LOCUS       sequence-36625-e        5207 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5207)
  AUTHORS   Petersen LK, Stowers RS
  TITLE     A Gateway MultiSite Recombination Cloning Toolkit.
  JOURNAL   PLoS One. 2011;6(9):e24531. Epub 2011 Sep 9.
  PUBMED    21931740
REFERENCE   2  (bases 1 to 5207)
  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: "2011"; volume: "6(9)"; pages: "e24531. Epub 2011 Sep 9"
COMMENT     Sequence Label: pENTR L5-GAL4-L2
FEATURES             Location/Qualifiers
     source          1..5207
                     /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    570..665
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             672..3317
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4"
                     /product="GAL4 transcriptional activator"
                     /label=GAL4
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVSIDSAAHHDNSTIPLDF
                     MPRDALHGFDWSEEDDMSDGLPFLKTDPNNNGFFGDGSLLCILRSIGFKPENYTNSNVN
                     RLPTMITDRYTLASRSTTSRLLQSYLNNFHPYCPIVHSPTLMMLYNNQIEIASKDQWQI
                     LFNCILAIGAWCIEGESTDIDVFYYQNAKSHLTSKVFESGSIILVTALHLLSRYTQWRQ
                     KTNTSYNFHSFSIRMAISLGLNRDLPSSFSDSSILEQRRRIWWSVYSWEIQLSLLYGRS
                     IQLSQNTISFPSSVDDVQRTTTGPTIYHGIIETARLLQVFTKIYELDKTVTAEKSPICA
                     KKCLMICNEIEEVSRQAPKFLQMDISTTALTNLLKEHPWLSFTRFELKWKQLSLIIYVL
                     RDFFTNFTQKKSQLEQDQNDHQSYEVKRCSIMLSDAAQRTVMSVSSYMDNHNVTPYFAW
                     NCSYYLFNAVLVPIKTLLSNSKSNAENNETAQLLQQINTVLMLLKKLATFKIQTCEKYI
                     QVLEEVCAPFLLSQCAIPLPHISYNNSNGSAIKNIVGSATIAQYPTLPEENVNNISVKY
                     VSPGSVGPSPVPLKSGASFSDLVKLLSNRPPSRNSPVTIPRSTPSHRSVTPFLGQQQQL
                     QSLVPLTPSALFGGANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     protein_bind    complement(3319..3418)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3448..3467)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3449..3467)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3472..3488)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3472..3488)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3590..3609)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3601..4410
                     /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(3668..3687)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4557..5145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5046..5065
                     /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 tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtatacaaa
      661 agttggccaa catgaagcta ctgtcttcta tcgaacaagc atgcgatatt tgccgactta
      721 aaaagctcaa gtgctccaaa gaaaaaccga agtgcgccaa gtgtctgaag aacaactggg
      781 agtgtcgcta ctctcccaaa accaaaaggt ctccgctgac tagggcacat ctgacagaag
      841 tggaatcaag gctagaaaga ctggaacagc tatttctact gatttttcct cgagaagacc
      901 ttgacatgat tttgaaaatg gattctttac aggatataaa agcattgtta acaggattat
      961 ttgtacaaga taatgtgaat aaagatgccg tcacagatag attggcttca gtggagactg
     1021 atatgcctct aacattgaga cagcatagaa taagtgcgac atcatcatcg gaagagagta
     1081 gtaacaaagg tcaaagacag ttgactgtat cgattgactc ggcagctcat catgataact
     1141 ccacaattcc gttggatttt atgcccaggg atgctcttca tggatttgat tggtctgaag
     1201 aggatgacat gtcggatggc ttgcccttcc tgaaaacgga ccccaacaat aatgggttct
     1261 ttggcgacgg ttctctctta tgtattcttc gatctattgg ctttaaaccg gaaaattaca
     1321 cgaactctaa cgttaacagg ctcccgacca tgattacgga tagatacacg ttggcttcta
     1381 gatccacaac atcccgttta cttcaaagtt atctcaataa ttttcacccc tactgcccta
     1441 tcgtgcactc accgacgcta atgatgttgt ataataacca gattgaaatc gcgtcgaagg
     1501 atcaatggca aatccttttt aactgcatat tagccattgg agcctggtgt atagaggggg
     1561 aatctactga tatagatgtt ttttactatc aaaatgctaa atctcatttg acgagcaagg
     1621 tcttcgagtc aggttccata attttggtga cagccctaca tcttctgtcg cgatatacac
     1681 agtggaggca gaaaacaaat actagctata attttcacag cttttccata agaatggcca
