LOCUS       sequence-211650-        8063 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human KCNB1 C73L in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8063)
  TITLE     KCNB1 unpublished plasmids
REFERENCE   2  (bases 1 to 8063)
  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: hC73L-HA
FEATURES             Location/Qualifiers
     source          1..8063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     90..110
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     136..160
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          300..432
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     415..434
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        477..495
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3101..3127
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3177..3298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3188..3207)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3242..3261
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3484..3939
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3571..3590)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3781..3802
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4056..4413
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4056..4247
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4079..4099)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4264..4399
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4464..5258
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4518..4537)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5128..5147
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5322..5370
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5411..5430)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5530..5552
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5590..5608)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5676..5780
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5781..5849,5850..6641)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5999..6018)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6812..7400
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7301..7320
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7611..7990
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7991..8063,1..139)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
       61 gtaacaactg cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga
      121 ggtctatata agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt
      181 agtttatcac agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt
      241 aagctgcagt gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg
      301 taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac
      361 agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct
      421 ttctctccac aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat
      481 acgactcact ataggctagc ctcgagagct gaattccacc atgccggcgg gcatgacgaa
      541 gcatggctcc cgctccacca gctcgctgcc gcccgagccc atggagatcg tgcgcagcaa
      601 ggcgtgctct cggcgggtcc gcctcaacgt cggggggctg gcgcacgagg tactctggcg
      661 taccctggac cgcctgcccc gcacgcggct gggcaagctc cgcgactgca acacgcacga
      721 ctcgctgctc gaggtgctcg atgactacag cctcgacgac aacgagtact tctttgaccg
      781 ccacccgggc gccttcacct ccatcctcaa cttctaccgc actgggcgac tgcacatgat
      841 ggaggagatg tgcgcgctca gcttcagcca agagctcgac tactggggca tcgacgagat
      901 ctacccggag tcctgctgcc aggcccgcta ccaccagaag aaagagcaga tgaacgagga
      961 gctcaagcgt gaggccgaga ccctacggga gcgggaaggc gaggagttcg ataacacgtg
     1021 ctgcgcagag aagaggaaaa aactctggga cctactggag aagcccaatt cctctgtggc
     1081 tgccaagatc cttgccataa tttccatcat gttcatcgtc ctctccacca ttgccctgtc
     1141 cctcaacacg ctgcctgagc tacagagcct cgatgagttc ggccagtcca cagacaaccc
     1201 ccagctggcc cacgtggagg ccgtgtgcat cgcatggttc accatggagt acctgctgag
     1261 gttcctctcc tcgcccaaga agtggaagtt cttcaagggc ccactcaatg ccattgactt
     1321 gttggccatt ctgccatact atgtcaccat tttcctcacc gaatccaaca agagcgtgct
     1381 gcaattccag aatgtccgcc gcgtggtcca gatcttccgc atcatgcgca ttctccgcat
     1441 ccttaagctt gcacgccact ccactggcct ccagtctctg ggcttcactt tgcggaggag
     1501 ctacaatgag ttgggcttgc tcatcctctt ccttgccatg ggcattatga tcttctccag
     1561 ccttgtcttc tttgctgaga aggatgagga cgacaccaag ttcaaaagca tcccagcctc
     1621 tttctggtgg gccaccatca ccatgactac tgttgggtat ggagacatct accccaagac
     1681 tctcctgggg aaaattgttg ggggactctg ctgcattgca ggagtcctgg tgattgctct
     1741 tcccatcccc atcatcgtca ataacttctc tgagttctat aaggagcaga agagacagga
     1801 gaaagcaatc aaacggcgag aggctctgga gagagccaag aggaatggca gcatcgtatc
     1861 catgaacatg aaggatgctt ttgcccggag cattgagatg atggacattg tggttgagaa
     1921 aaatggggag aatatgggta agaaagacaa agtacaagat aaccacttgt ctcctaacaa
     1981 atggaaatgg acaaagagga cactgtctga aaccagctca agtaagtcct ttgaaaccaa
     2041 ggaacaggga tcccctgaaa aagccagatc gtcttctagt cctcagcacc tgaacgttca
     2101 gcagttggaa gacatgtaca ataagatggc caagacccaa tcccaaccca tcctcaatac
     2161 caaggagtca gcagcacaga gcaaaccaaa ggaagaactt gaaatggaga gtatccccag
     2221 ccccgtagcc cctctgccca ctcgcacaga aggggtcatt gacatgcgaa gtatgtcaag
     2281 cattgatagt ttcattagct gtgccacaga cttccctgag gccaccagat tctcccacag
     2341 ccctttgaca tcactcccca gcaagactgg gggcagcaca gccccagaag tgggctggcg
     2401 gggagctctg ggtgccagtg gtggtaggtt tgtggaggcc aaccccagcc ctgatgccag
     2461 ccagcactct agtttcttca tcgagagccc caagagttcc atgaaaacta acaacccttt
     2521 gaagctccga gcacttaaag tcaacttcat ggagggtgac cccagtccac tcctccccgt
     2581 tctagggatg taccatgacc ctctcaggaa ccgggggagt gctgcggctg ctgtcgctgg
