LOCUS       sequence-258728-        4903 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of GC localized oxBFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4903)
  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 4903)
  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: GalToxBFP
FEATURES             Location/Qualifiers
     source          1..4903
                     /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      784..793
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(790..792,793..795,796..1509)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFISTTGKLPVPWPTLVTTLSHGVQVFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(835..856)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    1631..1752
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1668..1687)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1722..1741
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1759..2214)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1896..1917)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2108..2127
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2241..2345
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2347..2704
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2347..2538
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2370..2390)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2555..2690
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2739..3533
                     /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             2739..3533
                     /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(2793..2812)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3403..3422
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3721..3740)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3765..3812
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4141..4729
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4630..4649
                     /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 atctcgagct cgccaccatg aggcttcggg agccgctcct gagcggcagc
      601 gccgcgatgc caggcgcgtc cctacagcgg gcctgccgcc tgctcgtggc cgtctgcgct
      661 ctgcaccttg gcgtcaccct cgtttactac ctggctggcc gcgacctgag ccgcctgccc
      721 caactggtcg gagtctccac accgctgcag tcgacggtac cgcgggcccg ggatccaccg
      781 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
      841 gagctggacg gcgacgtaaa cggccacaag ttcagcgtga ggggcgaggg cgagggcgat
      901 gccaccaacg gcaagctgac cctgaagttc atcagtacta ccggcaagct gcccgtgccc
      961 tggcccaccc tcgtgaccac cctgagccac ggcgtgcagg tgttcgcccg ctaccccgac
     1021 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1081 accatcttct tcaaggacga cggcacctac aagacccgcg ccgaggtgaa gttcgagggc
     1141 gacaccctgg tgaaccgcat cgagctgaag ggcgtcgact tcaaggagga cggcaacatc
     1201 ctggggcaca agctggagta caacttcaac agccacaaca tctatatcat ggccgtcaag
     1261 cagaagaacg gcatcaaggc caacttcaag atccgccaca acgtggagga cggcagcgtg
     1321 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1381 gacagccact acctgagcac ccagtccgtg ctgagcaaag accccaacga gaagcgcgat
     1441 cacatggtcc tgctggagtt ccgcaccgcc gccgggatca ctctcggcat ggacgagctg
     1501 tacaagtaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     1561 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     1621 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     1681 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     1741 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     1801 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     1861 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     1921 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     1981 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     2041 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     2101 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     2161 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     2221 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2281 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2341 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     2401 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     2461 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2521 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     2581 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     2641 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     2701 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     2761 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     2821 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     2881 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     2941 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3001 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3061 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3121 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3181 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3241 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     3301 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3361 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3421 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3481 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     3541 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     3601 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     3661 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     3721 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     3781 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     3841 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     3901 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     3961 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     4021 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4081 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4141 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4201 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4261 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4321 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4381 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4441 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4501 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4561 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4621 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4681 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4741 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     4801 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
     4861 caattacggg gtcattagtt catagcccat atatggagtt ccg
//
