LOCUS       sequence-463617-        7455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mouse Kcnb2 fused with EGFP in C-terminal..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7455)
  TITLE     Sesti lab plasmids 2024
REFERENCE   2  (bases 1 to 7455)
  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: GFP-Kv2.2
FEATURES             Location/Qualifiers
     source          1..7455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(151..170)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     498..519
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3475..3596
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3512..3531)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3566..3585
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3603..4058)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3740..3761)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3952..3971
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4085..4189
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4191..4548
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4191..4382
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4214..4234)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4399..4534
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4583..5377
                     /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             4583..5377
                     /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(4637..4656)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5247..5266
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5565..5584)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5609..5656
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5985..6573
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6474..6493
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6748..7051
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7052..7255
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7206..7226
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      7294..7303
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7300..7302,7303..7305,7306..7455,1..561)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7300..7302,7303..7305,7306..7455,1..561)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7345..7366)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
       61 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      121 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
      181 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
      241 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
      301 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
      361 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
      421 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
      481 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
      541 ggcatggacg agctgtacaa gtccggactc agatctcgag ctcaagcttc gaattctgca
      601 gtcgacatgg cagaaaaggc acctcctggc ttgaacagaa aaacatccag gtcaacactt
      661 tctcttcctc cagaaccagt ggacattatc cgaagcaaaa catgctctcg gagagttaag
      721 atcaacgtag ggggccttaa ccacgaagtc ctgtggagaa ctctggacag gctacccagg
      781 acgcgcctgg ggaagctccg agactgtaac acgcacgaga gtctcctgga ggtgtgcgat
      841 gactacaacc ttaatgagaa tgagtatttc ttcgaccgcc atcctggagc cttcacatct
      901 attctgaact tctaccggac gggaaagctc cacatgatgg aagaaatgtg tgccctctcc
      961 tttggccaag agcttgatta ctggggcatc gatgagatct acctagagtc ctgctgccaa
     1021 gccagatacc atcagaagaa ggagcagatg aacgaagaac ttagaaggga ggcagagacc
     1081 atgcgtgagc gcgaggggga ggagtttgat aacacctgct gcccagagaa aaggaagaaa
     1141 ctttgggact tgctggagaa gccgaactca tccgtggcgg caaagatcct ggccatcgtg
     1201 tctatcctgt tcatcgtcct ttccaccatt gctctgtctc tcaacacact tccggagctg
     1261 caagaaaacg atgaatttgg gcaacccagt gacaaccgga agttggcgca cgtagaggct
     1321 gtgtgcattg cttggtttac catggagtac cttttacgat tcctgtcctc accaaataaa
     1381 tggaaattct ttaaaggccc actgaatgtc attgacctcc tggccatctt gccatactat
     1441 gtaaccatct tcctcaccga gtccaacaaa agtgtgctgc agttccagaa tgtgagacgc
     1501 gtggtccaga tctttcgaat catgcgcatc ctcaggatcc tgaaactcac cagacactcg
     1561 acgggcctgc agtctctggg tttcaccctc aggcggagtt acaatgagtt gggtttatta
     1621 atactgtttc tggccatggg aataatgata ttttccagtt tggtgttttt cgctgagaaa
     1681 gatgaagatg ctacaaagtt caccagtatc cctgcatcat tttggtgggc taccatcacc
     1741 atgaccactg tgggctatgg tgacatttac cctaaaacac tgttagggaa aattgtgggt
     1801 ggcctttgct gcattgcagg agtcctggtt attgcccttc ctatccccat cattgtgaac
     1861 aatttctctg agttctataa ggaacaaaaa cgccaggaga aagctattaa aaggagagag
     1921 gctcttgaaa gagccaaaag aaatggaagc atcgtttcta tgaacttaaa ggatgccttt
     1981 gctagaagta tggaactgat cgacgtggcc gtggagaagg ctggagagtc ggccaatatc
     2041 aaggactccg tggatgataa ccacctgtct ccaagccggt ggaagtgggc cagaaaggcg
     2101 ctgtcggaaa caagctccaa caagtcttac gagaataagt accaggaggt tagccaaaaa
     2161 gactcccacg aacagctgaa caacacttct tcctccagcc cacagcatct gagcgcccag
     2221 aagctggaga tgctatacaa tgaaatcacc aagacacaga ctcattccca cccgaaccca
     2281 gactgccaag aacagccaga gaggccatct gcctatgagg aggagataga aatggaagag
     2341 gtggtctgcc cacaggagca gctggctgtg gcacagaccg aggtcatcgt ggacatgaag
     2401 agcacctcca gcattgacag cttcaccagc tgtgccactg acttcacgga gactgagagg
     2461 tcacccctgc caccaccctc tgcctctcac ttgcagatga agttccccac cgacctccca
     2521 ggaatggatg agcaccaaag agtcagggca cctccattcc taacactaag cagagataag
     2581 gggcctgctg ccagggaggc tgcactggat tatgccccaa ttgacataac tgtgaacctc
     2641 gatgctgggg cttcccatgg tcctttgcaa cctgacagtg ccagcgacag ccccaagagc
     2701 tcgctgaaag ggagcaaccc cctcaagtct agatccctaa aagtgaactt tcaggaaaac
     2761 agaggcagtg caccccagac cccacccagc acagccaggc cactgccagt aaccacagct
     2821 gactttccgc tcaccacccc ccagcacatg agtaccattc ttctagaaga agccctcccc
     2881 cagggacaga gacccttgcc gggggctgat gtttcagcac actgtcaggg accatccaaa
     2941 ggtctatccc cacgagttcc caagcaaaaa ctgtttcctt tctcttctag agaaaggagg
     3001 agcttcactg aaatagacac tggagaagac gaagacttct tggacctcca aaggccaaga
     3061 ccagacaagc aagcagatcc cagccccaac tgcttagcag ataagcctgg agaggccaga
     3121 gaccctttaa gagaagaggg ctgtgtgggt tcctcctctc cccagaacac agaccacaac
     3181 tgtaggcaag acatttacca ggctgtgggt gaagtcaaaa aggacagtag tcaagaaggg
     3241 tacaagatgg agaaccactt gtttgcccca gaaattcatt ctaacccagg agacacaggt
     3301 tattgcccca ctcgtgagac cagcatgtga ggtaccgcgg gcccgggatc caccggatct
     3361 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     3421 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3481 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3541 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     3601 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3661 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3721 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3781 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3841 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3901 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3961 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     4021 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     4081 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4141 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     4201 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4261 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4321 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4381 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4441 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4501 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     4561 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4621 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4681 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4741 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     4801 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4861 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4921 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4981 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5041 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5101 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5161 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5221 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5281 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5341 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5401 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5461 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5521 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     5581 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     5641 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     5701 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5761 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5821 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5881 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5941 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6001 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6061 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6121 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6181 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6241 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6301 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6361 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6421 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6481 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6541 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6601 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6661 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     6721 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     6781 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     6841 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     6901 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     6961 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     7021 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     7081 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     7141 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     7201 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     7261 agtgaaccgt cagatccgct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag
     7321 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     7381 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     7441 atctgcacca ccggc
//
