LOCUS       sequence-41625-e        6716 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 6716)
  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 6716)
  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: mg177
FEATURES             Location/Qualifiers
     source          1..6716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3540..3556)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3540..3556)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     3'UTR           3579..3704
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(3784..3803)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3785..3803)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3813..3830)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3813..3829)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3822..3844)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3970..4425)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4107..4128)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4319..4338
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4452..4556
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4557..4625,4626..5417)
                     /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(4775..4794)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5588..6176
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6077..6096
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6330..6347
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6464..6485
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6500..6505,6506..6523,6524..6530)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6538..6554
                     /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     6543..6565
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6562..6578
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6562..6578
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6597..6617
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6599..6617
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     5'UTR           6658..6700
                     /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 gtctcctcac acccctctct ggcttaacga ctacttacgt
     1681 ggtgccggaa tggaaatgta cacggaaagt ctgagtcctt cgtttgcgaa tatgtcattt
     1741 ccagaagctg caaacctatt 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 caacagagtt cgacgagtga tactcatctc
     2161 aacacgaaat cccttcgatt tgcctatgaa atcaaaaaac tcatgccatc ttccggagga
     2221 cgccgaaatt cgatgagtat tccgccagac ggcaggggtg tcgattttag caaggatttt
     2281 gagcagcaat tccaagacat gaaatatgac agtactggaa tgttccactg gtgtccatcc
     2341 cgtaacctag aggactgtgt tctggagcgc caccaggccg ctatgaccgt actcaacggc
     2401 cacgtggtgg tttgtttgtt cgctgatcag gattctccgc taattgggct ccggaacttc
     2461 ataatgccac taagaagctc aaacttccat tatcatgagc tcaaacacgt ggttattgtg
     2521 ggagacctgg aatatttgag gaaagagtgg aagacgctgt acaacttgcc gaaaatctcg
     2581 attttgaatg gctccccgct atctcgagca gatttacgcg ccgtcaacat taacctctgc
     2641 gacatgtgtg tcataatctc tgcccgggtc ccaaataccg aagataccac tttggcagat
     2701 aaggaagcga ttctggcatc attgaacatt aaagctatgc aatttgatga tacccttggc
     2761 tttttcccga tgaggcatca aactggcgat cggagcccgc tgggctcccc gatttccatg
     2821 cagaagaagg gcgccaagtt tgggaccaat gtgccgatga ttacggagct tgttaacgac
     2881 tcaaatgtgc aattcctcga ccaggacgac gacgacgatc cggacaccga gctctacctc
     2941 acgcagccct tcgcctgcgg aactgccttc gccatttcag tgttggattc tttgatgagt
     3001 actacctact tcaacgactc ggccctgacc ctcatccgaa ctctggtcac tggtggagcc
     3061 acaccggagc ttgaacttat tctggcagaa ggcgccgggc tccgaggtgg ctacagtacc
     3121 ccggagaccc tctcaaacag agatcggtgc cgaattgctc agatttcgct gcaggataac
     3181 ccgtacgatg gagttgtgca caacactacc tatggtgcaa tgttcacgat agctctgagg
     3241 aggtacggcc aactttgtat cgggctgtat aggcttcacg accaggataa tccggactct
     3301 atgaagcgat atgttatcac taatcctcca gcagaattga ggattaaaaa tactgattac
     3361 gtctacgtac ttgaacaatt cgatccggga ctggagtacg agccgggcaa acgacacttt
     3421 tagggcctaa ttttatccga ttttcttgct acaaactacg ggttatccga ctccgaagga
     3481 ccaataatat ttgcaaatgc tcatttttag gacttttttt gaattcctgc agcccggggg
     3541 atccactagt tctagactcg aggatctggt taccactaaa ccagcctcaa gaacacccga
     3601 atggagtctc taagctacat aataccaact tacactttac aaaatgttgt cccccaaaat
     3661 gtagccattc gtatctgctc ctaataaaaa gaaagtttct tcacattcta aaaaaaaaaa
     3721 aaaaaaaaaa aaaaaccccc cccccccccc ccggggncgc caccgngggg gagctccaat
     3781 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
     3841 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     3901 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     3961 ggcgaatgga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     4021 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     4081 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     4141 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     4201 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     4261 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     4321 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     4381 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     4441 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4501 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4561 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4621 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4681 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4741 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4801 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4861 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4921 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4981 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5041 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5101 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5161 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5221 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5281 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5341 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5401 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     5461 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5521 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5581 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5641 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5701 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     5761 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5821 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5881 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5941 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6001 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6061 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6121 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6181 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6241 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     6301 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     6361 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     6421 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     6481 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     6541 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa
     6601 ttaaccctca ctaaagggaa caaaagctgg gtaccgggcc ccccctcgag gtagcttctt
     6661 gttctttttg cagaagctca gaataaacgc tcaactttgg cagaatcgat cccacc
//
