LOCUS       sequence-346784-        4014 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 4014)
  AUTHORS   Korneel Hens, Jean-Daniel Feuz, Alina Isakova, Antonina Lagovitina, 
            Andreas Massouras, Julien Bryois, Patrick Callaerts, Susan E. 
            Celniker, Bart Deplancke
  TITLE     Automated protein-DNA interaction screening of Drosophila regulatory
            elements
  JOURNAL   Nat Methods. 2011 Oct 30;8(12):1065-70.
  PUBMED    22037703
REFERENCE   2  (bases 1 to 4014)
  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: "Nat 
            Methods."; date: "2011-10-30"; volume: "8(12)"; pages: "1065-70"
COMMENT     Sequence Label: KHBD00188
FEATURES             Location/Qualifiers
     source          1..4014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1603..1702)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1719..1738)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1720..1738)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1743..1759)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1743..1759)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1861..1880)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1872..2681
                     /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(1939..1958)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2828..3416
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3317..3336
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3570..3587
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3746..3773)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3865..3951)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4000..4014,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4014,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagttgg caccatggcg caaccgccgc cagatcaaaa cttttaccat catccgctgc
      181 cccacacgca cacacatccg catccgcact cccatccgca tccgcactcg catccgcacc
      241 cacatcacca acatccgcag cttcagttgc cgccacaatt ccgaaatccc ttcgatttgc
      301 ttttcgatga gcgaacggga gcgataaact acaactacat acgtccgtat ctgcccaacc
      361 agatgcccaa gccagaggag ctgcccgact ctctggtgat gcggcgacca cgtcgcaccc
      421 gcaccacttt taccagctct caaatagcag agctggagca gcactttctg cagggacgat
      481 acctcacagc cccccgactt gcggatctgt cagcgaaact agccctgggc acagcccagg
      541 tgaagatatg gtttaagaac cgtcggcgtc gtcacaagat ccaatcggat cagcacaagg
      601 accagtccta cgaggggatg cctctctcgc cgggtatgaa acagagcgat ggcgatcccc
      661 ccagcttgca gactcttagc ttgggtggag gagccacgcc caacgctttg actccgtcac
      721 ccacgccctc aacgcccact gcacacatga cggagcacta cagcgagtca ttcaacgcct
      781 actacaacta caatggaggc cacaatcacg cccaggccaa tcgtcacatg cacatgcagt
      841 atccttccgg aggggggcca ggacctgggt cgaccaatgt caatggcggc cagttcttcc
      901 agcagcagca ggtccataat caccagcagc aactgcacca ccagggcaac cacgtgccgc
      961 accagatgca gcagcagcaa cagcaggctc agcagcagca ataccatcac tttgacttcc
     1021 agcaaaagca agccagcgcc tgtcgcgtcc tggtcaagga cgaaccggag gccgactaca
     1081 acttcaacag ctcgtactac atgcgatcgg gaatgtctgg cgccactgca tcggcatccg
     1141 ctgtggcccg aggcgctgcc tcgccgggct ccgaggtcta cgagccatta acacccaaga
     1201 atgacgaaag tccgagtctg tgtggcatcg gcatcggcgg accttgcgcc atcgccgttg
     1261 gcgagacgga ggcggccgac gacatggacg acggaacgag caagaagacg acgctacaga
     1321 tcttggagcc tttgaagggt ctggacaaga gctgcgacga tggcagtagc gacgacatga
     1381 gcaccggaat aagagcctta gcaggaaccg gaaatcgtgg agcggcattt gccaaatttg
     1441 gcaagccttc gcccccacaa ggccctcagc cgcccctcgg aatggggggc gtggccatgg
     1501 gcgaatcgaa ccaatatcaa tgcacgatgg atacgataat gcaagcgtat aatccccatc
     1561 ggaacgccgc gggcaactcg cagtttgcct actgcttcaa tttgccaact ttcttgtaca
     1621 aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca
     1681 gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca
     1741 tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg
     1801 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     1861 caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca
     1921 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     1981 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     2041 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     2101 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     2161 ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     2221 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     2281 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     2341 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     2401 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     2461 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     2521 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     2581 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     2641 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt
     2701 tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat
     2761 cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     2821 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     2881 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     2941 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3001 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3061 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3121 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3181 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3241 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3301 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3361 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3421 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     3481 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     3541 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     3601 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     3661 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta
     3721 gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt
     3781 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac
     3841 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca
     3901 acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc
     3961 agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgtt
//
