LOCUS       sequence-393245-        6462 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Nuclear Envelope, Excitation: 383, Emission: 448.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6462)
  TITLE     Davidson Blue Fluorescent Proteins
REFERENCE   2  (bases 1 to 6462)
  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-LaminB1-10
FEATURES             Location/Qualifiers
     source          1..6462
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        42..399
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        42..233
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(65..85)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      250..385
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             434..1228
                     /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             434..1228
                     /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(488..507)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1098..1117
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1416..1435)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1460..1507
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1836..2424
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2325..2344
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2599..2902
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2903..3106
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3057..3077
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3145..3154
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3151..3153,3154..3156,3157..3867)
                     /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(3196..3217)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    5788..5909
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5825..5844)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5879..5898
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5916..6371)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6053..6074)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6265..6284
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(6398..6462,1..40)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
       61 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
      121 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
      181 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
      241 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
      301 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
      361 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
      421 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
      481 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
      541 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
      601 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
      661 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
      721 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
      781 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
      841 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
      901 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
      961 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     1021 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     1081 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     1141 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     1201 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     1261 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     1321 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     1381 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     1441 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     1501 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     1561 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     1621 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     1681 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1741 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     1801 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1861 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1921 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1981 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2041 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2101 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2161 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2221 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2281 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2341 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2401 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2461 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     2521 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
     2581 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     2641 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     2701 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     2761 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     2821 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     2881 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     2941 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     3001 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     3061 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
     3121 tcagatccgc tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
     3181 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     3241 aggggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt catctgcacc
     3301 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgagcca cggcgtgcag
     3361 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     3421 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaccta caagacccgc
     3481 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcgtcgac
     3541 ttcaaggagg acggcaacat cctggggcac aagctggagt acaacttcaa cagccacaac
     3601 atctatatca tggccgtcaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     3661 aacgtggagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     3721 gacggccccg tgctgctgcc cgacagccac tacctgagca cccagtccgt gctgagcaaa
     3781 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tccgcaccgc cgccgggatc
     3841 actctcggca tggacgagct gtacaagtcc ggactcagat ctcgagctca agcttcgatg
     3901 gcgactgcga cccccgtgcc gccgcggatg ggcagccgcg ctggcggccc caccacgccg
     3961 ctgagcccca cgcgcctgtc gcggctccag gagaaggagg agctgcgcga gctcaatgac
     4021 cggctggcgg tgtacatcga caaggtgcgc agcctggaga cggagaacag cgcgctgcag
     4081 ctgcaggtga cggagcgcga ggaggtgcgc ggccgtgagc tcaccggcct caaggcgctc
     4141 tacgagaccg agctggccga cgcgcgacgc gcgctcgacg acacggcccg cgagcgcgcc
     4201 aagctgcaga tcgagctggg caagtgcaag gcggaacacg accagctgct cctcaactat
     4261 gctaagaagg aatctgatct taatggcgcc cagatcaagc ttcgagaata tgaagcagca
     4321 ctcaattcga aagatgcagc tcttgctact gcacttggtg acaaaaaaag tttagaggga
     4381 gatttggagg atctgaagga tcagattgcc cagttggaag cctccttagc tgcagccaaa
     4441 aaacagttag cagatgaaac tttacttaaa gtagatttgg agaatcgttg tcagagcctt
     4501 actgaggact tggagtttcg caaaagcatg tatgaagagg agattaacga gaccagaagg
     4561 aagcatgaaa cgcgcttggt agaggtggat tctgggcgtc aaattgagta tgagtacaag
     4621 ctggcgcaag cccttcatga gatgagagag caacatgatg cccaagtgag gctgtataag
     4681 gaggagctgg agcagactta ccatgccaaa cttgagaatg ccagactgtc atcagagatg
     4741 aatacttcta ctgtcaacag tgccagggaa gaactgatgg aaagccgcat gagaattgag
     4801 agcctttcat cccagctttc taatctacag aaagagtcta gagcatgttt ggaaaggatt
     4861 caagaattag aggacttgct tgctaaagaa aaagacaact ctcgtcgcat gctgacagac
     4921 aaagagagag agatggcgga aataagggat caaatgcagc aacagctgaa tgactatgaa
     4981 cagcttcttg atgtaaagtt agccctggac atggaaatca gtgcttacag gaaactctta
     5041 gaaggcgaag aagagaggtt gaagctgtct ccaagccctt cttcccgtgt gacagtatcc
     5101 cgagcatcct caagtcgtag tgtacgtaca actagaggaa agcggaagag ggttgatgtg
     5161 gaagaatcag aggcgagtag tagtgttagc atctctcatt ccgcctcagc cactggaaat
     5221 gtttgcatcg aagaaattga tgttgatggg aaatttatcc gcttgaagaa cacttctgaa
     5281 caggatcaac caatgggagg ctgggagatg atcagaaaaa ttggagacac atcagtcagt
     5341 tataaatata cctcaagata tgtgctgaag gcaggccaga ctgttacaat ttgggctgca
     5401 aacgctggtg tcacagccag ccccccaact gacctcatct ggaagaacca gaactcgtgg
     5461 ggcactggcg aagatgtgaa ggttatattg aaaaattctc agggagagga ggttgctcaa
     5521 agaagtacag tctttaaaac aaccatacct gaagaagagg aggaggagga agaagcagct
     5581 ggagtggttg ttgaggaaga acttttccac cagcagggaa ccccaagagc atccaataga
     5641 agctgtgcaa ttatgtaggg atccaccgga tctagataac tgatcataat cagccatacc
     5701 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     5761 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     5821 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5881 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
     5941 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     6001 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     6061 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     6121 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     6181 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     6241 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     6301 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     6361 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6421 tttctaaata cattcaaata tgtatccgct catgagacaa ta
//
