LOCUS       sequence-128800-        6644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For in vitro transcription of alpha-bungarotoxin mRNA that can be 
            injected into embryos..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6644)
  AUTHORS   Swinburne IA, Mosaliganti KR, Green AA, Megason SG
  TITLE     Improved Long-Term Imaging of Embryos with Genetically Encoded 
            alpha-Bungarotoxin.
  JOURNAL   PLoS One. 2015 Aug 5;10(8):e0134005. doi: 
            10.1371/journal.pone.0134005. eCollection 2015.
  PUBMED    26244658
REFERENCE   2  (bases 1 to 6644)
  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: "PLoS One.";
            date: "2015-08-5"; volume: "10(8):e0134005. doi"; pages: " 
            10.1371/journal.pone.0134005. eCollection 2015"
COMMENT     Sequence Label: pmtb-t7-alpha-bungarotoxin
FEATURES             Location/Qualifiers
     source          1..6644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      353..941
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     842..861
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1095..1112
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1229..1250
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1265..1270,1271..1288,1289..1295)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1303..1319
                     /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     1308..1330
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1327..1343
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1327..1343
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1362..1382
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1364..1382
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     1419..1435
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1419..1435
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    join(1469..1480,1481..1668)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     promoter        4384..4402
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4384..4401
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     4410..4429
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4410..4428
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4788..4922
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4812..4831)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4866..4885
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(5014..5151,5152..5163)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(5222..5239)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5222..5238)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5231..5253)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5380..5835
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5467..5486)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5677..5698
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5861..5965
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5966..6034,6035..6644,1..182)
                     /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(6184..6203)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
       61 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
      121 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
      181 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
      241 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
      301 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
      361 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
      421 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
      481 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
      541 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
      601 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
      661 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
      721 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
      781 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
      841 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
      901 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
      961 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     1021 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     1081 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     1141 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     1201 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     1261 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     1321 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
     1381 gggaacaaaa gctggagctc caccgcggtg gcggccgctc tagaactagt ggatctgctg
     1441 ggcttgctga aggtaggggg tcaagaacca gaggtgtaaa gtacttgagt aattttactt
     1501 gattactgta cttaagtatt atttttgggg atttttactt tacttgagta caattaaaaa
     1561 tcaatacttt tacttttact taattacatt tttttagaaa aaaaagtact ttttactcct
     1621 tacaatttta tttacagtca aaaagtactt attttttgga gatcacttgg gcccggctcg
     1681 agggcctagt cattacctat actcaacgtt tactctgaga aacttccata ttttagaggt
     1741 aaaatgctca gaattgtaaa aagtcttgtc aaaatacttg gaatttgcca aaaacaaaat
     1801 aaaagtcaca tcacctcatt gtccttggta tcagcaagca cttttaacac aatcagctgt
     1861 ttttgtttat aactcccact gaaggccatg tgcattactc ccaagttaca cagtaaaaag
     1921 tcacatgtag tttatcatga tcactgctgc tcaagcacag ctttatctgt tattcagtgt
     1981 ggcaatatct gaaatgtgtg caaaatatta tttattgtag ctgatcacca acactttcag
     2041 tgtttttttt tttctacttt taatgagaat tatgttctta ggatttatcc ttataaacct
