LOCUS       sequence-217175-        7299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human KCNB1 C73S in mammalian cells for imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7299)
  TITLE     KCNB1 unpublished plasmids
REFERENCE   2  (bases 1 to 7299)
  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: hC73S-pEGFP-C1
FEATURES             Location/Qualifiers
     source          1..7299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      243..252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(249..251,252..254,255..965)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(249..251,252..254,255..965)
                     /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(294..315)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(555..574)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     902..923
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3723..3844
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3760..3779)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3814..3833
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3851..4306)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3988..4009)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4200..4219
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4333..4437
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4439..4796
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4439..4630
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4462..4482)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4647..4782
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4831..5625
                     /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             4831..5625
                     /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(4885..4904)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5495..5514
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5813..5832)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5857..5904
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6233..6821
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6722..6741
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6996..7299
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      301 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      361 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      421 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
      481 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      541 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      601 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      661 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
      721 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      781 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      841 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
      901 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
      961 acaagtccgg actcagatct cgagctcaag cttcgaattc aaatgccggc gggcatgacg
     1021 aagcatggct cccgctccac cagctcgctg ccgcccgagc ccatggagat cgtgcgcagc
     1081 aaggcgtgct ctcggcgggt ccgcctcaac gtcggggggc tggcgcacga ggtactctgg
     1141 cgtaccctgg accgcctgcc ccgcacgcgg ctgggcaagc tccgcgactg caacacgcac
     1201 gactcgctgc tcgaggtgag cgatgactac agcctcgacg acaacgagta cttctttgac
     1261 cgccacccgg gcgccttcac ctccatcctc aacttctacc gcactgggcg actgcacatg
     1321 atggaggaga tgtgcgcgct cagcttcagc caagagctcg actactgggg catcgacgag
     1381 atctacccgg agtcctgctg ccaggcccgc taccaccaga agaaagagca gatgaacgag
     1441 gagctcaagc gtgaggccga gaccctacgg gagcgggaag gcgaggagtt cgataacacg
     1501 tgctgcgcag agaagaggaa aaaactctgg gacctactgg agaagcccaa ttcctctgtg
     1561 gctgccaaga tccttgccat aatttccatc atgttcatcg tcctctccac cattgccctg
     1621 tccctcaaca cgctgcctga gctacagagc ctcgatgagt tcggccagtc cacagacaac
     1681 ccccagctgg cccacgtgga ggccgtgtgc atcgcatggt tcaccatgga gtacctgctg
     1741 aggttcctct cctcgcccaa gaagtggaag ttcttcaagg gcccactcaa tgccattgac
     1801 ttgttggcca ttctgccata ctatgtcacc attttcctca ccgaatccaa caagagcgtg
     1861 ctgcaattcc agaatgtccg ccgcgtggtc cagatcttcc gcatcatgcg cattctccgc
     1921 atccttaagc ttgcacgcca ctccactggc ctccagtctc tgggcttcac tttgcggagg
     1981 agctacaatg agttgggctt gctcatcctc ttccttgcca tgggcattat gatcttctcc
     2041 agccttgtct tctttgctga gaaggatgag gacgacacca agttcaaaag catcccagcc
     2101 tctttctggt gggccaccat caccatgact actgttgggt atggagacat ctaccccaag
     2161 actctcctgg ggaaaattgt tgggggactc tgctgcattg caggagtcct ggtgattgct
     2221 cttcccatcc ccatcatcgt caataacttc tctgagttct ataaggagca gaagagacag
     2281 gagaaagcaa tcaaacggcg agaggctctg gagagagcca agaggaatgg cagcatcgta
     2341 tccatgaaca tgaaggatgc ttttgcccgg agcattgaga tgatggacat tgtggttgag
     2401 aaaaatgggg agaatatggg taagaaagac aaagtacaag ataaccactt gtctcctaac
     2461 aaatggaaat ggacaaagag gacactgtct gaaaccagct caagtaagtc ctttgaaacc
     2521 aaggaacagg gatcccctga aaaagccaga tcgtcttcta gtcctcagca cctgaacgtt
     2581 cagcagttgg aagacatgta caataagatg gccaagaccc aatcccaacc catcctcaat
     2641 accaaggagt cagcagcaca gagcaaacca aaggaagaac ttgaaatgga gagtatcccc
     2701 agccccgtag cccctctgcc cactcgcaca gaaggggtca ttgacatgcg aagtatgtca
     2761 agcattgata gtttcattag ctgtgccaca gacttccctg aggccaccag attctcccac
     2821 agccctttga catcactccc cagcaagact gggggcagca cagccccaga agtgggctgg
     2881 cggggagctc tgggtgccag tggtggtagg tttgtggagg ccaaccccag ccctgatgcc
     2941 agccagcact ctagtttctt catcgagagc cccaagagtt ccatgaaaac taacaaccct
     3001 ttgaagctcc gagcacttaa agtcaacttc atggagggtg accccagtcc actcctcccc
     3061 gttctaggga tgtaccatga ccctctcagg aaccggggga gtgctgcggc tgctgtcgct
     3121 ggactggagt gtgccacgct tttggacaag gctgtgctga gcccagagtc ctccatctac
     3181 accacagcaa gtgctaagac acccccccgg tctcctgaga aacacacagc aatagcgttc
     3241 aactttgagg cgggtgtcca ccagtacatt gacgcagaca cagatgatga gggacagctg
     3301 ctctacagtg tggactccag cccccccaaa agcctccctg ggagcaccag tccgaagttc
     3361 agcacgggga caagatcgga gaaaaaccac tttgaaagct cccctttacc cacctcccct
     3421 aagttcttaa ggcagaactg tatttactcc acagaagcat tgactggaaa aggccccagt
     3481 ggtcaggaaa agtgcaaact tgagaaccac atctcccctg acgtccgtgt gttgccaggg
     3541 ggaggagccc atggaagcac acgagatcag agcatcacgg taccgcgggc ccgggatcca
     3601 ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     3661 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3721 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3781 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3841 cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3901 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3961 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     4021 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     4081 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     4141 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     4201 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     4261 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     4321 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4381 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     4441 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4501 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4561 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4621 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4681 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     4741 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     4801 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4861 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4921 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4981 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     5041 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5101 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5161 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5221 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5281 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5341 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5401 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5461 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5521 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5581 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5641 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5701 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5761 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     5821 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     5881 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     5941 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     6001 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     6061 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     6121 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6181 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6241 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6301 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6361 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6421 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6481 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6541 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6601 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6661 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6721 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6781 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6841 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6901 ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg
     6961 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     7021 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     7081 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     7141 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     7201 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     7261 tggcagtaca tctacgtatt agtcatcgct attaccatg
//
