LOCUS       sequence-161165-        3920 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' entry vector containing frog translation elongation factor 1 
            alpha (EF1a) enhancer fused to rabbit b-globin intron for 
            semi-ubiquitous expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3920)
  AUTHORS   Akerberg AA, Stewart S, Stankunas K
  TITLE     Spatial and temporal control of transgene expression in zebrafish.
  JOURNAL   PLoS One. 2014 Mar 18;9(3):e92217. doi: 
            10.1371/journal.pone.0092217. eCollection 2014.
  PUBMED    24643048
REFERENCE   2  (bases 1 to 3920)
  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: "2014-03-18"; volume: "9(3):e92217. doi"; pages: " 
            10.1371/journal.pone.0092217. eCollection 2014"
COMMENT     Sequence Label: p5E-EF1a/b-globin
FEATURES             Location/Qualifiers
     source          1..3920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    593..688
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        690..708
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          1260..1832
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(1894..1910)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1894..1910)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1947..1965)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    1966..2089
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2162..2180)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2185..2201)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2185..2201)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2303..2322)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2314..3123
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2381..2400)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3270..3858
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3759..3778
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccgc gttaacgcta ccatggagct ccaaataatg
      601 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca atgctttttt
      661 ataatgccaa ctttgtatag aaaagttggt aatacgactc actatagggc gaattgggta
      721 ccgggccccc cctcgaggtc gagcaggggg atcatctaat caagcacaaa taaggggcgt
      781 gtaacacaaa agccagcgac cctttccaat gcaaatcaaa cttgcaattc tttgccgttt
      841 ttatcattta agtgtcggct taaggtccac tatcagatgt aaacagcctt atctaacaaa
      901 ggtatcatta cattctgaaa ttctcaggca tgcaagctag cttatgacgc actagggagt
      961 gccacccttc ctttcgccct aacttcgtga taactcgcgc gtttcactca acagctgcat
     1021 ccgccctagt gctactggga gttgtagtat acaagacgct tacaggctga atgttctgtc
     1081 aagaccccgc ctctagcact ttgggaattc tggacttgat gatgtcatgg ttaatccccg
     1141 cccagtagag gcggctatat aaagggtggt taaggcccgg ttcgctctct tcctcaccgg
     1201 gtctgcggcg agttctagct gaagcttcct gcaggtcgac cgatcctgag aacttcaggg
     1261 tgagtttggg gacccttgat tgttctttct ttttcgctat tgtaaaattc atgttatatg
     1321 gagggggcaa agttttcagg gtgttgttta gaatgggaag atgtcccttg tatcaccatg
     1381 gaccctcatg ataattttgt ttctttcact ttctactctg ttgacaacca ttgtctcctc
     1441 ttattttctt ttcattttct gtaacttttt cgttaaactt tagcttgcat ttgtaacgaa
     1501 tttttaaatt cacttttgtt tatttgtcag attgtaagta ctttctctaa tcactttttt
     1561 ttcaaggcaa tcagggtata ttatattgta cttcagcaca gttttagaga acaattgtta
     1621 taattaaatg ataaggtaga atatttctgc atataaattc tggctggcgt ggaaatattc
     1681 ttattggtag aaacaactac accctggtca tcatcctgcc tttctcttta tggttacaat
     1741 gatatacact gtttgagatg aggataaaat actctgagtc caaaccgggc ccctctgcta
     1801 accatgttca tgccttcttc tctttcctac agctcctggg caacgtgctg gttgttgtgc
     1861 tgtctcatca ttttggcaaa gaattcctcg acggatccac tagttctaga gcggccgcca
     1921 ccgcggtgga gctccagctt ttgttccctt tagtgagggt taattcaagt ttgtacaaaa
     1981 aagttgaacg agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata
     2041 aaaaacagac tacataatac tgtaaaacac aacatatgca gtcactatga accaactact
     2101 tagatggtat tagtgacctg tagaattcga gctctagagc tgcagggcgg ccgcgatatc
     2161 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     2221 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     2281 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
     2341 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     2401 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
     2461 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     2521 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     2581 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
     2641 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     2701 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     2761 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     2821 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     2881 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     2941 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     3001 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     3061 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     3121 taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac
     3181 ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     3241 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3301 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3361 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     3421 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3481 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3541 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3601 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3661 cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3721 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3781 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3841 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3901 tggccttttg ctcacatgtt
//
