LOCUS       sequence-191155-        4896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To determine if structural domains are important for nicotinic 
            acetylcholine receptor (nAChr) channel function via dose-response 
            curves from voltage-clamped oocytes to estimate EC50's and Hill Co.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4896)
  AUTHORS   Labarca C, Nowak MW, Zhang H, Tang L, Deshpande P, Lester HA
  TITLE     Channel gating governed symmetrically by conserved leucine residues 
            in the M2 domain of nicotinic receptors.
  JOURNAL   Nature. 1995 Aug 10. 376(6540):514-6.
  PUBMED    7637783
REFERENCE   2  (bases 1 to 4896)
  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: "Nature. 
            1995 Aug 10. 376(6540):514-6."
COMMENT     Sequence Label: pBS-AMVpl-delta-WT-nAchR
FEATURES             Location/Qualifiers
     source          1..4896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(33..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(143..164)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     355..374
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2607..2623
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2607..2623
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2665..2681)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2666..2682)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(2711..2729)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2750..2766)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2750..2766)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2763..2785)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2774..2790
                     /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(2798..2804,2805..2822,2823..2828))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2843..2864
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2981..2998)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3152..3740)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3232..3251)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3911..4702,4703..4771))
                     /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     4534..4553
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4772..4876)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
       61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcgt
      661 ttttattttt aattttcttt caaatacttc caccatggca gggcctgtgc tcacactggg
      721 gctgctggct gcccttgttg tgtgtgccct cccaggcagt tggggcctga atgaggaaca
      781 aaggctgatc cagcacctat tcaatgagaa gggctacgac aaagacctgc ggccggtggc
      841 tcgaaaggag gacaaagtgg atgtcgccct aagtctcacc ctctccaacc tcatctccct
      901 gaaagaagtg gaggagaccc tcaccaccaa cgtgtggata gatcatgcct gggtagacag
      961 ccgtctacag tgggatgcca acgacttcgg gaacatcact gtcctgcgcc tgccccctga
     1021 catggtgtgg ctaccagaga ttgtactgga gaacaacaat gatggctcgt tccagatttc
     1081 ttatgcctgc aatgtgcttg tctatgactc gggctatgtg acctggctgc cgcccgccat
     1141 cttccgctcc tcctgcccca tctcagtcac ctacttccct ttcgactggc agaactgctc
     1201 cctcaaattc agttcactca agtatacagc caaggagatc acacttagcc tgaagcagga
     1261 ggaggaaaac aaccgctctt accccattga gtggatcatc attgaccctg aaggtttcac
     1321 agagaacggt gagtgggaga tagtgcatcg ggcagccaag ctcaatgtgg accccagtgt
     1381 cccgatggac agcaccaacc accaagatgt caccttctac cttatcatcc gccgcaagcc
     1441 cctcttctac atcatcaaca ttctggtacc ctgcgtgctc atctccttca tgatcaacct
     1501 agtcttctac ctgccaggcg actgtggaga gaagacctcc gtggccatct cagtgctcct
     1561 ggcccaatct gtcttcctgc tgcttatctc caagaggctg ccggccactt ccatggccat
     1621 ccccttggta ggcaagttcc tgcttttcgg catggtgctg gtcaccatgg tggtggtgat
     1681 ctgtgtcatc gtactgaaca tccacttccg gacgcccagc acccatgtgc tgtctgaggg
     1741 agtcaagaag ttcttcctgg agaccctgcc caagctcctg cacatgtccc gcccagcaga
     1801 ggaagaccca ggccccaggg ctctcatccg gagaagcagc tctctgggat acatctgcaa
     1861 agcagaggag tatttctcac ttaagtcccg cagtgacctc atgtttgaga agcaatcaga
     1921 gcggcatggg ctggcccggc gcctcaccac agcccgcagg cctccagcaa gctctgagca
     1981 ggtccaacag gagctcttca atgagatgaa gccagctgtg gatggggcaa acttcatcgt
     2041 caaccatatg agagaccaaa acagttacaa tgaggagaag gacaactgga accaggtggc
     2101 ccgcacagtg gaccgtctct gcctgtttgt ggtgacaccc gtcatggtgg tgggcacagc
     2161 ctggatcttc ctgcagggtg tctacaacca gcccccactc cagcctttcc ctggggaccc
     2221 cttctcctat agcgaacagg acaagcgctt catctaaagc aggcatgagt agacagccag
     2281 gcagagagca gtctgagaac ggcacgtagt tcctgccctt gagcttgtgc ctagcgggtg
     2341 ctgtgtggct gggcagaggc catggtcggg agagctccct tccggaaaca attcctttct
     2401 aaataaagga gggagacact tgagatgggc taagggtgga cgtggtggag agggaggctc
     2461 cagggttgtg aagagaagca ggggccttgc tcagggggcc agagaggtga ccttgggact
     2521 ggcttcctgg gaacagtcct gtgagagatc ccccaacctg agctgctcct ccgcctggct
     2581 gtcttccaga gcaggtcaga attcgctcta gaactagtgg atcccccggg ctgcaggaat
     2641 tcgatcagat ctgatcaagc ttatcgatac cgtcgacctc gagggggggc ccggtaccca
     2701 gcttttgttc cctttagtga gggttaattt cgagcttggc gtaatcatgg tcatagctgt
     2761 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     2821 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2881 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2941 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3001 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3061 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3121 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3181 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3241 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3301 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3361 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3421 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3481 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3541 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3601 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3661 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3721 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3781 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3841 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3901 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3961 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4021 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4081 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4141 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4201 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4261 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4321 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4381 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4441 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4501 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4561 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4621 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4681 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4741 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4801 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4861 aaataggggt tccgcgcaca tttccccgaa aagtgc
//
