LOCUS       sequence-217377-        6900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses worm KHT-1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6900)
  AUTHORS   Cai SQ, Wang Y, Park KH, Tong X, Pan Z, Sesti F
  TITLE     Auto-phosphorylation of a voltage-gated K+ channel controls 
            non-associative learning.
  JOURNAL   EMBO J. 2009 Jun 3;28(11):1601-11. doi: 10.1038/emboj.2009.112. Epub
            2009 Apr 23.
  PUBMED    19387491
REFERENCE   2  (bases 1 to 6900)
  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"; doi: 
            "10.1038/emboj.2009.112"; journalName: "EMBO J"; date: "2009-06-3- 
            3"; volume: "28"; issue: "11"; pages: "1601-11"
COMMENT     Sequence Label: KHT-1
FEATURES             Location/Qualifiers
     source          1..6900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     589..613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          753..885
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     868..887
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        930..948
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2467..2588
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2478..2497)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2532..2551
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2774..3229
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2861..2880)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3071..3092
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3346..3703
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3346..3537
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3369..3389)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3554..3689
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3754..4548
                     /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(3808..3827)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4418..4437
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4612..4660
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4701..4720)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4820..4842
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4880..4898)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4966..5070
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5071..5139,5140..5931)
                     /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(5289..5308)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6102..6690
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6591..6610
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctgcgatcgc ccgccccgtt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
      601 gaaccgtcag atcactagaa gctttattgc ggtagtttat cacagttaaa ttgctaacgc
      661 agtcagtgct tctgacacaa cagtctcgaa cttaagctgc agtgactctc ttaaggtagc
      721 cttgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt
      781 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
      841 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt
      901 tcaattacag ctcttaaggc tagagtactt aatacgactc actataggct agcctcgaga
      961 tggacagtga ataccggatc atactcaacg tgggtggtgt tcgacacgaa acctataagc
     1021 atacgcttaa gaaaattcca gctacacgac tatcccggtt gactacaaat ttggccaatt
     1081 atgatccagt tttaaacgag tattttttcg atagacatcc cggagttttt gcgcaaatac
     1141 taaattatta tagaacgggc aaattgcatt atcctcttga cgtgtgtgga ccattatttg
     1201 aagaggagtt aaagtattgg ggacttgatt caaatgaatg cgaaccatgc tgttggatga
     1261 cattatctgg tacaagagat acacaagatg tgctcaaaac attagacgag ttggatattg
     1321 aagttgatca aacagaagga cctgaactgt acaaacgatt tggttgggag gatgactatc
     1381 ataatgatag tctcagtgcc tggcagaaga tcaaacctcg gatttggagg cttttcgatg
     1441 aaccaaattc atctagatca gcacaattta tagcagctat ttctgtattt tttctgataa
     1501 cagccatcat tgttttctgt ctgaaaaccc atcctggttt gaggatccct gaattagctc
     1561 catttggaaa cttttcgcgg aatcatcgga gtacatcttc acgaatacat cctgcacaga
     1621 taaacattga taaggccaat tctaggcctc atccaacttt tatgtacata gaaacaatat
     1681 gtaacatttg gttcacgatt gaaattctgg caagattctc ttcatgtcca tctcgttttg
     1741 aatatcttcg tgcacctgtg aacattatcg acattgttgc cactttgaca ttctacattg
     1801 atttattatc aagcatgttt ggagcaactg cagatcttga gttcttctcc atcattcgta
     1861 ttatgagact tttcaaattg actcatcata attcgggact caagattttg atgcacacct
     1921 tccgagcatc agcaaaggaa ttgatgctac ttgtattctt tttggtattg ggagttgttg
     1981 tatttgcatc actggtgtat tatgcagaac gggtggaaag taatgaagat aatcaatttg
     2041 tgtcaattcc gattggattg tggtgggcga tcgtcacaat gacaacaatt ggctatggtg
     2101 acataacacc tcatacatat ctgggccgcc tgattggttc gatttgtgct ctcgctggtg
     2161 tattgacaat cgctttgcca gttccagtca tcgtctctaa cttcgccatg ttctattccc
     2221 atacacaagc tcgttcaaag atgcccaaga aaagacgagg agtgctctca atggatcaag
     2281 ttgctcagca tgtaaaacat gctccggttc gaaattccaa tggaccaacg caacggaaga
     2341 atatggaaaa tacaccactt gtatccaaca accatacacc gccacatacc ttaatctagt
     2401 tgtttcatag gtcgacccgg gcggccgctt ccctttagtg agggttaatg cttcgagcag
     2461 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     2521 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     2581 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     2641 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatccgataa ggatcgatcc
     2701 gggctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     2761 gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2821 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2881 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2941 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3001 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3061 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3121 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3181 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt
     3241 attttctcct tacgcatctg tgcggtattt cacaccgcat acgcggatct gcgcagcacc
     3301 atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa
     3361 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3421 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3481 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3541 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3601 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3661 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg attcttctga
     3721 cacaacagtc tcgaacttaa ggctagagcc accatgattg aacaagatgg attgcacgca
     3781 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3841 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3901 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     3961 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4021 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4081 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4141 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4201 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4261 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4321 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4381 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4441 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4501 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4561 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgatggccg caataaaata
     4621 tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagc gataaggatc
     4681 cgcgtatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     4741 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4801 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4861 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4921 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4981 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5041 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5101 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5161 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5221 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5281 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5341 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5401 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5461 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5521 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5581 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5641 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5701 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5761 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5821 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5881 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5941 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6001 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6061 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6121 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6181 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6241 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6301 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6361 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6421 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6481 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6541 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6601 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6661 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6721 ctggcctttt gctggccttt tgctcacatg gctcgacaga tcttcaatat tggccattag
     6781 ccatattatt cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt
     6841 tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga ccgccatgtt
//
