LOCUS       sequence-211625-        7301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human KCNB1 C73W in mammalian cells for imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7301)
  TITLE     KCNB1 unpublished plasmids
REFERENCE   2  (bases 1 to 7301)
  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: hC73W-pEGFP-N1
FEATURES             Location/Qualifiers
     source          1..7301
                     /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      2877..2886
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2883..2885,2886..2888,2889..3602)
                     /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(2883..2885,2886..2888,2889..3599)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2928..2949)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3189..3208)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3536..3557
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3725..3846
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3762..3781)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3816..3835
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3853..4308)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3990..4011)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4202..4221
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4335..4439
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4441..4798
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4441..4632
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4464..4484)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4649..4784
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4833..5627
                     /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             4833..5627
                     /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(4887..4906)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5497..5516
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5815..5834)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5859..5906
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6235..6823
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6724..6743
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6998..7301
                     /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 actcagatct cgagctcaag cttcgaattc aaatgccggc gggcatgacg aagcatggct
      301 cccgctccac cagctcgctg ccgcccgagc ccatggagat cgtgcgcagc aaggcgtgct
      361 ctcggcgggt ccgcctcaac gtcggggggc tggcgcacga ggtactctgg cgtaccctgg
      421 accgcctgcc ccgcacgcgg ctgggcaagc tccgcgactg caacacgcac gactcgctgc
      481 tcgaggtgtg ggatgactac agcctcgacg acaacgagta cttctttgac cgccacccgg
      541 gcgccttcac ctccatcctc aacttctacc gcactgggcg actgcacatg atggaggaga
      601 tgtgcgcgct cagcttcagc caagagctcg actactgggg catcgacgag atctacccgg
      661 agtcctgctg ccaggcccgc taccaccaga agaaagagca gatgaacgag gagctcaagc
      721 gtgaggccga gaccctacgg gagcgggaag gcgaggagtt cgataacacg tgctgcgcag
      781 agaagaggaa aaaactctgg gacctactgg agaagcccaa ttcctctgtg gctgccaaga
      841 tccttgccat aatttccatc atgttcatcg tcctctccac cattgccctg tccctcaaca
      901 cgctgcctga gctacagagc ctcgatgagt tcggccagtc cacagacaac ccccagctgg
      961 cccacgtgga ggccgtgtgc atcgcatggt tcaccatgga gtacctgctg aggttcctct
     1021 cctcgcccaa gaagtggaag ttcttcaagg gcccactcaa tgccattgac ttgttggcca
     1081 ttctgccata ctatgtcacc attttcctca ccgaatccaa caagagcgtg ctgcaattcc
     1141 agaatgtccg ccgcgtggtc cagatcttcc gcatcatgcg cattctccgc atccttaagc
     1201 ttgcacgcca ctccactggc ctccagtctc tgggcttcac tttgcggagg agctacaatg
     1261 agttgggctt gctcatcctc ttccttgcca tgggcattat gatcttctcc agccttgtct
     1321 tctttgctga gaaggatgag gacgacacca agttcaaaag catcccagcc tctttctggt
     1381 gggccaccat caccatgact actgttgggt atggagacat ctaccccaag actctcctgg
     1441 ggaaaattgt tgggggactc tgctgcattg caggagtcct ggtgattgct cttcccatcc
     1501 ccatcatcgt caataacttc tctgagttct ataaggagca gaagagacag gagaaagcaa
     1561 tcaaacggcg agaggctctg gagagagcca agaggaatgg cagcatcgta tccatgaaca
     1621 tgaaggatgc ttttgcccgg agcattgaga tgatggacat tgtggttgag aaaaatgggg
     1681 agaatatggg taagaaagac aaagtacaag ataaccactt gtctcctaac aaatggaaat
     1741 ggacaaagag gacactgtct gaaaccagct caagtaagtc ctttgaaacc aaggaacagg
     1801 gatcccctga aaaagccaga tcgtcttcta gtcctcagca cctgaacgtt cagcagttgg
     1861 aagacatgta caataagatg gccaagaccc aatcccaacc catcctcaat accaaggagt
     1921 cagcagcaca gagcaaacca aaggaagaac ttgaaatgga gagtatcccc agccccgtag
     1981 cccctctgcc cactcgcaca gaaggggtca ttgacatgcg aagtatgtca agcattgata
     2041 gtttcattag ctgtgccaca gacttccctg aggccaccag attctcccac agccctttga
     2101 catcactccc cagcaagact gggggcagca cagccccaga agtgggctgg cggggagctc
     2161 tgggtgccag tggtggtagg tttgtggagg ccaaccccag ccctgatgcc agccagcact
     2221 ctagtttctt catcgagagc cccaagagtt ccatgaaaac taacaaccct ttgaagctcc
     2281 gagcacttaa agtcaacttc atggagggtg accccagtcc actcctcccc gttctaggga
     2341 tgtaccatga ccctctcagg aaccggggga gtgctgcggc tgctgtcgct ggactggagt
     2401 gtgccacgct tttggacaag gctgtgctga gcccagagtc ctccatctac accacagcaa
     2461 gtgctaagac acccccccgg tctcctgaga aacacacagc aatagcgttc aactttgagg
     2521 cgggtgtcca ccagtacatt gacgcagaca cagatgatga gggacagctg ctctacagtg
     2581 tggactccag cccccccaaa agcctccctg ggagcaccag tccgaagttc agcacgggga
     2641 caagatcgga gaaaaaccac tttgaaagct cccctttacc cacctcccct aagttcttaa
     2701 ggcagaactg tatttactcc acagaagcat tgactggaaa aggccccagt ggtcaggaaa
     2761 agtgcaaact tgagaaccac atctcccctg acgtccgtgt gttgccaggg ggaggagccc
     2821 atggaagcac acgagatcag agcatcacgg taccgcgggc ccgggatcca ccggtcgcca
     2881 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     2941 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     3001 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     3061 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     3121 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     3181 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     3241 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     3301 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     3361 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     3421 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     3481 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     3541 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     3601 aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3661 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3721 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3781 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3841 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3901 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3961 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4021 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4081 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4141 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     4201 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     4261 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     4321 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4381 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     4441 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4501 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4561 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4621 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4681 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4741 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     4801 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4861 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4921 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4981 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     5041 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5101 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5161 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5221 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5281 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5341 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5401 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5461 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5521 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5581 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5641 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5701 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5761 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     5821 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     5881 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     5941 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     6001 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     6061 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     6121 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6181 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6241 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6301 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6361 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6421 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6481 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6541 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6601 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6661 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6721 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6781 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6841 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6901 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
     6961 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     7021 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     7081 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     7141 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     7201 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     7261 cttggcagta catctacgta ttagtcatcg ctattaccat g
//
