LOCUS       sequence-202201-        9013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses BKa (KCNMA1, mSlo alpha subunit) in mammalian cells. 
            (tagless, leaderless).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9013)
  AUTHORS   Pratt CP, He J, Wang Y, Barth AL, Bruchez MP
  TITLE     Fluorogenic Green-Inside Red-Outside (GIRO) Labeling Approach 
            Reveals Adenylyl Cyclase-Dependent Control of BKalpha Surface 
            Expression.
  JOURNAL   Bioconjug Chem. 2015 Sep 16;26(9):1963-71. doi: 
            10.1021/acs.bioconjchem.5b00409. Epub 2015 Sep 2.
  PUBMED    26301573
REFERENCE   2  (bases 1 to 9013)
  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: 
            "10.1021/acs.bioconjchem.5b00409"; journalName: "Bioconjug Chem"; 
            date: "2015-09-16- 16"; volume: "26"; issue: "9"; pages: "1963-71"
COMMENT     Sequence Label: pcDNA3.1-BKa
FEATURES             Location/Qualifiers
     source          1..9013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        110..491
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        492..695
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     646..666
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        740..758
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4245..4379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4479..4496)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4485..4709
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4755..5183
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4842..4861)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5052..5073
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5192..5212)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5197..5526
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5377..5512
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5439..5458
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5593..6387
                     /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(5647..5666)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6257..6276
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6561..6682
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6731..6747)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6731..6747)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6744..6766)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6755..6771
                     /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(6779..6785,6786..6803,6804..6809))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6824..6845
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6962..6979)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7133..7721)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7213..7232)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7892..8683,8684..8752))
                     /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     8515..8534
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8753..8857)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8932..8951)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
       61 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
      121 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      181 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      241 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      301 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      361 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      421 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      481 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      541 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      601 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      661 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
      721 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct ggctagtatg
      781 gatgcgctca tcataccggt gaccatggag gtgccgtgcg acagccgggg ccaacgcatg
      841 tggtgggctt tcttggcctc ctccatggtg actttcttcg ggggcctctt catcatcttg
      901 ctctggcgga cgctcaagta cctgtggacc gtttgctgcc actgcggggg caagacgaag
      961 gaggcccaga agataaacaa tggctccagc caggcagatg gtactctcaa gccagtggac
     1021 gaaaaagagg aggtggtggc agccgaggtc ggctggatga catctgtgaa ggactgggca
     1081 ggggtgatga tatccgccca gacactgact ggcagagtcc tggttgtgtt agtctttgct
     1141 ctcagcattg gtgccctcgt aatatacttc atagactcgt caaacccaat agaatcctgc
     1201 cagaatttct acaaagattt cacattacag atcgacatgg ctttcaacgt gttcttcctc
     1261 ctctactttg gcttgcggtt tattgcagcc aacgataagc tgtggttctg gctggaagtg
     1321 aattcagtag tagatttctt cacagtccct cctgtgtttg tgtctgtgta cttaaacaga
     1381 agttggcttg gcttgagatt tttaagagct ctcagactga tacagttttc agagattttg
     1441 cagtttctga atatccttaa aacaagtaac tccatcaagc tggtgaatct gctctccata
     1501 tttatcagca cgtggctgac tgcagctgga ttcatccact tggtggagaa ttcaggggac
     1561 ccatgggaaa atttccaaaa caaccaggca cttacgtact gggaatgtgt ctacttactc
     1621 atggtcacaa tgtctacagt gggttatggg gacgtttatg caaaaaccac acttggacgc
     1681 ctcttcatgg tcttcttcat cctcggggga ctggccatgt ttgccagcta cgtccctgaa
     1741 atcatagagt taataggaaa ccgcaagaaa tacgggggct cctatagcgc ggttagtgga
     1801 agaaagcaca ttgtagtctg tggacacatt actctggaga gtgtctctaa cttcctgaag
     1861 gactttctgc acaaggaccg ggatgatgtc aacgtggaga ttgtctttct tcacaacatc
     1921 tcccctaacc ttgagcttga ggctctgttc aaacggcatt tcactcaggt ggagttttat
     1981 cagggctctg tcctcaatcc acatgatctt gccagagtca agatagagtc agcagatgca
     2041 tgcctgatcc ttgccaataa gtattgcgct gacccggatg cagaagatgc ctccaacatc
     2101 atgagagtga tctccatcaa aaactaccac ccaaagatca ggatcatcac tcagatgctg
     2161 cagtatcaca acaaggccca tctgctcaac atccccagct ggaactggaa agagggtgat
     2221 gacgcaatat gccttgcaga gctcaagttg ggtttcatag cccagagctg tctggctcaa
     2281 ggcctctcca caatgcttgc caatctcttc tctatgaggt cattcataaa gattgaggaa
     2341 gacacatggc agaaatacta cttggaagga gtctccaatg aaatgtacac agaatatctc
     2401 tccagtgcct tcgtgggtct gtccttccct actgtttgtg agctgtgttt tgtgaagctt
     2461 aagctcctga tgatagccat tgagtacaag tctgccaaca gagagagccg aagccgaaag
     2521 cgaatattaa ttaaccctgg gaaccacctt aagatccaag aaggtacttt aggatttttc
     2581 atcgcaagtg atgccaaaga agttaaaagg gcattttttt actgcaaggc ctgtcatgat
     2641 gacgtcacag atcccaaaag aattaaaaaa tgtggctgca ggcggcttga agatgagcag
     2701 ccgccaaccc tgtcaccaaa aaaaaaacaa cgtaatgggg gcatgaggaa ctcgcccaac
     2761 acctccccga agctgatgag gcatgacccc ttgttaattc ctggcaatga tcagattgac
     2821 aacatggact ccaatgtgaa aaagtacgac tccactggaa tgtttcactg gtgtgcaccc
