LOCUS       sequence-230170-        6979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Microtubule, N1 Cloning Vector, Excitation:487, 
            Emission:509.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6979)
  TITLE     Michael Davidson Unpublished
REFERENCE   2  (bases 1 to 6979)
  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-Enconsin-N-18
FEATURES             Location/Qualifiers
     source          1..6979
                     /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      559..568
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2860..2869
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2866..2868,2869..2871,2872..3585)
                     /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(2911..2932)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3172..3191)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3519..3540
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3707..3828
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3744..3763)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3798..3817
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3835..4290)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3972..3993)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4184..4203
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4317..4421
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4423..4780
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4423..4614
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4446..4466)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4631..4766
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4815..5609
                     /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             4815..5609
                     /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(4869..4888)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5479..5498
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5797..5816)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5841..5888
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6217..6805
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6706..6725
                     /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 caccatggcg gagctaggag ctggcggcga cggccacagg
      601 ggcggcgacg gcgcagtgcg aagcgaaaca gcacccgaca gctacaaagt gcaagataag
      661 aaaaatgcct ccagccgccc tgcctctgca atttcaggac aaaataacaa ccactcagga
      721 aataaaccag accctccgcc tgtgttacgt gttgatgacc ggcagcggct ggcccgggag
      781 cgacgtgagg aacgggagaa acagctagct gcaagagaaa tagtgtggtt agaaagagaa
      841 gagcgagcca ggcagcacta cgagaagcac ctggaagagc ggaagaagag gttggaggag
      901 cagaggcaga aggaggagcg gaggagggct gctgtggagg agaagcggag gcagagactt
      961 gaggaggaca aagaacgcca cgaagctgtt gtacggcgca caatggaaag gagccagaag
     1021 ccaaaacaga agcataaccg ttggtcgtgg ggaggctctc tccatgggag ccctagcatc
     1081 cacagtgcag atccagacag gcggtcagtt tccaccatga atctttcgaa atatgttgat
     1141 cccgtcatta gcaagcggct ctcctcttca tctgcaactt tactaaattc tccagataga
     1201 gctcgccgcc tgcagctcag cccatgggag agcagcgttg ttaacagact cctgacgccc
     1261 acacattcgt tcctggccag aagtaaaagc acagctgcct tgtctggaga agcagcatct
     1321 tgcagcccca tcatcatgcc ctacaaagct gcacactcta gaaattcgat ggatcgacca
     1381 aaactctttg taacaccacc tgagggctct tctcgcagga ggatcattca tggcacagcg
     1441 agctataaaa aagaaagaga gagagaaaat gtactcttcc tcacatctgg cacccgaagg
     1501 gctgtatctc catctaatcc caaagcaaga caaccagctc gctcccgact ttggcttccg
     1561 tccaagtctc ttcctcattt gcctggcaca cccagaccga catcctcctt gccacccggc
     1621 tcagtcaaag ctgctcctgc tcaggtccgg cccccatccc ccggcaacat ccgccctgtc
     1681 aagagggaag tcaaagtgga gcctgagaag aaagatcctg agaaggaacc tcagaaagtt
     1741 gccaatgagc cctcactaaa gggcagagca cctttagtga aggtagaaga agccacagtt
     1801 gaagagcgga cacctgctga accagaagtt ggccctgctg ctccagccat ggccccagct
     1861 ccagcctcgg ccccagctcc agcctcggcc ccagctccag ccccggtccc caccccagcc
     1921 atggtctcag ccccgtcatc cactgtgaat gccagtgctt ctgttaagac ttctgcaggc
     1981 accaccgacc cagaggaggc cacaaggctt ctagctgaga agaggcggct ggcccgagag
     2041 cagagagaaa aggaagaaag ggagaggagg gagcaggaag agcttgaaag acaaaagaga
     2101 gaggaattgg ctcaacgtgt ggctgaagag aggacgactc gccgtgagga ggagtcgcgc
     2161 aggctggaag ccgagcaggc ccgggagaag gaggagcagc tgcagcggca ggcggaggag
     2221 cgggcgctgc gcgagtggga ggaggcagag cgcgcccaga ggcagaaaga agaagaagct
     2281 cgcgttcgtg aagaagcaga gagggtccgg caggaacgag agaagcattt ccagagagaa
     2341 gagcaagagc gcctggagag aaagaagcga cttgaggaga ttatgaaaag aaccaggaga
     2401 acagaagcta cagataagaa aaccagtgat cagagaaacg gtgatatagc caagggagct
     2461 ctcactggag gaacagaggt gtctgcactt ccatgtacaa caaacgctcc gggaaatgga
     2521 aagccagttg gtagcccaca tgtggttacc tcacaccagt caaaagtgac agtggagagc
     2581 actcccgatt tggaaaaaca accaaatgaa aatggtgtat ctgttcagaa tgaaaatttt
     2641 gaagaaatta taaacttacc cattggatct aaaccatcca gattagatgt caccaacagt
     2701 gagagcccag aaattccttt gaatccaatt ttggcctttg atgatgaagg gacacttggg
     2761 cccctgcctc aggtagatgg tgttcagaca cagcagactg cagaagttat aggcagcgca
     2821 ggcggaagcg gggtaccgcg ggcccgggat ccaccggtcg ccaccatggt gagcaagggc
     2881 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2941 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     3001 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttg
     3061 acctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     3121 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     3181 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     3241 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     3301 tacaacagcc acaaggtcta tatcaccgcc gacaagcaga agaacggcat caaggtgaac
     3361 ttcaagaccc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     3421 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     3481 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     3541 accgccgccg ggatcactct cggcatggac gagctgtaca agtaagcggc cgcgactcta
     3601 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     3661 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3721 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3781 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg
     3841 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3901 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     3961 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     4021 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     4081 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     4141 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     4201 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     4261 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     4321 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4381 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     4441 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4501 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4561 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4621 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4681 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     4741 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     4801 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4861 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4921 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4981 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5041 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5101 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5161 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5221 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5281 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5341 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5401 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5461 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5521 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5581 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5641 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5701 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5761 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     5821 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     5881 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     5941 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     6001 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     6061 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6121 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6181 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6241 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6301 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6361 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6421 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6481 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6541 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6601 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6661 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6721 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6781 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6841 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6901 ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata
     6961 gcccatatat ggagttccg
//
