LOCUS       sequence-483079-        5995 bp    DNA     circular SYN 13-AUG-2025
DEFINITION  Note- This plasmid contains Keratin 18. Localization: Cytokeratin, 
            Excitation: 488, Emission: 507.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5995)
  TITLE     Davidson Green fluorescent proteins
REFERENCE   2  (bases 1 to 5995)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 13, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: mEGFP-Keratin-17
FEATURES             Location/Qualifiers
     source          1..5995
                     /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      1875..1884
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1881..1883,1884..1886,1887..2600)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1926..1947)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2187..2206)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2534..2555
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2723..2844
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2760..2779)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2814..2833
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2851..3306)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2988..3009)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3200..3219
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3333..3437
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3439..3796
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3439..3630
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3462..3482)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3647..3782
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3831..4625
                     /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             3831..4625
                     /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(3885..3904)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4495..4514
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4813..4832)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4857..4904
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5233..5821
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5722..5741
                     /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 gctagcacca
      541 tgagcttcac cactcgctcc accttctcca ccaactaccg gtccctgggc tctgtccagg
      601 cgcccagcta cggcgcccgg ccggtcagca gcgcggccag cgtctatgca ggcgctgggg
      661 gctctggttc ccggatctcc gtgtcccgct ccaccagctt caggggcggc atggggtccg
      721 ggggcctggc caccgggata gccgggggtc tggcaggaat gggaggcatc cagaacgaga
      781 aggagaccat gcaaagcctg aacgaccgcc tggcctctta cctggacaga gtgaggagcc
      841 tggagaccga gaaccggagg ctggagagca aaatccggga gcacttggag aagaagggac
      901 cccaggtcag agactggagc cattacttca agatcatcga ggacctgagg gctcagatct
      961 tcgcaaatac tgtggacaat gcccgcatcg ttctgcagat tgacaatgcc cgtcttgctg
     1021 ctgatgactt tagagtcaag tatgagacag agctggccat gcgccagtct gtggagaacg
     1081 acatccatgg gctccgcaag gtcattgatg acaccaatat cacacgactg cagctggaga
     1141 cagagatcga ggctctcaag gaggagctgc tcttcatgaa gaagaaccac gaagaggaag
     1201 taaaaggcct acaagcccag attgccagct ctgggttgac cgtggaggta gatgccccca
     1261 aatctcagga cctcgccaag atcatggcag acatccgggc ccaatatgac gagctggctc
     1321 ggaagaaccg agaggagcta gacaagtact ggtctcagca gattgaggag agcaccacag
     1381 tgctcaccac acagtctgct gaggttggag ctgctgagac gacgctcaca gagctgagac
     1441 gtacagtcca gtccttggag atcgacctgg actccatgag aaatctgaag gccagcttgg
     1501 agaacagcct gagggaggtg gaggcccgct acgccctaca gatggagcag ctcaacggga
     1561 tcctgctgca ccttgagtca gagctggcac agacccgggc agagggacag cgccaggccc
     1621 aggagtatga ggccctgctg aacatcaagg tcaagctgga ggctgagatc gccacctacc
     1681 gccgcctgct ggaagatggc gaggacttta atcttggtga tgccttggac agcagcaact
     1741 ccatgcaaac catccaaaag accaccaccc gccggatagt ggatggcaaa gtggtgtctg
     1801 agaccaatga caccaaagtt ctgaggcatc gaattctgca gtcgacggta ccgcgggccc
     1861 gggatccacc ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     1921 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1981 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     2041 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     2101 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     2161 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     2221 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     2281 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     2341 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     2401 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2461 tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg
     2521 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2581 tggacgagct gtacaagtaa agcggccgcg actctagatc ataatcagcc ataccacatt
     2641 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2701 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     2761 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2821 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     2881 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     2941 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     3001 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3061 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     3121 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3181 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     3241 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     3301 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3361 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3421 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     3481 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3541 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3601 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3661 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3721 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3781 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3841 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3901 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3961 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     4021 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4081 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4141 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4201 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4261 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4321 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     4381 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4441 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4501 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4561 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4621 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4681 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4741 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     4801 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     4861 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     4921 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4981 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     5041 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     5101 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5161 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5221 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5281 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5341 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5401 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5461 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5521 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5581 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5641 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5701 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5761 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5821 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5881 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt
     5941 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccg
//
