LOCUS       sequence-215135-        7797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Electrophysiology in HEK cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7797)
  AUTHORS   Chang A, Abderemane-Ali F, Hura GL, Rossen ND, Gate RE, Minor DL Jr.
  TITLE     A Calmodulin C-Lobe Ca(2+)-Dependent Switch Governs Kv7 Channel 
            Function.
  JOURNAL   Neuron. 2018 Feb 21;97(4):836-852.e6. doi: 
            10.1016/j.neuron.2018.01.035. Epub 2018 Feb 8.
  PUBMED    29429937
REFERENCE   2  (bases 1 to 7797)
  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: 
            "10.1016/j.neuron.2018.01.035"; journalName: "Neuron"; date: 
            "2018-02-21- 21"; volume: "97"; issue: "4"; pages: "836-852.e6"
COMMENT     Sequence Label: Kv7.4 (1-645, Delta-368-492)/pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..7797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2528..2548
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             join(2549..2551,2552..3262)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2549..2551,2552..3262)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2591..2612)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2852..2871)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3199..3220
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3263..3286
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     promoter        complement(3350..3368)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3351..3368)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3388..3405)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3394..3618
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3664..4092
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3751..3770)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3961..3982
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4101..4121)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4106..4435
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4286..4421
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4502..5296
                     /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(4556..4575)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5166..5185
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5470..5591
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5507..5526)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5561..5580
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5640..5656)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5640..5656)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5653..5675)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5664..5680
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5688..5694,5695..5712,5713..5718))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5733..5754
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5871..5888)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6042..6630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6122..6141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6801..7592,7593..7661))
                     /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     7424..7443
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7662..7766)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccttgggtac ttccaatcca tttgatggcc gaggcccccc cgcgccgcct
      961 cggcctgggt cccccgcccg gggacgcccc ccgcgcggag ctagtggcgc tcacggccgt
     1021 gcagagcgaa cagggcgagg cgggcggggg cggctccccg cgccgcctcg gcctcctggg
     1081 cagccccctg ccgccgggcg cgcccctccc tgggccgggc tccggctcgg gctccgcctg
     1141 cggccagcgc tcctcggccg cgcacaagcg ctaccgccgc ctgcagaact gggtctacaa
     1201 cgtgctggag cggccccgcg gctgggcctt cgtctaccac gtcttcatat ttttgctggt
     1261 cttcagctgt ctggtgctgt ctgtgctgtc cactatccag gagcaccagg aacttgccaa
     1321 cgagtgtctc ctcatcttgg aattcgtgat gatcgtggtt ttcggcttgg agtacatcgt
     1381 ccgggtctgg tccgccggat gctgctgccg ctaccgagga tggcagggtc gcttccgctt
     1441 tgccagaaag cccttctgtg tcatcgactt catcgtgttc gtggcctcgg tggccgtcat
     1501 cgccgcgggt acccagggca acatcttcgc cacgtccgcg ctgcgcagca tgcgcttcct
     1561 gcagatcctg cgcatggtgc gcatggaccg ccgcggcggc acctggaagc tgctgggctc
     1621 agtggtctac gcgcatagca aggagctgat caccgcctgg tacatcgggt tcctggtgct
     1681 catcttcgcc tccttcctgg tctacctggc cgagaaggac gccaactccg acttctcctc
     1741 ctacgccgac tcgctctggt gggggacgat tacattgaca accatcggct atggtgacaa
     1801 gacaccgcac acatggctgg gcagggtcct ggctgctggc ttcgccttac tgggcatctc
     1861 tttctttgcc ctgcctgccg gcatcctagg ctccggcttt gccctgaagg tccaggagca
     1921 gcaccggcag aagcacttcg agaagcggag gatgccggca gccaacctca tccaggctgc
     1981 ctggcgcctg tactccaccg atatgagccg ggcctacctg acagccacct ggtacaagct
     2041 tgcatctctg agactcaaac cccgcacctc tgctgaggat gccccctcag aggaagtagc
     2101 agaggagaag agctaccagt gtgagctcac ggtggacgac atcatgcctg ctgtgaagac
     2161 agtcatccgc tccatcagga ttctcaagtt cctggtggcc aaaaggaaat tcaaggagac
     2221 actgcgaccg tacgacgtga aggacgtcat tgagcagtac tcagcaggcc acctggacat
     2281 gctgggccgg atcaagagcc tgcaaactcg ggtggaccaa attgtgggtc gggggcccgg
     2341 ggacaggaag gcccgggaga agggcgacaa ggggccctcc gacgcggagg tggtggatga
     2401 aatcagcatg atgggacgcg tggtcaaggt ggagaagcag gtgcagtcca tcgagcacaa
     2461 gctggacctg ctgttgggct tctattcgcg cgccctgcgc taaggcaaat tgggattaat
     2521 taatctcgaa aacctgtact tccaatccgt gagcaagggc gaggagctgt tcaccggggt
     2581 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     2641 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     2701 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     2761 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     2821 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     2881 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     2941 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     3001 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     3061 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     3121 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     3181 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     3241 cggcatggac gagctgtaca agcatcacca tcaccatcac catcactaag gatcctgcag
     3301 atatccatca cactggcggc cgctcgagca tgcatctaga gggccctatt ctatagtgtc
     3361 acctaaatgc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     3421 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     3481 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     3541 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     3601 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     3661 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3721 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3781 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3841 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3901 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3961 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4021 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4081 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4141 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4201 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4261 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4321 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     4381 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4441 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     4501 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4561 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4621 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4681 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4741 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4801 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4861 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4921 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4981 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     5041 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5101 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5161 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5221 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5281 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5341 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5401 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5461 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5521 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5581 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5641 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5701 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5761 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5821 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5881 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5941 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6001 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6061 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6121 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6181 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6241 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6301 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6361 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6421 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6481 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6541 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6601 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6661 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6721 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6781 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6841 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6901 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6961 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7021 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7081 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7141 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7201 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7261 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7321 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7381 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7441 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7501 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7561 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7621 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7681 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7741 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
