LOCUS       sequence-230171-        5589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Nucleus, Excitation: 383, Emission: 448.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5589)
  TITLE     Davidson Blue Fluorescent Proteins
REFERENCE   2  (bases 1 to 5589)
  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: EBFP2-PCNA-19-SV40NLS-4
FEATURES             Location/Qualifiers
     source          1..5589
                     /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      535..544
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             547..567
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             571..591
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             595..615
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      622..631
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(628..630,631..633,634..1344)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(673..694)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2317..2438
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2354..2373)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2408..2427
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2445..2900)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2582..2603)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2794..2813
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2927..3031
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3033..3390
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3033..3224
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3056..3076)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3241..3376
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3425..4219
                     /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             3425..4219
                     /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(3479..3498)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4089..4108
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4407..4426)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4451..4498
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4827..5415
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5316..5335
                     /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 gctagccacc
      541 atggatccaa aaaagaagag aaaggtagat ccaaaaaaga agagaaaggt agatccaaaa
      601 aagaagagaa aggtaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
      661 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgagg
      721 ggcgagggcg agggcgatgc caccaacggc aagctgaccc tgaagttcat ctgcaccacc
      781 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgagccacgg cgtgcagtgc
      841 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
      901 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcacctacaa gacccgcgcc
      961 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg cgtcgacttc
     1021 aaggaggacg gcaacatcct ggggcacaag ctggagtaca acttcaacag ccacaacatc
     1081 tatatcatgg ccgtcaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     1141 gtggaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1201 ggccccgtgc tgctgcccga cagccactac ctgagcaccc agtccgtgct gagcaaagac
     1261 cccaacgaga agcgcgatca catggtcctg ctggagttcc gcaccgccgc cgggatcact
     1321 ctcggcatgg acgagctgta caagtccgga gaagggcaag ggcaagggca agggccgggc
     1381 cgcggctacg cgtatcgatc catgttcgag gcgcgcctgg tccagggctc catcctcaag
     1441 aaggtgttgg aggcactcaa ggacctcatc aacgaggcct gctgggatat tagctccagc
     1501 ggtgtaaacc tgcagagcat ggactcgtcc cacgtctctt tggtgcagct caccctgcgg
     1561 tctgagggct tcgacaccta ccgctgcgac cgcaacctgg ccatgggcgt gaacctcacc
     1621 agtatgtcca aaatactaaa atgcgccggc aatgaagata tcattacact aagggccgaa
     1681 gataacgcgg ataccttggc gctagtattt gaagcaccaa accaggagaa agtttcagac
     1741 tatgaaatga agttgatgga tttagatgtt gaacaacttg gaattccaga acaggagtac
     1801 agctgtgtag taaagatgcc ttctggtgaa tttgcacgta tatgccgaga tctcagccat
     1861 attggagatg ctgttgtaat ttcctgtgca aaagacggag tgaaattttc tgcaagtgga
     1921 gaacttggaa atggaaacat taaattgtca cagacaagta atgtcgataa agaggaggaa
     1981 gctgttacca tagagatgaa tgaaccagtt caactaactt ttgcactgag gtacctgaac
     2041 ttctttacaa aagccactcc actctcttca acggtgacac tcagtatgtc tgcagatgta
     2101 ccccttgttg tagagtataa aattgcggat atgggacact taaaatacta cttggctccc
     2161 aagatcgagg atgaagaagg atcttaggga tccaccggat ctagataact gatcataatc
     2221 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2281 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2341 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2401 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
     2461 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2521 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     2581 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     2641 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     2701 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     2761 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     2821 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     2881 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2941 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3001 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3061 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3121 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3181 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3241 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3301 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3361 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     3421 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3481 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3541 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3601 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3661 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3721 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3781 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3841 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3901 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     3961 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4021 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4081 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4141 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4201 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4261 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4321 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4381 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4441 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     4501 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     4561 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     4621 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     4681 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4741 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4801 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4861 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4921 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4981 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5041 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5101 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5161 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5221 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5281 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5341 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5401 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5461 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5521 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     5581 ggagttccg
//
