LOCUS       sequence-237102-        4499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express membrane-tethered alpha-bungarotoxin in Danio rerio.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4499)
  AUTHORS   Wan Y, Wei Z, Looger LL, Koyama M, Druckmann S, Keller PJ
  TITLE     Single-Cell Reconstruction of Emerging Population Activity in an 
            Entire Developing Circuit.
  JOURNAL   Cell. 2019 Oct 3;179(2):355-372.e23. doi: 
            10.1016/j.cell.2019.08.039. Epub 2019 Sep 26.
  PUBMED    31564455
REFERENCE   2  (bases 1 to 4499)
  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"; doi: 
            "10.1016/j.cell.2019.08.039"; journalName: "Cell"; date: "2019-10-3-
            3"; volume: "179"; issue: "2"; pages: "355-372.e23"
COMMENT     Sequence Label: pCS2.aBTX
FEATURES             Location/Qualifiers
     source          1..4499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        39..1016
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1051..1069
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1051..1068
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1170..1193
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1567..1701
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1591..1610)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1645..1664
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1814..1834)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1814..1832)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1853..1869)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1853..1869)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1866..1888)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1877..1893
                     /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(1901..1907,1908..1925,1926..1931))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1946..1967
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2084..2101)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2255..2843)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2335..2354)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3014..3805,3806..3874))
                     /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     3637..3656
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3875..3979)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4005..4460)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4142..4163)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4354..4373
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gatcggtgcg ggcctcttcg ctattacgcc agtcgaccgc caattcaata tggcgtatat
       61 ggactcatgc caattcaata tggtggatct ggacctgtgc caattcaata tggcgtatat
      121 ggactcgtgc caattcaata tggtggatct ggaccccagc caattcaata tggcggactt
      181 ggcaccatgc caattcaata tggcggacct ggcactgtgc caactgggga ggggtctact
      241 tggcacggtg ccaagtttga ggaggggtct tggccctgtg ccaagtccgc catattgaat
      301 tggcatggtg ccaataatgg cggccatatt ggctatatgc caggatcaat atataggcaa
      361 tatccaatat ggccctatgc caatatggct attggccagg ttcaatacta tgtattggcc
      421 ctatgccata tagtattcca tatatgggtt ttcctattga cgtagatagc ccctcccaat
      481 gggcggtccc atataccata tatggggctt cctaataccg cccatagcca ctcccccatt
      541 gacgtcaatg gtctctatat atggtctttc ctattgacgt catatgggcg gtcctattga
      601 cgtatatggc gcctccccca ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc
      661 ccattgacgt caataggacc acccaccatt gacgtcaatg ggatggctca ttgcccattc
      721 atatccgttc tcacgccccc tattgacgtc aatgacggta aatggcccac ttggcagtac
      781 atcaatatct attaatagta acttggcaag tacattacta ttggaagtac gccagggtac
      841 attggcagta ctcccattga cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac
      901 atccccattg acgtcaatgg ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa
      961 tgggcggtag gcgtgcctaa tgggaggtct atataagcaa tgctcgttta gggaaccgcc
     1021 attctgcctg gggacgtcgg agcaagcttg atttaggtga cactatagaa tacaagctac
     1081 ttgttctttt tgcaggatcc catcgattcg aattcgccac catgaaagca tttatactgt
     1141 tagctctatt cgctgtggca tacgctgctg actataagga cgatgatgat aagcttattg
     1201 tctgtcacac aacagctacc agcccaattt ccgcagtcac ttgcccaccc ggagagaacc
     1261 tctgctatag gaagatgtgg tgtgacgctt tctgtagtag cagaggcaag gtcgtagagt
     1321 tggggtgcgc tgccacttgt ccttcaaaaa agccttatga ggaggtgacc tgctgcagta
     1381 ccgacaagtg taaccctcat ccaaagcagc gaccaggagc tgctgccgga ggcgccttgt
     1441 gtaacggtgc tagcgccgtg caactcagta caaccgccgc tttctctact gctttattgg
     1501 caagtatatg gtcaagctac atgttgtgac tcgagcctct agaactatag tgagtcgtat
     1561 tacgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1621 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1681 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     1741 ggggaggtgt gggaggtttt ttaattcgcg gccgcggcgc caatgcattg ggcccggtac
     1801 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     1861 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccgggagca
     1921 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     1981 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     2041 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     2101 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2161 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2221 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2281 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2341 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2401 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2461 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2521 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2581 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2641 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2701 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2761 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2821 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2881 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     2941 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3001 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3061 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3121 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3181 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3241 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3301 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3361 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3421 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3481 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3541 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3601 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     3661 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     3721 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     3781 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     3841 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     3901 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     3961 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
     4021 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4081 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     4141 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     4201 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     4261 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     4321 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     4381 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     4441 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggc
//
