LOCUS       sequence-386860-        5785 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  zebrafish expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5785)
  TITLE     Koster Lab Plasmids 2022
REFERENCE   2  (bases 1 to 5785)
  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"
COMMENT     Sequence Label: pTol2-nbt300 2n-KalNFB-syt2a
FEATURES             Location/Qualifiers
     source          1..5785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(30..41,42..179)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     187..206
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        187..205
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             617..1057
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             1505..1876
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     polyA_signal    3195..3276
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3232..3251)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3286..3305
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(3382..3400)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    join(3407..3594,3595..3606)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3636..3652)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3636..3652)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3649..3671)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3660..3676
                     /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(3684..3690,3691..3708,3709..3714))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3729..3750
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3867..3884)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4038..4626)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4118..4137)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4797..5588,5589..5657))
                     /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     5420..5439
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5658..5762)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaatc ccatatgggc agaggtgtaa aaagtactca aaaattttac
       61 tcaagtgaaa gtacaagtac ttagggaaaa ttttactcaa ttaaaagtaa aagtatctgg
      121 ctagaatctt acttgagtaa aagtaaaaaa gtactccatt aaaattgtac ttgagtatta
      181 gatctgtaat acgactcact atagggacat agaatgcagg tacacgagca gcatccattt
      241 ccattccctc ccttgtcatt tgattggttc ccatccctac aatgctcagg gccagtggct
      301 ccacccacca ttacatcatc tgagcccagg accacagcaa gtgaatagga gagagcagca
      361 ccctccccaa ggcgtcacaa gtgtataaaa ggcagcagaa ggagcctctg acagcccaga
      421 ttcttagccg gctggaaacc atcggggtgt tgcacaaagg gcaagtaaag caacaagaaa
      481 gggccacacc atccactgat tacaccactg taagactcaa cccaatcaag cttggatccc
      541 ttcagcacca aggacagggc ctctaggaca ctagaggttt cagcaccaag gacagggcct
      601 cgaattcgcc gccaccatga aactgctctc atccatcgag caagcctgcg acatttgtcg
      661 gcttaagaag ctgaaatgct ccaaggaaaa gccgaaatgt gccaaatgcc tgaagaacaa
      721 ttgggaatgt cgttactctc ccaaaaccaa gcgaagtcca ctcacaaggg ctcatctgac
      781 cgaagtggag agcaggctag agagactgga acaactcttt ttgctcatct tccctagaga
      841 ggaccttgac atgatcctca agatggattc tctccaggat attaaagccc ttttgactgg
      901 cttattcgtc caggacaatg tgaacaaaga cgctgtgaca gaccgattgg caagtgtcga
      961 gaccgatatg cctctgacac tgagacagca cagaatcagc gctacttcct caagcgaaga
     1021 gtcttctaac aagggacaga gacagctgac tgtttcgagc aggtcgaggc ccatggagtt
     1081 ccagtacctg ccagatacag acgatcgtca ccggattgag gagaaacgta aaaggacata
     1141 tgagaccttc aagagcatca tgaagaagag tcctttcagc ggacccaccg acccccggcc
     1201 tccacctcga cgcattgctg tgccttcccg cagctcagct tctgtcccca agccagcacc
     1261 ccagccctat ccctttacgt catccctgag caccatcaac tatgatgagt ttcccaccat
     1321 ggtgtttcct tctgggcaga tcagccaggc ctcggccttg gccccggccc ctccccaagt
     1381 cctgccccag gctccagccc ctgcccctgc tccagccatg gtatcagctc tggcccaggc
     1441 cccagcccct gtcccagtcc tagccccagg ccctcctcag gctgtggccc cacctgcccc
     1501 caagcccacc caggctgggg aaggaacgct gtcagaggcc ctgctgcagc tgcagtttga
     1561 tgatgaagac ctgggggcct tgcttggcaa cagcacagac ccagctgtgt tcacagacct
     1621 ggcatccgtc gacaactccg agtttcagca gctgctgaac cagggcatac ctgtggcccc
     1681 ccacacaact gagcccatgc tgatggagta ccctgaggct ataactcgcc tagtgacagg
     1741 ggcccagagg ccccccgacc cagctcctgc tccactgggg gccccggggc tccccaatgg
     1801 cctcctttca ggagatgaag acttctcctc cattgcggac atggacttct cagccctgct