     2881 aaggagattg agaaagtcat cttgactcga agtgaagctg ccatgactgt cctgagtggc
     2941 catgtcgtag tctgcatctt tggggatgtc agctcagccc tgattggcct ccggaacctg
     3001 gtgatgccac ttcgtgctag caactttcac tatcatgagc tcaaacacat tgtgtttgtg
     3061 ggctccattg agtacctcaa gagggagtgg gaaacactgc acaacttccc gaaagtgtcc
     3121 atattgcctg gtacaccatt aagtcgggct gatttaaggg ctgtcaacat caacctctgt
     3181 gacatgtgcg ttatcctgtc agccaatcag aataatattg atgatacttc gcttcaggac
     3241 aaggaatgca tcttggcgtc actcaacatc aaatctatgc agtttgatga cagcatcggg
     3301 gtcttgcagg ctaattccca aggattcaca cctcctggaa tggacagatc atcacccgac
     3361 aacagcccag tgcacgggat gttacgccag ccgtccatca caactggggt caacattccc
     3421 atcatcacgg aactcgctaa gccgggcaag ttgcctttgg tatcagtcaa tcaggaaaaa
     3481 aacagtggga cgcacattct aatgataacg gagttggtga atgataccaa tgttcagttt
     3541 ttggaccaag acgatgacga tgaccctgac acagagctgt acctcacaca gccctttgct
     3601 tgtgggacag catttgccgt cagcgtcctg gactcactca tgagcgcgac atacttcaat
     3661 gacaatatcc tcaccctaat acggaccctg gtgacaggag gagccacacc agagctcgag
     3721 gctctaatag ctgaggagaa tgcacttcga ggaggctaca gcactccgca gacattggcc
     3781 aacagggacc gttgccgagt ggcccagtta gccctgttag atggtccctt tgcagactta
     3841 ggggatggtg gttgttatgg tgatctgttc tgcaaagctc tgaaaacata taatatgctt
     3901 tgttttggaa tttaccggct gagagatgcc cacctcagca cccccagcca gtgtacaaaa
     3961 aggtacgtca tcaccaatcc tccctacgag tttgagctcg taccaacaga cctgatcttc
     4021 tgcctgatgc agtttgacca caacgctggc caatcccggg ccagtctgtc tcattcctcc
     4081 cactcctcac agtcgtccag taagaagagc tcctccgtcc actccatccc gtccacagca
     4141 aatcggccga accggcccaa gtccagggag tcccgcgaca aacagaacag aaaagaaatg
     4201 gtttacagat agatcgatag aactatagtg agtcgtatta cgtagatcca gacatgataa
     4261 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     4321 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     4381 acaacaacaa ttcggaggtt ctcatatggg cgggtcaacc ggtatcttac tcgaggcagc
     4441 tgtcgactct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     4501 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     4561 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     4621 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     4681 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     4741 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4801 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4861 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4921 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4981 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     5041 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     5101 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     5161 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     5221 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5281 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5341 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     5401 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     5461 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     5521 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     5581 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5641 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5701 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5761 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5821 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5881 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5941 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     6001 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     6061 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     6121 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     6181 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     6241 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     6301 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     6361 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     6421 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     6481 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     6541 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     6601 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     6661 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     6721 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6781 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6841 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6901 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6961 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     7021 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     7081 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7141 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7201 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7261 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     7321 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     7381 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7441 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     7501 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7561 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7621 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7681 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7741 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7801 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7861 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7921 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7981 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     8041 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     8101 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     8161 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     8221 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     8281 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     8341 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     8401 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     8461 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     8521 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     8581 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     8641 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     8701 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8761 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8821 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8881 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     8941 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     9001 gtagtgcgcg agc
//
