LOCUS       sequence-101786-        4632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mRNA synthesis of Gal4ff codon modified for zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4632)
  AUTHORS   Horstick EJ, Jordan DC, Bergeron SA, Tabor KM, Serpe M, Feldman B, 
            Burgess HA
  TITLE     Increased functional protein expression using nucleotide sequence 
            features enriched in highly expressed genes in zebrafish.
  JOURNAL   Nucleic Acids Res. 2015 Jan 27. pii: gkv035.
  PUBMED    25628360
REFERENCE   2  (bases 1 to 4632)
  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: "Nucleic 
            Acids Res. 2015 Jan 27. pii: gkv035."
COMMENT     Sequence Label: pCS2-Gal4ff.zf1
FEATURES             Location/Qualifiers
     source          1..4632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             91..531
                     /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"
     polyA_signal    681..815
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(705..724)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     759..778
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(928..948)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(928..946)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(967..983)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(967..983)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(980..1002)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    991..1007
                     /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(1015..1021,1022..1039,1040..1045))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1060..1081
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1198..1215)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1369..1957)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1449..1468)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2128..2919,2920..2988))
                     /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     2751..2770
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2989..3093)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3119..3574)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3256..3277)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3468..3487
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3648..4632
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccgccacc atgaaactgc tgagctccat tgaacaggcc
      121 tgtgatatct gcaggctgaa gaagctgaag tgtagcaagg agaagcctaa gtgtgctaag
      181 tgtctgaaga acaactggga gtgtagatac agccctaaga caaagagaag ccctctgaca
      241 agggctcacc tgacagaggt ggagagcaga ctggagagac tggagcagct gttcctgctg
      301 atcttcccta gagaggacct ggacatgatc ctgaagatgg acagcctcca ggacatcaag
      361 gctctgctga caggactgtt tgtgcaggac aatgtgaaca aggatgctgt gacagacaga
      421 ctggcttctg tggagacaga catgcctctg acactgagac agcacagaat ctctgctaca
      481 agcagctctg aggagagcag caacaaggga cagagacagc tgacagtgag ccctgagttc
      541 cctgctgatg ctctggatga ctttgacctg gacatgctgc ctgctgatgc tctggatgac
      601 tttgacctgg acatgctgcc tggaaactaa aaattcaagg cctctcgagc ctctagaact
      661 atagtgagtc gtattacgta gatccagaca tgataagata cattgatgag tttggacaaa
      721 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
      781 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
      841 tgtttcaggt tcagggggag gtgtgggagg ttttttaatt cgcggccgcg gcgccaatgc
      901 attgggcccg gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
      961 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     1021 acgagccggg agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     1081 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     1141 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     1201 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1261 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1321 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1381 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1441 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1501 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1561 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1621 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1681 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1741 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1801 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1861 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1921 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1981 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2041 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2101 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2161 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     2221 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     2281 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     2341 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     2401 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     2461 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     2521 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     2581 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     2641 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     2701 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     2761 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     2821 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     2881 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     2941 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3001 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3061 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     3121 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     3181 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     3241 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     3301 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     3361 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3421 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3481 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3541 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
     3601 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagtcgac catagccaat
     3661 tcaatatggc gtatatggac tcatgccaat tcaatatggt ggatctggac ctgtgccaat
     3721 tcaatatggc gtatatggac tcgtgccaat tcaatatggt ggatctggac cccagccaat
     3781 tcaatatggc ggacttggca ccatgccaat tcaatatggc ggacttggca ctgtgccaac
     3841 tggggagggg tctacttggc acggtgccaa gtttgaggag gggtcttggc cctgtgccaa
     3901 gtccgccata ttgaattggc atggtgccaa taatggcggc catattggct atatgccagg
     3961 atcaatatat aggcaatatc caatatggcc ctatgccaat atggctattg gccaggttca
     4021 atactatgta ttggccctat gccatatagt attccatata tgggttttcc tattgacgta
     4081 gatagcccct cccaatgggc ggtcccatat accatatatg gggcttccta ataccgccca
     4141 tagccactcc cccattgacg tcaatggtct ctatatatgg tctttcctat tgacgtcata
     4201 tgggcggtcc tattgacgta tatggcgcct cccccattga cgtcaattac ggtaaatggc
     4261 ccgcctggct caatgcccat tgacgtcaat aggaccaccc accattgacg tcaatgggat
     4321 ggctcattgc ccattcatat ccgttctcac gccccctatt gacgtcaatg acggtaaatg
     4381 gcccacttgg cagtacatca atatctatta atagtaactt ggcaagtaca ttactattgg
     4441 aaggacgcca gggtacattg gcagtactcc cattgacgtc aatggcggta aatggcccgc
     4501 gatggctgcc aagtacatcc ccattgacgt caatggggag gggcaatgac gcaaatgggc
     4561 gttccattga cgtaaatggg cggtaggcgt gcctaatggg aggtctatat aagcaatgct
     4621 cgtttaggga ac
//