     1861 gagtcagatc agctcctaat ctagttctag acgcgccaac ccgctcctcc tccctccctc
     1921 cctccctccc tcctctcctt tattcacttt tcaacatgca caacacgtca tcaaatcccc
     1981 caatcagcac tgaattacag ccacgggcca ccagactgaa ggtgttaaag aagacgattc
     2041 ggtctcatca gcggctaaaa cgagatgagg gtgaaccgag agagactgtt tccacagatg
     2101 ttcttgaagg agggccgggc tcgtaggtgc ggttattaca catataagct cccattcaca
     2161 ccttaaaaca tccaacagca ttccaaagtt ttccatcggg gaccagggat agaaaacact
     2221 gcctcttcca ctgcctaatt taacacattc cttctagatt atctctggtc atctagatga
     2281 aatcagttct gttctgtttg ccgtgtctga aaagacagta aagagtaatt gaaaaaaaaa
     2341 atgaatatag aaaaatgggg actggagaga tatttggact agacgctcct gatccagcca
     2401 gaaggaacct ttcaggtgtc gattacactg cctgcgaaaa caacctcttc ttccattctt
     2461 cagtttccag ctgccggctg gcttcttcca ctgcccaaca tcctaacaaa cccctgacaa
     2521 aaaaggagac aagccttgaa acatcctgaa ataagcaatt cttagctgcc agggacgtct
     2581 tcctgttagc atcagtgtat ttgtttcttt tgctttgatt tgttttcttt ttttatttta
     2641 ttttcatggt aaatgtattc tgcctgagca ctttaaacga aggtgttttt gttccagtcg
     2701 atgcttcgtt ttgaacctgc ttttatattt attaactttt gttgtgtatt tttaatgttt
     2761 aatctttaaa ggaatagttc acctacacac acacacacac acacacacac acacacaaat
     2821 gaaatgaagt catcatttac tcacttttct ttcttttagc tgtggagcac aaaacgagat
     2881 gttttgaaga atgttcacgc tgttgttttt catgcaattt aagtaaacga agatcaaggg
     2941 atgttgagct atacaaatat tcattatatg ggcagcacgg tcgctggttc gagtcccggc
     3001 tgggccagtt ggcatttctg agtttgcata ttctccctgt gttggcatgg gtttcctcct
     3061 ggtgctccgt ttctcccaca gtccactaga tcataatcag ccataccaca tttgtagagg
     3121 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     3181 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3241 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3301 tcatcaatgt atcaggcctt ggtaccgttt aaacttaaaa tgcatgcggc cgccaccgcg
     3361 gtggagctcc agcttttgtt ccctttagtg agggttaatt gggcccaagt gatctccaaa
     3421 aaataagtac tttttgactg taaataaaat tgtaaggagt aaaaagtact tttttttcta
     3481 aaaaaatgta attaagtaaa agtaaaagta ttgattttta attgtactca agtaaagtaa
     3541 aaatccccaa aaataatact taagtacagt aatcaagtaa aattactcaa gtactttaca
     3601 cctctgggta ctgtttcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3661 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     3721 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3781 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3841 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3901 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3961 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4021 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4081 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4141 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4201 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4261 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4321 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4381 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4441 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4501 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4561 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4621 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4681 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4741 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4801 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4861 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4921 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4981 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5041 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5101 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5161 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5221 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5281 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5341 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5401 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5461 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5521 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5581 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5641 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5701 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5761 cgcacatttc cccgaaaagt gccac
//
