LOCUS       sequence-41622-e        6650 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6650)
  AUTHORS   Johnson BE, Glauser DA, Dan-Glauser ES, Halling DB, Aldrich RW, 
            Goodman MB
  TITLE     Alternatively spliced domains interact to regulate BK potassium 
            channel gating.
  JOURNAL   Proc Natl Acad Sci U S A. 2011 Nov 2.
  PUBMED    22049343
REFERENCE   2  (bases 1 to 6650)
  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"; journalName: "Proc Natl 
            Acad Sci U S A. 2011 Nov 2."
COMMENT     Sequence Label: mg174
FEATURES             Location/Qualifiers
     source          1..6650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3474..3490)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3474..3490)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     3'UTR           3513..3638
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(3718..3737)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3719..3737)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3747..3764)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3747..3763)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3756..3778)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3904..4359)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4041..4062)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4253..4272
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4386..4490
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4491..4559,4560..5351)
                     /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(4709..4728)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5522..6110
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6011..6030
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6264..6281
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6398..6419
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6434..6439,6440..6457,6458..6464)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6472..6488
                     /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)."
     primer_bind     6477..6499
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6496..6512
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6496..6512
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6531..6551
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6533..6551
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     5'UTR           6592..6634
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
ORIGIN
        1 atgggcgaga tttactcgcc ttcgcagtcg aagggcttca atcaaccata tggatacccg
       61 atgaattgta accttagtcg agtattcatg gagatgactg aagaagatcg aaagtgcctg
      121 gaggagcgca aatattggtg ttttttactg agcagcatca ccacattctg cgcctcaatg
      181 atattggtcg taatatggcg agtggtgaca cacctgtgct gtcaacgccg agaaaaagag
      241 tttgtggagc ccatacctgc tccggaagcc gtccaaataa atatgaacgg gtcgaagcac
      301 gcgccaagtg aaaccgaccc gtttttgaag caacaagagg aaaaacactt gggatggatg
      361 acagaagcca aggactgggc tggcgagctg atatccggac aaagtctgac gggacggttt
      421 ctggtacttt tggtgttcat tttatcgatt ggatcgttga ttatttattt ttacgacgct
      481 agtttccaaa acttccaagt agaaacgtgc atcccgtggc aggacagtcc ctcgcaacaa
      541 atcgacctgg gcttcaacat attttttctc gtctattttt tcattagatt catcgccgcg
      601 tccgacaaag tttggtttct gcttgaaatg tacagctgga ttgatttttt cacgattccg
      661 ccaagcttcg tggcaattta cttgcagagg aattggctcg gattccgttt cctccgtgct
      721 cttcgcctca tgaccgtacc cgacattcta cagtacctca acatcctgaa aacatcttca
      781 tcaatccgat tgacacagtt ggtcacaatt ttcgtggcgg tttgtctgac tggagccggc
      841 ttggtgcact tactggaaaa ttctggggac ttcttcaaag gatttataaa tccgcacaga
      901 atcacttatg cggactctgt gtactttgta ctggtcacaa tgtctactgt cggatatgga
      961 gatatctatt gtacgacgct ttgcggccga cttttcatga ttttcttcat tctgttcggt
     1021 ttggccatgt ttgcaagtta cgtaccagaa attgccgatt tgattggaaa ccggcaaaaa
     1081 tacggtgggg agtacaaagg agagcacggg aagaagcaca tagtggtctg tggccatatc
     1141 acctacgatt cggtgtccca ttttcttcaa gatttcctac acgaggaccg tgatgacgtg
     1201 gatgtcgaag tggtgttttt gcatcgtgtc gtgccggatt tggagctgga aggcttgttt
     1261 aagcggcatt tcacaaaagt cgaatttttc acggggactg tcatggattc tctggatctt
     1321 agcagagtca agatcgggga cgccgacgcc tgcctggtcc ttgccaacaa gtacagtacc
     1381 aatccggacg ccgaggacgc cgcaaacatt atgcgtgtaa tttcgatcaa aaactactca
     1441 tcggacatca gagttattgt gcaactaatg caatatcaca ataaggccta ccttctaaat
     1501 atcccctcct gggactggaa acgaggtgat gacgtcatct gtctcgcgga gcttaagctc
     1561 ggattcattg ctcaaagctg tctggcaccc ggattctcta cgatgatggc taatttattt
     1621 gcaatgagaa gtttcaaaac gtcacaaacc acaccggact ggctgaacct gtacctttgc
     1681 ggtgccggaa tggaaatgta caccgacacg ctatcccact cgttcgtcgg aatgactttt
     1741 cctgaagctg tcgacctatt attcaaccgc ctggggcttc ttcttctggc gattgagctt
     1801 aaagatgagg aaaataaaga atgcaatatt gcaatcaatc caggcccaca tattgtcatc
     1861 cagcctcaaa ctcaaggatt tttcattgct cagagcgctg acgaggtgaa gcgggcgttt
     1921 ttctggtgta agcagtgtca tgacgatatt aaggacgttt ccttgatcaa aaaatgtaaa
     1981 tgcaaaaatt tggccctctt ccgtcgcaac accaaacact cgacagcggc ccgtgcaaga
