LOCUS       sequence-218939-        5863 bp    DNA     circular SYN 14-APR-2025
DEFINITION  mouse BK beta1 subunit in mammalian expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5863)
  AUTHORS   Wang B, Brenner R
  TITLE     An S6 mutation in BK channels reveals beta1 subunit effects on 
            intrinsic and voltage-dependent gating.
  JOURNAL   J Gen Physiol. 2006 Dec;128(6):731-44. doi: 10.1085/jgp.200609596.
  PUBMED    17130522
REFERENCE   2  (bases 1 to 5863)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1085/jgp"; 
            journalName: "J Gen Physiol"; date: "2006-12"; volume: "128"; issue:
            "6"; pages: "731-44"
COMMENT     Sequence Label: BK beta1
FEATURES             Location/Qualifiers
     source          1..5863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(583..1157,1158..1160,1161..1169)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(750..767)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     977..996
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1170..1172,1173..1175,1176..1889)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1170..1172,1173..1175,1176..1886)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1215..1236)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1476..1495)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1823..1844
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2012..2133
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2049..2068)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2103..2122
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2140..2595)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2277..2298)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2489..2508
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2622..2726
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2728..3085
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2728..2919
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2751..2771)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2936..3071
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3120..3914
                     /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             3120..3914
                     /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(3174..3193)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3784..3803
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4102..4121)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4146..4193
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4522..5110
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5011..5030
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5285..5588
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5589..5792
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5743..5763
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5858..5863,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggggaaga agctggtgat ggcccagaag cgcggagaga cacgagccct ctgcctggga
       61 gtggcaatgg tagtgtgtgc tgccatcacc tactacgtcc tgggtacaac tgtgctgccc
      121 ctctaccaga aaagtgtgtg gacccaggaa tccatatgtc acttgattga aactaatatc
      181 aaggaccagg aagagctgga gggcaagaag gtgccccagt acccatgcct ttgggtcaat
      241 gtatcagctg tgggcagatg ggccatgctg tatcacacgg aagacactcg ggatcaaaac
      301 caacagtgct cctatatccc caggaacctg gacaactacc agacagcctt ggcagatgtg
      361 aagaaggtca gagccaattt ctataagcac catgaattct attgcctttc tgcacctcaa
      421 gtcaacgaga ccagcgtcgt gtaccagcgc ctctacgggc cccaagtcct cctcttctcc
      481 ttcttctggc ccaccttcct gctgaccggg ggtcttattc ttattgccat ggtgaagctc
      541 aacaggtccc tatccatctt ggcggctcag aagtaggatc cgcccctctc cctccccccc
      601 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
      661 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
      721 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
      781 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
      841 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
      901 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
      961 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     1021 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct ttacatgtgt
     1081 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     1141 aaaaacacga tgataatatg gccacaacca tggtgagcaa gggcgaggag ctgttcaccg
     1201 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     1261 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     1321 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     1381 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1441 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1501 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1561 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1621 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     1681 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1741 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     1801 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1861 ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca taatcagcca
     1921 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     1981 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     2041 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2101 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     2161 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     2221 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     2281 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     2341 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     2401 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     2461 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     2521 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     2581 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2641 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2701 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     2761 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2821 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2881 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     2941 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3001 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3061 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     3121 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3181 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3241 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3301 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3361 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3421 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3481 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3541 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3601 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     3661 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     3721 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     3781 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     3841 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     3901 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     3961 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4021 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4081 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     4141 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     4201 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     4261 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     4321 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     4381 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4441 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4501 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4561 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4621 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4681 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4741 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4801 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4861 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4921 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4981 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5041 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5101 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5161 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     5221 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     5281 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     5341 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     5401 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     5461 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     5521 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     5581 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     5641 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     5701 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     5761 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc
     5821 gctaccggac tcagatctcg acggtatcga taagcttgcc gcc
//
