LOCUS       sequence-217177-        7299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human KCNB1 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  TITLE     KCNB1 unpublished plasmids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: hKv2.1-pEGFP-C1
FEATURES             Location/Qualifiers
     source          1..7299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      39..48
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(45..47,48..50,51..761)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(45..47,48..50,51..761)
                     /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(90..111)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(351..370)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     698..719
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3519..3640
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3556..3575)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3610..3629
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3647..4102)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3784..3805)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3996..4015
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4129..4233
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4235..4592
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4235..4426
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4258..4278)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4443..4578
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4627..5421
                     /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             4627..5421
                     /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(4681..4700)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5291..5310
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5609..5628)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5653..5700
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6029..6617
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6518..6537
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6792..7095
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7096..7299
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7250..7270
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ggtttagtga accgtcagat ccgctagcgc taccggtcgc caccatggtg agcaagggcg
       61 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      121 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      181 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
      241 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
      301 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
      361 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
      421 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
      481 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
      541 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
      601 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
      661 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
      721 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtccggactc agatctcgag
      781 ctcaagcttc gaattcaaat gccggcgggc atgacgaagc atggctcccg ctccaccagc
      841 tcgctgccgc ccgagcccat ggagatcgtg cgcagcaagg cgtgctctcg gcgggtccgc
      901 ctcaacgtcg gggggctggc gcacgaggta ctctggcgta ccctggaccg cctgccccgc
      961 acgcggctgg gcaagctccg cgactgcaac acgcacgact cgctgctcga ggtgtgcgat
     1021 gactacagcc tcgacgacaa cgagtacttc tttgaccgcc acccgggcgc cttcacctcc
     1081 atcctcaact tctaccgcac tgggcgactg cacatgatgg aggagatgtg cgcgctcagc
     1141 ttcagccaag agctcgacta ctggggcatc gacgagatct acccggagtc ctgctgccag
     1201 gcccgctacc accagaagaa agagcagatg aacgaggagc tcaagcgtga ggccgagacc
     1261 ctacgggagc gggaaggcga ggagttcgat aacacgtgct gcgcagagaa gaggaaaaaa
     1321 ctctgggacc tactggagaa gcccaattcc tctgtggctg ccaagatcct tgccataatt
     1381 tccatcatgt tcatcgtcct ctccaccatt gccctgtccc tcaacacgct gcctgagcta
     1441 cagagcctcg atgagttcgg ccagtccaca gacaaccccc agctggccca cgtggaggcc
     1501 gtgtgcatcg catggttcac catggagtac ctgctgaggt tcctctcctc gcccaagaag
     1561 tggaagttct tcaagggccc actcaatgcc attgacttgt tggccattct gccatactat
     1621 gtcaccattt tcctcaccga atccaacaag agcgtgctgc aattccagaa tgtccgccgc
     1681 gtggtccaga tcttccgcat catgcgcatt ctccgcatcc ttaagcttgc acgccactcc
     1741 actggcctcc agtctctggg cttcactttg cggaggagct acaatgagtt gggcttgctc
     1801 atcctcttcc ttgccatggg cattatgatc ttctccagcc ttgtcttctt tgctgagaag
     1861 gatgaggacg acaccaagtt caaaagcatc ccagcctctt tctggtgggc caccatcacc
     1921 atgactactg ttgggtatgg agacatctac cccaagactc tcctggggaa aattgttggg
     1981 ggactctgct gcattgcagg agtcctggtg attgctcttc ccatccccat catcgtcaat
     2041 aacttctctg agttctataa ggagcagaag agacaggaga aagcaatcaa acggcgagag
     2101 gctctggaga gagccaagag gaatggcagc atcgtatcca tgaacatgaa ggatgctttt
     2161 gcccggagca ttgagatgat ggacattgtg gttgagaaaa atggggagaa tatgggtaag
     2221 aaagacaaag tacaagataa ccacttgtct cctaacaaat ggaaatggac aaagaggaca
     2281 ctgtctgaaa ccagctcaag taagtccttt gaaaccaagg aacagggatc ccctgaaaaa
     2341 gccagatcgt cttctagtcc tcagcacctg aacgttcagc agttggaaga catgtacaat
     2401 aagatggcca agacccaatc ccaacccatc ctcaatacca aggagtcagc agcacagagc
     2461 aaaccaaagg aagaacttga aatggagagt atccccagcc ccgtagcccc tctgcccact
     2521 cgcacagaag gggtcattga catgcgaagt atgtcaagca ttgatagttt cattagctgt
     2581 gccacagact tccctgaggc caccagattc tcccacagcc ctttgacatc actccccagc
     2641 aagactgggg gcagcacagc cccagaagtg ggctggcggg gagctctggg tgccagtggt
     2701 ggtaggtttg tggaggccaa ccccagccct gatgccagcc agcactctag tttcttcatc
     2761 gagagcccca agagttccat gaaaactaac aaccctttga agctccgagc acttaaagtc
     2821 aacttcatgg agggtgaccc cagtccactc ctccccgttc tagggatgta ccatgaccct
     2881 ctcaggaacc gggggagtgc tgcggctgct gtcgctggac tggagtgtgc cacgcttttg
     2941 gacaaggctg tgctgagccc agagtcctcc atctacacca cagcaagtgc taagacaccc
     3001 ccccggtctc ctgagaaaca cacagcaata gcgttcaact ttgaggcggg tgtccaccag
     3061 tacattgacg cagacacaga tgatgaggga cagctgctct acagtgtgga ctccagcccc
     3121 cccaaaagcc tccctgggag caccagtccg aagttcagca cggggacaag atcggagaaa
     3181 aaccactttg aaagctcccc tttacccacc tcccctaagt tcttaaggca gaactgtatt
     3241 tactccacag aagcattgac tggaaaaggc cccagtggtc aggaaaagtg caaacttgag
     3301 aaccacatct cccctgacgt ccgtgtgttg ccagggggag gagcccatgg aagcacacga
     3361 gatcagagca tcacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa
     3421 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3481 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3541 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3601 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     3661 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3721 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3781 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3841 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3901 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3961 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4021 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4081 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4141 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4201 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     4261 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4321 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4381 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4441 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4501 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     4561 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     4621 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4681 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4741 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4801 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4861 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4921 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4981 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5041 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5101 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     5161 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5221 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5281 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5341 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5401 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5461 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5521 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5581 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     5641 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     5701 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     5761 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     5821 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     5881 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5941 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6001 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6061 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6121 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6181 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6241 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6301 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6361 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6421 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6481 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6541 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6601 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6661 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6721 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
     6781 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     6841 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     6901 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     6961 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     7021 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     7081 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     7141 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     7201 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     7261 ggtaggcgtg tacggtggga ggtctatata agcagagct
//
