LOCUS       sequence-161453-        6801 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Danio rerio VSP with separated EGFP in mammalian cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6801)
  AUTHORS   Hossain MI, Iwasaki H, Okochi Y, Chahine M, Higashijima S, Nagayama 
            K, Okamura Y
  TITLE     Enzyme domain affects the movement of the voltage sensor in ascidian
            and zebrafish voltage-sensing phosphatases.
  JOURNAL   J Biol Chem. 2008 Jun 27;283(26):18248-59. doi: 
            10.1074/jbc.M706184200. Epub 2008 Mar 28.
  PUBMED    18375390
REFERENCE   2  (bases 1 to 6801)
  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"; doi: 
            "10.1074/jbc.M706184200"; journalName: "J Biol Chem"; date: 
            "2008-06-27- 27"; volume: "283"; issue: "26"; pages: "18248-59"
COMMENT     Sequence Label: pDrVSP-IRES2-EGFP
FEATURES             Location/Qualifiers
     source          1..6801
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        113..416
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        417..620
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     571..591
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(2212..2786,2787..2789,2790..2798)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2379..2396)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2606..2625
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2799..2801,2802..2804,2805..3518)
                     /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(2799..2801,2802..2804,2805..3515)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2844..2865)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3105..3124)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3452..3473
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3641..3762
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3678..3697)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3732..3751
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3769..4224)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3906..3927)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4118..4137
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4251..4355
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4357..4714
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4357..4548
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4380..4400)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4565..4700
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4749..5543
                     /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             4749..5543
                     /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(4803..4822)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5413..5432
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5731..5750)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5775..5822
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6151..6739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6640..6659
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
       61 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      121 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      181 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      241 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      301 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      361 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      421 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      481 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      541 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      601 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagagc cgccaccatg
      661 acgtctgtgc attttaaccc tgggttagat tccaaagaag ttaatggtaa ttcggtgaaa
      721 gaagaggcag aggtgcagat cggtgatgga aaggaagaaa ccaaagaccc agacacaatg
      781 taccatcagg tccggaaaaa gataactcct tttgtgatgt cctttggttt tcgtgtattt
      841 ggcctggtac tgatcatctt ggacatcatt atggtcattg tggacttgtc tctttctgag
      901 aagagtcgtg atgttggagg tgcactggag actgtttctc tggtcatctc tttctttttc
      961 ctcatcgatg tattgctccg tgtttatgtg gagggattca aagtgtattt cagctcaaaa
     1021 ctgaatatag tagatgcttg tattgtggtc attactctgg tggtcaccat gatctacgca
     1081 ttctctgatt tctcaggagc cagtctgatt cccagggtgg tgacattcct gaggtctctg
     1141 aggatcctaa ttctggtacg catcttcaga ctggcctcac agaagagaga gcttgaaaaa
     1201 gtcactagga gaatggtttc agagaacaaa agacggtacc aaaaagatgg atttgatctg
     1261 gacctcacat acgtcacaga aagagtcatc gccatgtctt ttccatcttc tgggaagcag
     1321 gcgctttata ggaacccaat cagagaggtc gttagattct tggacaccaa acatatggat
     1381 cattacaagg tttttaatct ctgcagtgag aaaggttatg atccaaagtt ctttcactac
     1441 agagtagagc gtgtcatgat agatgaccat aatgtccctt cactggatga tatgctgcgc
     1501 tacaccgcct gtgtcagaga ctggatggca gcagactcca ggaatgttat tgccatccac
     1561 tgcaaaggag gaaagggtcg gaccggcact atggtttgca catggctcat tgacagcgat
     1621 cagtttgaga gtgcacagga gagcctggat tactttggag agagacggac tgataaaagc
     1681 atgagctcca agtttcaagg tgttgaaact ccgtctcaga gcaggtatgt tggctattat
     1741 gagatcatga agaatcagta caaccgtcag cttcctcctc ggaagagcct gaaaatcaag
     1801 agcatccgaa tccactccat cgcaggtgtg gggaagggca acggcagtga cctgaaactg
     1861 aagatcatag tgaagcacga gcttgttttc cagtgtgttt gtgccaaaca gcacaactgt
     1921 acagtgtttc ctgacactgg tagcaatgcg gtggtcatca gtttacagga cgggcctata
     1981 gtgactggag atgtgaaggt catgtttgag tccagtgctg gtttgccgaa gggttatgag
     2041 gactgtccct tctatttctg gttcaacacc tcatttgtgg agaactacag actttttctg
     2101 tcgagggaag aactggacaa tcctcacaaa cccaagacct gggacattta caaggaagac
     2161 tttggagtga cgctgtcttt cactgagcct tgaccgcggg cccgggatcc gcccctctcc
     2221 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
     2281 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     2341 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     2401 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
     2461 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     2521 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     2581 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     2641 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt acacatgctt
     2701 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
     2761 tttcctttga aaaacacgat gataatatgg ccacaaccat ggtgagcaag ggcgaggagc
     2821 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     2881 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     2941 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     3001 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     3061 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     3121 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     3181 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     3241 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     3301 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     3361 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     3421 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     3481 ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat
     3541 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3601 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3661 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3721 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     3781 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3841 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3901 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3961 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     4021 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     4081 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     4141 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     4201 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4261 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4321 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     4381 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4441 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4501 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4561 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4621 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4681 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     4741 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4801 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4861 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4921 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4981 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     5041 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     5101 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     5161 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     5221 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     5281 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5341 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5401 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5461 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5521 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5581 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5641 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5701 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5761 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5821 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5881 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     5941 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     6001 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6061 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6121 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6181 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6241 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6301 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6361 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6421 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6481 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6541 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6601 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6661 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6721 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6781 ctggcctttt gctcacatgt t
//
