagggtcga gaagactcgg ttctgagcta tgagacggtg acccagatgg
     3001 aagtgcacta tgctcgtccc atcatcatcc ttggacccac caaagaccgt gccaacgatg
     3061 atcttctctc cgagttcccc gacaagtttg gatcctgtgt ccctcatacg acacgtccta
     3121 agcgggaata tgagatagac ggccgggatt accactttgt ctcctcccgg gagaaaatgg
     3181 agaaggacat ccaggcacac aagttcattg aggctggcca gtacaacagc cacctctatg
     3241 ggaccagcgt ccagtctgtg cgagaggtag cagagcaggg gaagcactgc atcctcgatg
     3301 tctcggccaa tgccgtgcgg cggctgcagg cggcccacct gcaccccatc gccatcttca
     3361 tccgtccccg ctccctggag aatgtgctag agatcaataa gcggatcaca gaggagcaag
     3421 cccggaaagc cttcgacaga gccacgaagc tggagcagga gttcacagag tgcttctcag
     3481 ccatcgtaga gggcgacagc tttgaagaga tctatcacaa agtgaaacgt gtcattgaag
     3541 acctctcagg cccctacatc tgggtcccag cccgagagag actctgactc gagctcaagc
     3601 ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc accggatcta gataactgat
     3661 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     3721 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3781 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3841 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa
     3901 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3961 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     4021 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     4081 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     4141 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     4201 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     4261 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     4321 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4381 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4441 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     4501 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4561 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4621 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4681 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4741 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4801 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     4861 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4921 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4981 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     5041 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     5101 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     5161 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5221 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5281 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5341 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     5401 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5461 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5521 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5581 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5641 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5701 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5761 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5821 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     5881 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     5941 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     6001 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     6061 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     6121 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6181 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6241 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6301 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6361 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6421 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6481 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6541 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6601 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6661 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6721 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6781 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6841 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6901 ttgctggcct tttgctcaca tgttctttcc tgcgttat
//
