LOCUS       sequence-364377-        4732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-terminal fusion to oxBFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4732)
  AUTHORS   Costantini LM, Baloban M, Markwardt ML, Rizzo M, Guo F, Verkhusha 
            VV, Snapp EL
  TITLE     A palette of fluorescent proteins optimized for diverse cellular 
            environments.
  JOURNAL   Nat Commun. 2015 Jul 9;6:7670. doi: 10.1038/ncomms8670.
  PUBMED    26158227
REFERENCE   2  (bases 1 to 4732)
  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"; journalName: "Nat 
            Commun."; date: "2015-07-9"; volume: "6:7670. doi"; pages: " 
            10.1038/ncomms8670"
COMMENT     Sequence Label: moxBFP
FEATURES             Location/Qualifiers
     source          1..4732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        12..203
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     160..180
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(205..309)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      336..791
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(423..442)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     633..654
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    798..919
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(809..828)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     863..882
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1041..1754,1755..1757,1758..1760))
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFISTTGKLPVPWPTLVTTLSHGVQVFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSKLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     1694..1715
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1757..1766
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1768..1848
                     /label=MCS
                     /note="multiple cloning site"
     promoter        complement(1871..2074)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1900..1920)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2075..2378
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2553..3141)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2633..2652)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3470..3517
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3542..3561
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3749..4543)
                     /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             complement(3749..4543)
                     /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(3860..3879)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4470..4489
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(4578..4732,1..203))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4592..4727
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc
       61 ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt
      121 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc
      181 acagctggtt ctttccgcct caggactctt cctttttcaa tattattgaa gcatttatca
      241 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
      301 ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag
      361 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
      421 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
      481 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
      541 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
      601 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
      661 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
      721 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
      781 gcttacaatt tacgccttaa gatacattga tgagtttgga caaaccacaa ctagaatgca
      841 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
      901 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
      961 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta
     1021 tgatctagag tcgcggccgc ttacttgtac agctcgtcca tgccgagagt gatcccggcg
     1081 gcggtgcgga actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctaagc
     1141 ttggactggg tgctcaggta gtggctgtcg ggcagcagca cggggccgtc gccgatgggg
     1201 gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctccacgtt gtggcggatc
     1261 ttgaagttgg ccttgatgcc gttcttctgc ttgacggcca tgatatagat gttgtggctg
     1321 ttgaagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgacgccc
     1381 ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg
     1441 taggtgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg
     1501 gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc gggcgaacac ctgcacgccg
     1561 tggctcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt agtactgatg
     1621 aacttcaggg tcagcttgcc gttggtggca tcgccctcgc cctcgcccct cacgctgaac
     1681 ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc
     1741 tcctcgccct tgctcaccat ggtggcgacc ggtggatccc gggcccgcgg taccgtcgac
     1801 tgcagaattc gaagcttgag ctcgagatct gagtccggta gcgctagcgg atctgacggt
     1861 tcactaaacc agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg
     1921 tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca
     1981 aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc
     2041 cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt
     2101 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg
     2161 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta
     2221 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
     2281 ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg
     2341 gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat
     2401 gaactaatga ccccgtaatt gattactatt aataactaat gcatggcggt aatacggtta
     2461 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2521 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2581 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2641 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2701 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2761 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2821 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2881 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2941 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3001 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3061 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3121 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3181 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3241 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaacct
     3301 gaggctatgg cagggcctgc cgccccgacg ttggctgcga gccctgggcc ttcacccgaa
     3361 cttggggggt ggggtgggga aaaggaagaa acgcgggcgt attggcccca atggggtctc
     3421 ggtggggtat cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac
     3481 ccaacaccgt gcgttttatt ctgtcttttt attgccgtca tagcgcgggt tccttccggt
     3541 attgtctcct tccgtgtttc agttagcctc cccctagggt gggcgaagaa ctccagcatg
     3601 agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac
     3661 ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg
     3721 tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg
     3781 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
     3841 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg
     3901 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
     3961 tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct
     4021 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
     4081 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt
     4141 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
     4201 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc
     4261 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa
     4321 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac
     4381 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg
     4441 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
     4501 aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc
     4561 ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
     4621 tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
     4681 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cg
//
