LOCUS       sequence-402837-        5949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression in mammalian cells or oocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5949)
  AUTHORS   Abderemane-Ali F, Rossen ND, Kobiela ME, Craig RA, Garrison CE, Chen
            Z, Colleran CM, O'Connell LA, Du Bois J, Dumbacher JP, Minor DL
  TITLE     Evidence that toxin resistance in poison birds and frogs is not 
            rooted in sodium channel mutations and may rely on ""toxin sponge"" 
            proteins.
  JOURNAL   J Gen Physiol. 2021 Sep 6;153(9):e202112872. doi: 
            10.1085/jgp.202112872. Epub 2021 Aug 5.
  PUBMED    34351379
REFERENCE   2  (bases 1 to 5949)
  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: "J Gen 
            Physiol."; date: "2021-09-6"; volume: "153(9):e202112872. doi"; 
            pages: " 10.1085/jgp.202112872. Epub 2021 Aug 5"
COMMENT     Sequence Label: Pum NaVB2
FEATURES             Location/Qualifiers
     source          1..5949
                     /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"
     misc_feature    join(1248..1822,1823..1825,1826..1834)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1415..1432)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1642..1661
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1835..1837,1838..1840,1841..2554)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1835..1837,1838..1840,1841..2551)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1880..1901)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2141..2160)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2488..2509
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2677..2798
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2714..2733)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2768..2787
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2805..3260)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2942..2963)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3154..3173
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3287..3391
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3393..3750
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3393..3584
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3416..3436)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3601..3736
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3785..4579
                     /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             3785..4579
                     /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(3839..3858)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4449..4468
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4767..4786)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4811..4858
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5187..5775
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5676..5695
                     /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 atctcgagat gagccctgag gcatggctgc cacagcctac actgctgctg
      601 accggactga ccctgctgct gagtctggcc cctcctggac tgggaatgga agtgatggcc
      661 cctgccacaa tcaacgccct gaatggcagc tccgtgaagc tgtcctgcac cttcaactct
      721 tgttacaagg tggagaacaa gcagttttcc ctgaattgga catatcagga gtgctctaac
      781 tgtagcgagg agctgttcct gcagtttagg accaagatca tgaataagca gctggaccgg
      841 ttcggcaaca gagtggagtt taccggcaat cccgccaagt acgacgtgtc tttcacactg
      901 aagaatgtgc agctggagga tgagggcacc tacaactgct atgtgctgaa tccacctgac
      961 aggcacaggg gccacggcaa gatcagcctg aaggtgctga ccaaggagcc acccaagcac
     1021 gatagcacag tggccgtgat cgtgggagcc tccgtgggag gctttctggc cgtggtcatc
     1081 ctggtgctga tggtggtgaa gtgcgtgcgg agaaagaagc agcagagact gaacacagac
     1141 gatcagaaga ccgaagaaga aggcaaaacc gacggagagg gaaaccccga cgagggaaca
     1201 aaatgataag aattctgcag tcgacggtac cgcgggcccg ggatccgccc ctctccctcc
     1261 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     1321 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     1381 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     1441 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     1501 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     1561 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     1621 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     1681 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtacac atgctttaca
     1741 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     1801 ctttgaaaaa cacgatgata atatggccac aaccatggtg agcaagggcg aggagctgtt
     1861 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1921 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1981 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     2041 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     2101 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     2161 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     2221 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     2281 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     2341 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     2401 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     2461 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     2521 gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc
     2581 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2641 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2701 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2761 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     2821 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2881 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     2941 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     3001 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     3061 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     3121 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3181 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3241 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3301 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3361 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3421 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3481 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3541 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3601 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3661 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3721 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     3781 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3841 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3901 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3961 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4021 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4081 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4141 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4201 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4261 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     4321 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4381 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4441 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4501 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4561 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4621 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4681 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4741 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4801 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     4861 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     4921 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     4981 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     5041 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5101 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5161 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5221 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5281 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5341 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5401 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5461 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5521 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5581 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5641 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5701 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5761 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5821 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5881 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     5941 ggagttccg
//
