LOCUS       sequence-36629-e        5317 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 5317)
  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 5317)
  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 R4-GAL4-R3
FEATURES             Location/Qualifiers
     source          1..5317
                     /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    complement(603..727)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             733..3378
                     /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    3380..3503
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3558..3577)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3559..3577)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3582..3598)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3582..3598)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3700..3719)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3711..4520
                     /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(3778..3797)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4667..5255
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5156..5175
                     /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 tcgggcccct acaggtcact aataccatct aagtagttga
      601 ttcatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
      661 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact tttctataca
      721 aagttggcca acatgaagct actgtcttct atcgaacaag catgcgatat ttgccgactt
      781 aaaaagctca agtgctccaa agaaaaaccg aagtgcgcca agtgtctgaa gaacaactgg
      841 gagtgtcgct actctcccaa aaccaaaagg tctccgctga ctagggcaca tctgacagaa
      901 gtggaatcaa ggctagaaag actggaacag ctatttctac tgatttttcc tcgagaagac
      961 cttgacatga ttttgaaaat ggattcttta caggatataa aagcattgtt aacaggatta
     1021 tttgtacaag ataatgtgaa taaagatgcc gtcacagata gattggcttc agtggagact
     1081 gatatgcctc taacattgag acagcataga ataagtgcga catcatcatc ggaagagagt
     1141 agtaacaaag gtcaaagaca gttgactgta tcgattgact cggcagctca tcatgataac
     1201 tccacaattc cgttggattt tatgcccagg gatgctcttc atggatttga ttggtctgaa
     1261 gaggatgaca tgtcggatgg cttgcccttc ctgaaaacgg accccaacaa taatgggttc
     1321 tttggcgacg gttctctctt atgtattctt cgatctattg gctttaaacc ggaaaattac
     1381 acgaactcta acgttaacag gctcccgacc atgattacgg atagatacac gttggcttct
     1441 agatccacaa catcccgttt acttcaaagt tatctcaata attttcaccc ctactgccct
     1501 atcgtgcact caccgacgct aatgatgttg tataataacc agattgaaat cgcgtcgaag
     1561 gatcaatggc aaatcctttt taactgcata ttagccattg gagcctggtg tatagagggg
     1621 gaatctactg atatagatgt tttttactat caaaatgcta aatctcattt gacgagcaag
     1681 gtcttcgagt caggttccat aattttggtg acagccctac atcttctgtc gcgatataca
     1741 cagtggaggc agaaaacaaa tactagctat aattttcaca gcttttccat aagaatggcc
     1801 atatcattgg gcttgaatag ggacctcccc tcgtccttca gtgatagcag cattctggaa
     1861 caaagacgcc gaatttggtg gtctgtctac tcttgggaga tccaattgtc cctgctttat
     1921 ggtcgatcca tccagctttc tcagaataca atctccttcc cttcttctgt cgacgatgtg
     1981 cagcgtacca caacaggtcc caccatatat catggcatca ttgaaacagc aaggctctta
     2041 caagttttca caaaaatcta tgaactagac aaaacagtaa ctgcagaaaa aagtcctata
     2101 tgtgcaaaaa aatgcttgat gatttgtaat gagattgagg aggtttcgag acaggcacca
     2161 aagtttttac aaatggatat ttccaccacc gctctaacca atttgttgaa ggaacaccct
     2221 tggctatcct ttacaagatt cgaactgaag tggaaacagt tgtctcttat catttatgta
     2281 ttaagagatt ttttcactaa ttttacccag aaaaagtcac aactagaaca ggatcaaaat
     2341 gatcatcaaa gttatgaagt taaacgatgc tccatcatgt taagcgatgc agcacaaaga
     2401 actgttatgt ctgtaagtag ctatatggac aatcataatg tcaccccata ttttgcctgg
     2461 aattgttctt attacttgtt caatgcagtc ctagtaccca taaagactct actctcaaac
     2521 tcaaaatcga atgctgagaa taacgagacc gcacaattat tacaacaaat taacactgtt
     2581 ctgatgctat taaaaaaact ggccactttt aaaatccaga cttgtgaaaa atacattcaa
     2641 gtactggaag aggtatgtgc gccgtttctg ttatcacagt gtgcaatccc attaccgcat
     2701 atcagttata acaatagtaa tggtagcgcc attaaaaata ttgtcggttc tgcaactatc
     2761 gcccaatacc ctactcttcc ggaggaaaat gtcaacaata tcagtgttaa atatgtttct
     2821 cctggctcag tagggccttc acctgtgcca ttgaaatcag gagcaagttt cagtgatcta
     2881 gtcaagctgt tatctaaccg tccaccctct cgtaactctc cagtgacaat accaagaagc
     2941 acaccttcgc atcgctcagt cacgcctttt ctagggcaac agcaacagct gcaatcatta
     3001 gtgccactga ccccgtctgc tttgtttggt ggcgccaatt ttaatcaaag tgggaatatt
     3061 gctgatagct cattgtcctt cactttcact aacagtagca acggtccgaa cctcataaca
     3121 actcaaacaa attctcaagc gctttcacaa ccaattgcct cctctaacgt tcatgataac
     3181 ttcatgaata atgaaatcac ggctagtaaa attgatgatg gtaataattc aaaaccactg
     3241 tcacctggtt ggacggacca aactgcgtat aacgcgtttg gaatcactac agggatgttt
     3301 aataccacta caatggatga tgtatataac tatctattcg atgatgaaga taccccacca
     3361 aacccaaaaa aagagtaaac aactttgtat aataaagttg aacgagaaac gtaaaatgat
     3421 ataaatatca atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa
     3481 acacaacata tccagtcact atgaatcaac tacttagatg gtattagtga cctgtactgc
     3541 agggcggccg cgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc
     3601 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
     3661 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
     3721 ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     3781 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga
     3841 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     3901 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     3961 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag
     4021 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     4081 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     4141 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     4201 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
     4261 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     4321 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     4381 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     4441 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     4501 tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta
     4561 cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt
     4621 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4681 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4741 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4801 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4861 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4921 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4981 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5041 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5101 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5161 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5221 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5281 ggttcctggc cttttgctgg ccttttgctc acatgtt
//