     2101 ctttaaaaag aaatgttatg tctgcataga cacgacatga aattatgaac agaaaattta
     2161 actatttaag atcaagactt gacatataac atttctaatg aacattaaac ctgaggtagg
     2221 cccacttgca aaaatgtaac tgctatcact cactttatac atcattgtgt aacttgtaca
     2281 cacaaaaata gtgaagcaat gtggcatgga atgcaggtcc cattcattca aagttattaa
     2341 agcactttct taaagtgcat tactactgag ctaagaccag ctagttgctc atacaacata
     2401 ctacaagtag tgtcatccta atttggataa aaaaaatgtc cattttatac atggaaaagt
     2461 actcattaga tcccccattg tttgtattac ggtatttcgt gaacacaaga ggtaaatgac
     2521 ctacagagct gctgttgtgt tagattgaaa acacaacaca ggatcatgga ggcatccatc
     2581 attatgacgc attctgccac ttgtaactct gcacagactt tttgaaaagt ttaattgagt
     2641 cttggcgttg tcgcagaaaa atacgaaaaa ttctgtgtta aaatttgcaa ataaaaattc
     2701 cagttttata ggaaatcgcc atggccttgt tctttaacca gttaagcttc cccttctttc
     2761 actctcaagt tgcaagaagc aagtgtagca atgtgcacgc gacagccggg tgtgtgacgc
     2821 tggaccaatc agagcacaga gctccgaaag tttacctttt atggctagag ccgggtatgt
     2881 gccgtcatat aaaagagcgc gcccagcttt tcagcctcac tttgagctcc tccacacgca
     2941 gctagtgcgg aatatcatct gcttgtaacc cattctctta agtcgacaac cccccaaacc
     3001 caaggtgagt tggttttaca gtttttaagc gtttaattat agtttctatc tattaatgtt
     3061 aagtaaaatg tgtaaatgga taaatatgat ttgatgaagt ctttagacag ttcataacgg
     3121 tctctttgta gttatctact aggtttgatt tataagtggt ggttcttttg ataatctatg
     3181 ttacatttaa taaatattgg ctgtatctta acttttttgt agcgttagtg gatgtggcag
     3241 gtgagaatgt cggtaattta acgtgaccag tcgtaggcac gacattgaac attgaatagg
     3301 ccggtgtgaa atgggtgtta agtcttaata taaagttgtg ttctggctaa tctctcttta
     3361 acagccataa atgtgatttt taactacagt tttgagggtg ctggtgagca tgttgcacac
     3421 ttgatggata gccggcatgg gaagttcttt gtgcaggcag tgctgcagca gggtgtgacc
     3481 tactttagct agccggctaa ccagctctca tctgatgtaa cgtaaacccc atcaatctaa
     3541 catcggctca ttcgtgcttt ataagatcga agatcttgtt attagtaacg ggtattggca
     3601 tttagtttta acttggttaa ttttcttttt gttatgtgtt ctatttaaat gtagcctgca
     3661 gccccctctg tctttaatga agctgaagat taacgttaac gttagtgaag tgttgtatcc
     3721 gctatgaaag atttgaattt tagaggtgtg gtccacttta ataacggctg cggagttctc
     3781 tggctgcact tcagccgctg tgactgtcga agcagagccc atcagttttt caggatctcg
     3841 gtgatttttg aggctagact atgaactgaa ccgactgaga gtgaccgtcc agaaatgctt
     3901 tagtgtgaag tctccgctca cgaggagtcc gtggactgac tgcagatctg tgacgcagta
     3961 acacccggcg gcagacaccc gttgaaatgc ggttgtgtaa ttgatcgcag gcgagggtgg
     4021 aagaggatgt gaaacttcat ttgtgtaaaa tttagggagt ggtcctggtg tgatgaatgt
     4081 cgaaatctgt tcctttttac tgagccctac gactctggct gagtgccaca ccgccggcag
     4141 ccgcaaagcg gctcaaacca ttgcctttta tggtaataat gagagaatgc agagggactt
     4201 cctttgtctg gcatatctga ggcgcgcgct gtcactagcg cccaccagcg gtcagactgt
     4261 agaatgcaga gcaaaacagg aagttgactc cagatggtca catgcttgct gaaacgcctc
     4321 tccctggcag cagtgcaatt ctaacagttt ccctttcttt tacagttcag ccatcctagg
     4381 cctatttagg tgacactata gaagagtact aatacgactc actataggga gaagctactt
     4441 gttctttttg caggatccgc caccatgaag accctgctgc tgaccctggt ggtggtgaca
     4501 atcgtgtgcc tggacctggg atacaccatc gtgtgccaca ccaccgccac cagccctatc
     4561 agcgctgtga cctgccctcc tggcgagaac ctgtgctaca gaaagatgtg gtgcgacgcc
     4621 ttctgtagca gcagaggcaa ggtggtcgag ctgggatgcg ccgctacctg ccctagcaag
     4681 aaaccttacg aagaggtgac ctgctgtagc accgacaagt gcaaccctca ccctaagcag
     4741 cgccctggat gatctgcagt aagaattcat acgtatccgg aaccggtgat ccagacatga
     4801 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     4861 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     4921 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     4981 tttgatatcc cgggtttaaa cgcatgcaga tctaatactc aagtacaatt ttaatggagt
     5041 acttttttac ttttactcaa gtaagattct agccagatac ttttactttt aattgagtaa
     5101 aattttccct aagtacttgt actttcactt gagtaaaatt tttgagtact ttttacacct
     5161 ctgtcaagaa ccatatgccg gtacccaatt cgccctatag gacgtcgtat tacgcgcgct
     5221 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     5281 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     5341 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     5401 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     5461 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     5521 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5581 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5641 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5701 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     5761 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5821 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5881 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5941 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     6001 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     6061 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     6121 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     6181 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     6241 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6301 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6361 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6421 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6481 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     6541 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6601 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggct
//