     2641 actggagtgt gccacgcttt tggacaaggc tgtgctgagc ccagagtcct ccatctacac
     2701 cacagcaagt gctaagacac ccccccggtc tcctgagaaa cacacagcaa tagcgttcaa
     2761 ctttgaggcg ggtgtccacc agtacattga cgcagacaca gatgatgagg gacagctgct
     2821 ctacagtgtg gactccagcc cccccaaaag cctccctggg agcaccagtc cgaagttcag
     2881 cacggggaca agatcggaga aaaaccactt tgaaagctcc cctttaccca cctcccctaa
     2941 gttcttaagg cagaactgta tttactccac agaagcattg actggaaaag gccccagtgg
     3001 tcaggaaaag tgcaaacttg agaaccacat ctcccctgac gtccgtgtgt tgccaggggg
     3061 aggagcccat ggaagcacac gagatcagag catcgaattc tacccatacg atgttccaga
     3121 ttacgcttag gcggccgctt ccctttagtg agggttaatg cttcgagcag acatgataag
     3181 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     3241 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3301 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta
     3361 aagcaagtaa aacctctaca aatgtggtaa aatccgataa ggatcgatcc gggctggcgt
     3421 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     3481 tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3541 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3601 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3661 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3721 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3781 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3841 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3901 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct
     3961 tacgcatctg tgcggtattt cacaccgcat acgcggatct gcgcagcacc atggcctgaa
     4021 ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg
     4081 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4141 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4201 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4261 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4321 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4381 ggaggctttt ttggaggcct aggcttttgc aaaaagcttg attcttctga cacaacagtc
     4441 tcgaacttaa ggctagagcc accatgattg aacaagatgg attgcacgca ggttctccgg
     4501 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4561 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4621 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4681 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4741 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4801 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4861 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4921 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4981 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5041 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5101 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5161 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5221 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5281 gaccgaccaa gcgacgccca acctgccatc acgatggccg caataaaata tctttatttt
     5341 cattacatct gtgtgttggt tttttgtgtg aatcgatagc gataaggatc cgcgtatggt
     5401 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     5461 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     5521 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     5581 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     5641 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     5701 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5761 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     5821 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5881 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5941 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     6001 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6061 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6121 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6181 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6241 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6301 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     6361 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6421 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6481 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6541 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6601 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     6661 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6721 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6781 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6841 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6901 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6961 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7021 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7081 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7141 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7201 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7261 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7321 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7381 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7441 gctggccttt tgctcacatg gctcgacaga tcttcaatat tggccattag ccatattatt
     7501 cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata
     7561 tcataatatg tacatttata ttggctcatg tccaatatga ccgccatgtt ggcattgatt
     7621 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     7681 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     7741 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     7801 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     7861 tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     7921 ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     7981 tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc
     8041 acggggattt ccaagtctcc acc
//
