LOCUS       sequence-183032-        4629 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 4629)
  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 4629)
  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-gamma-WT-nAchR
FEATURES             Location/Qualifiers
     source          1..4629
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    17..38
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(53..58,59..76,77..83)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    91..107
                     /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     96..118
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     115..131
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     115..131
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        152..170
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1969..1988)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1970..1988)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1995..2012)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1995..2011)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2004..2026)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2153..2608
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2240..2259)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2450..2471
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2634..2738
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2739..2807,2808..3599)
                     /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(2957..2976)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3770..4358
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4259..4278
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4512..4529
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
       61 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
      121 acagctatga ccatgattac gccaagctcg aaattaaccc tcactaaagg gaacaaaagc
      181 tggtaccgcg gccgcttttt tttttttttt ttttcccctc tgtctcttta tttctctatc
      241 ttctcccagg aagccttggg tatttcccca agagcacact cagcatcaac acgggataca
      301 gctctacaaa cagcatggcc ctcatagctg actccagggc ccaggaggga ttggttggct
      361 tagtctggca aaggcaggta ggggcggggg tctccgggga acggcaggtc aggcacttgg
      421 ttgtagtggg ccatgaggaa gatgccagca gtgccacaga tgaagaggga gagcatggct
      481 aggaagcaga ctcggtccag cactcggccc accagcaacc actcctcgtt cccactgtca
      541 aagtgactct gctggcgtcg ggcacgagcc atgaggttac aggcatccac acaggcctgg
      601 atggctgggg aggcttgctt caggctgcca cagaactcct ggctctgcct cacttctgga
      661 ccattctcta gcttctccaa tactgcctgc actaatccat tgcgctgcct ttgcctaaag
      721 aggagttcgc ttcgaggcag acagaggtcc ccttcctctc gagccatgat gggccaccct
      781 gaggaagagc cattctggag tcggaaccgg gcatcctgga cagcagctgg agctagtggg
      841 cgcacatgca tccgtaacag ctggggcagg agcctcagga acaacttgcg gaccccacgg
      901 gccatggagt gtgtgtgggg ggaccgcaag gacacattga gcacgaccac agagttcacg
      961 acgatgagga tggtcaccac catgaggaag gtcaggtact tgctgatgag tggcactgcc
     1021 tgggaggtct caggcacctt cttagccaca aggaaaagga agacagtctg ggccaggagc
     1081 acgttggtgg ccactgtgca tttctggccg cccgccttag caggaaggaa gtagatgagg
     1141 atggcgactg aggagatgag cacacagggg gcgatgatgt tgatgacgta gaagaggggc
     1201 ttgcgctgga taagcaggta gaacaccacc ttctggtggc ccgcctcctc tgcaggagcc
     1261 acggagtcca ggagcatttt agccggtcgg tgccggatgg cccactcccc attctctgtg
     1321 aaagcctccg ggtcaatgaa gatccactca atggcttgcc catcctcctg gctcagctgc
     1381 aagttgatct cactggtgct gtaagtctgg gattggaaga tgagggaaca gttctgccaa
     1441 tcgaagggga agtaggtgac agagatggag caggaggagc ggaagatggc aggcggcagc
     1501 cagtagatac aaccgtcggg ggacacgagg acattgcagt agagagccac ctcgaagaca
     1561 ccgtccacat tgttctccag gacgatatct ggccgccaga ccatagtaga tggcaccctc
     1621 aatatccaca ggccttcgta gtcttttggg tcccagcgca ggcgatagtc gcaccattgc
     1681 atctctatcc agacgttagt tgtgagggcc tcctctcgtt cattcaggga gatgaggttg
     1741 gtcaaggtaa gcttcaggct gacattgacc acatctgagt cgcgctcagc cggccgcagg
     1801 tgggggtcgt agtttcgcat caggtccgca agcagacgct cctcttggtt gcggctctgg
     1861 gcccccaggc agacagccaa cagcagcaag aggagatgag gtctctggcc cccttgcatg
     1921 gtggaagtat ttgaaagaaa attaaaaata aaaacgagct ccaattcgcc ctatagtgag
     1981 tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     2041 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     2101 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     2161 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2221 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2281 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2341 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2401 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2461 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2521 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2581 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     2641 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2701 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2761 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2821 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     2881 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     2941 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3001 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3061 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3121 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3181 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3241 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3301 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3361 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3421 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3481 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3541 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3601 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     3661 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     3721 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3781 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3841 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3901 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3961 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4021 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4081 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4141 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4201 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4261 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4321 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4381 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4441 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     4501 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     4561 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     4621 aagcgggca
//