     1741 tatcattggg cttgaatagg gacctcccct cgtccttcag tgatagcagc attctggaac
     1801 aaagacgccg aatttggtgg tctgtctact cttgggagat ccaattgtcc ctgctttatg
     1861 gtcgatccat ccagctttct cagaatacaa tctccttccc ttcttctgtc gacgatgtgc
     1921 agcgtaccac aacaggtccc accatatatc atggcatcat tgaaacagca aggctcttac
     1981 aagttttcac aaaaatctat gaactagaca aaacagtaac tgcagaaaaa agtcctatat
     2041 gtgcaaaaaa atgcttgatg atttgtaatg agattgagga ggtttcgaga caggcaccaa
     2101 agtttttaca aatggatatt tccaccaccg ctctaaccaa tttgttgaag gaacaccctt
     2161 ggctatcctt tacaagattc gaactgaagt ggaaacagtt gtctcttatc atttatgtat
     2221 taagagattt tttcactaat tttacccaga aaaagtcaca actagaacag gatcaaaatg
     2281 atcatcaaag ttatgaagtt aaacgatgct ccatcatgtt aagcgatgca gcacaaagaa
     2341 ctgttatgtc tgtaagtagc tatatggaca atcataatgt caccccatat tttgcctgga
     2401 attgttctta ttacttgttc aatgcagtcc tagtacccat aaagactcta ctctcaaact
     2461 caaaatcgaa tgctgagaat aacgagaccg cacaattatt acaacaaatt aacactgttc
     2521 tgatgctatt aaaaaaactg gccactttta aaatccagac ttgtgaaaaa tacattcaag
     2581 tactggaaga ggtatgtgcg ccgtttctgt tatcacagtg tgcaatccca ttaccgcata
     2641 tcagttataa caatagtaat ggtagcgcca ttaaaaatat tgtcggttct gcaactatcg
     2701 cccaataccc tactcttccg gaggaaaatg tcaacaatat cagtgttaaa tatgtttctc
     2761 ctggctcagt agggccttca cctgtgccat tgaaatcagg agcaagtttc agtgatctag
     2821 tcaagctgtt atctaaccgt ccaccctctc gtaactctcc agtgacaata ccaagaagca
     2881 caccttcgca tcgctcagtc acgccttttc tagggcaaca gcaacagctg caatcattag
     2941 tgccactgac cccgtctgct ttgtttggtg gcgccaattt taatcaaagt gggaatattg
     3001 ctgatagctc attgtccttc actttcacta acagtagcaa cggtccgaac ctcataacaa
     3061 ctcaaacaaa ttctcaagcg ctttcacaac caattgcctc ctctaacgtt catgataact
     3121 tcatgaataa tgaaatcacg gctagtaaaa ttgatgatgg taataattca aaaccactgt
     3181 cacctggttg gacggaccaa actgcgtata acgcgtttgg aatcactaca gggatgttta
     3241 ataccactac aatggatgat gtatataact atctattcga tgatgaagat accccaccaa
     3301 acccaaaaaa agagtaatac ccagctttct tgtacaaagt tggcattata agaaagcatt
     3361 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc
     3421 atccagctgc agggcggccg cgatatcccc tatagtgagt cgtattacat ggtcatagct
     3481 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     3541 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     3601 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     3661 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     3721 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     3781 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     3841 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     3901 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     3961 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     4021 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     4081 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     4141 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     4201 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     4261 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     4321 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     4381 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     4441 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     4501 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4561 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4621 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4681 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4741 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4801 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4861 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4921 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4981 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5041 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5101 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5161 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