     2041 gccaccgatg tgctccagca gttccagcca caagcgccag caggtccgat gggccatttg
     2101 ggccaacaag ttcagctaag gatgattaat caacagaggc catcttccgg aggacgccga
     2161 aattcgatga gtattccgcc agacggcagg ggtgtcgatt ttagcaagga ttttgagcag
     2221 caattccaag acatgaaata tgacagtact ggaatgttcc actggtgtcc atcccgtaac
     2281 ctagaggact gtgttctgga gcgccaccag gccgctatga ccgtactcaa cggccacgtg
     2341 gtggtttgtt tgttcgctga tcaggattct ccgctaattg ggctccggaa cttcataatg
     2401 ccactaagaa gctcaaactt ccattatcat gagctcaaac acgtggttat tgtgggagac
     2461 ctggaatatt tgaggaaaga gtggaagacg ctgtacaact tgccgaaaat ctcgattttg
     2521 aatggctccc cgctatctcg agcagattta cgcgccgtca acattaacct ctgcgacatg
     2581 tgtgtcataa tctctgcccg ggtcccaaat accgaagata ccactttggc agataaggaa
     2641 gcgattctgg catcattgaa cattaaagct atgcaatttg atgataccct tggctttttc
     2701 ccgatgaggc atcaaactgg cgatcggagc ccgctgggct ccccgatttc catgcagaag
     2761 aagggcgcca agtttgggac caatgtgccg atgattacgg agcttgttaa cgactcaaat
     2821 gtgcaattcc tcgaccagga cgacgacgac gatccggaca ccgagctcta cctcacgcag
     2881 cccttcgcct gcggaactgc cttcgccatt tcagtgttgg attctttgat gagtactacc
     2941 tacttcaacg actcggccct gaccctcatc cgaactctgg tcactggtgg agccacaccg
     3001 gagcttgaac ttattctggc agaaggcgcc gggctccgag gtggctacag taccccggag
     3061 accctctcaa acagagatcg gtgccgaatt gctcagattt cgctgcagga taacccgtac
     3121 gatggagttg tgcacaacac tacctatggt gcaatgttca cgatagctct gaggaggtac
     3181 ggccaacttt gtatcgggct gtataggctt cacgaccagg ataatccgga ctctatgaag
     3241 cgatatgtta tcactaatcc tccagcagaa ttgaggatta aaaatactga ttacgtctac
     3301 gtacttgaac aattcgatcc gggactggag tacgagccgg gcaaacgaca cttttagggc
     3361 ctaattttat ccgattttct tgctacaaac tacgggttat ccgactccga aggaccaata
     3421 atatttgcaa atgctcattt ttaggacttt ttttgaattc ctgcagcccg ggggatccac
     3481 tagttctaga ctcgaggatc tggttaccac taaaccagcc tcaagaacac ccgaatggag
     3541 tctctaagct acataatacc aacttacact ttacaaaatg ttgtccccca aaatgtagcc
     3601 attcgtatct gctcctaata aaaagaaagt ttcttcacat tctaaaaaaa aaaaaaaaaa
     3661 aaaaaaaaac cccccccccc ccccccgggg ncgccaccgn gggggagctc caattcgccc
     3721 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     3781 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     3841 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     3901 tggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3961 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     4021 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     4081 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     4141 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     4201 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     4261 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     4321 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     4381 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4441 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4501 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4561 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4621 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4681 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4741 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4801 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4861 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4921 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4981 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5041 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5101 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5161 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5221 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5281 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5341 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5401 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5461 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5521 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5581 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5641 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     5701 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5761 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5821 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5881 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5941 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6001 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6061 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6121 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6181 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     6241 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     6301 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     6361 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     6421 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     6481 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
     6541 ctcactaaag ggaacaaaag ctgggtaccg ggccccccct cgaggtagct tcttgttctt
     6601 tttgcagaag ctcagaataa acgctcaact ttggcagaat cgatcccacc
//
