LOCUS       sequence-189664-        5063 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 5063)
  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 5063)
  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-beta-1WT-nAchR
FEATURES             Location/Qualifiers
     source          1..5063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(11..29)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(50..66)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(50..66)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(63..85)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    74..90
                     /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(98..104,105..122,123..128))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    143..164
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(281..298)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(452..1040)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(532..551)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1211..2002,2003..2071))
                     /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     1834..1853
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2072..2176)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2202..2657)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2339..2360)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2551..2570
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2784..2806
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2798..2815
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2799..2815
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2822..2841
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2822..2840
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4979..4995
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4979..4995
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5037..5053)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5038..5054)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 gcttttgttc cctttagtga gggttaattt cgagcttggc gtaatcatgg tcatagctgt
       61 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
      121 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
      181 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
      241 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
      301 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
      361 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
      421 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
      481 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
      541 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
      601 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
      661 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
      721 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
      781 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
      841 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
      901 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
      961 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1021 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     1081 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     1141 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     1201 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     1261 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     1321 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     1381 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     1441 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     1501 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     1561 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     1621 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     1681 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     1741 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     1801 gaccgagttg ctcttgccca gcgtcaatac gggataatac cgcgccacat agcagaactt
     1861 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     1921 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     1981 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     2041 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     2101 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     2161 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     2221 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2281 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2341 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2401 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2461 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gggcttgacg
     2521 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2581 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2641 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     2701 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     2761 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     2821 gtaatacgac tcactatagg gcgaattgga gctcgttttt atttttaatt ttctttcaaa
     2881 tacttccacc atggccttag gggcgctgct tctgcttcta ggggtactcg ggacgcccct
     2941 cgccccaggc gcccgcggga gcgaagccga aggccaactg attaagaaac ttttctccaa
     3001 ctatgatagc tcggtgaggc cggcgaggga ggtgggagac cgcgtcgggg tcagcatcgg
     3061 cctcaccctg gcgcaactca tcagcctgaa cgagaaggat gaagaaatga gcacaaaggt
     3121 gtacttagac ctggagtgga ccgactacag gttaagctgg gaccccgcag aacatgatgg
     3181 catcgattct ctccgcatca ctgctgaatc cgtttggctc cctgatgtgg tgctgctgaa
     3241 caacaatgac ggaaatttcg acgttgccct ggacatcaat gttgtggtat ctttcgaggg
     3301 ctctgtgcgc tggcaacccc ctggcctgta ccgcagcagc tgcagcatcc aggtcaccta
     3361 cttccccttt gactggcaga actgcaccat ggtgtttagt tcctacagct atgacagctc
     3421 tgaggtcagc ctgaagacgg gcctggatcc tgaaggagag gagaggcagg aagtctacat
     3481 tcatgaaggg accttcattg agaatggcca atgggagatt atccacaaac cttctaggtt
     3541 aatccaactt ccaggggatc agagaggtgg gaaggaaggg catcatgagg aagttatctt
     3601 ctatctcatt attcggcgga agcctctctt ctacctggtc aacgtcattg ccccatgcat
     3661 cctcatcacg ctcctggcca tctttgtctt ctacctccca ccagatgcag gagagaagat
     3721 ggggctctcc atctttgccc tgctgacgct cactgtgttc ttgctgctgt tggccgacaa
     3781 agtgcctgag acctccctgg cggtgcccat cattatcaag tacctcatgt ttaccatggt
     3841 ccttgtcacc ttctccgtta tccttagtgt tgtggtcctc aacctgcatc atcgctcacc
     3901 ccacacccac caaatgccct tttgggtccg ccagatcttc attcacaagc tccctccata
     3961 cctgggtctg aagaggccca aacccgagag agaccaactc cctgaaccac atcactctct
     4021 ttctccaaga agtggctggg gcagaggaac tgatgaatat ttcatccgga agcctccaag
     4081 tgattttctt ttccctaaac ttaacaggtt tcagcctgaa tcatctgccc cggacctgcg
     4141 acgatttatc gatggtccaa ccagggctgt aggtctgcct caggagctac gagaggtcat
     4201 ttcctcaatc agctacatgg cccgacagct tcaggaacag gaggaccacg acgcactgaa
     4261 ggaggactgg cagtttgtgg ccatggttgt ggaccgtctt tttctgtgga cattcatcgt
     4321 tttcacaagc gtcgggaccc tagtcatttt cctagatgcc acgtaccatt tgccccctcc
     4381 tgaacctttc ccttaaggac cctccggtca tttttgccaa atgctgcagg tactctcaag
     4441 ctcaatagcg cttggcgctt gtaagaatct tgtttctggg aagacgttca agtgagactt
     4501 ggaggctgag catggtgcct tggacccacc cgaaatgtcc ttctacttat tatttagagt
     4561 tttgtttttg ttgtttttga gacagagtct cactctgtgg cccaggctag cctggaactg
     4621 tggcgtccag gctggtctca aactcccaac aatgttactt ccttagccta gcaattgttg
     4681 gaatcacaga cctggtaaac cagcacactt ggagtttgtt tcttgaaact acatagaatg
     4741 tcacagctta gatctctagt gaaatagaat tataaccaag tagtagactg taagcctttt
     4801 gaggacagga gatctgttgg ctactgaatt atccccagca atgtgtacag gtcctagccc
     4861 ataagtgctt aacatttgtt gaagagaggg aaaataaatc tggaaggaaa tataaatatc
     4921 gcctacaatt gcacgtatta aaattaaaat gtagaaaatg attaaaaccc gaattcgctc
     4981 tagaactagt ggatcccccg ggctgcagga attcgatcag atctgatcaa gcttatcgat
     5041 accgtcgacc tcgagggggg gcc
//
