LOCUS       sequence-64547-e        2955 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 2955)
  AUTHORS   Giry-Laterriere M, Cherpin O, Kim YS, Jensen J, Salmon P
  TITLE     Polyswitch lentivectors: ""all-in-one"" lentiviral vectors for 
            drug-inducible gene expression, live selection, and recombination 
            cloning.
  JOURNAL   Hum Gene Ther. 2011 Oct;22(10):1255-67. Epub 2011 Sep 27.
  PUBMED    21761975
REFERENCE   2  (bases 1 to 2955)
  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: "Hum Gene 
            Ther."; date: "2011-10"; volume: "22(10)"; pages: "1255-67. Epub 
            2011 Sep 27"
COMMENT     Sequence Label: pENTR-L4-pTF-L1R
FEATURES             Location/Qualifiers
     source          1..2955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..28)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(120..206)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     255..277
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     269..286
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     270..286
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    326..421
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    448..466
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    489..507
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    653..671
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    734..857
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(929..948)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(930..948)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(953..969)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(953..969)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1071..1090)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1082..1891
                     /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(1149..1168)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2038..2626
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2527..2546
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2780..2797
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg
       61 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga
      121 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
      181 tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct
      241 agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc
      301 cgcgttaacg ctaccatgga gctccaaata atgattttat tttgactgat agtgacctgt
      361 tcgttgcaac aaattgataa gcaatgcttt tttataatgc caactttgta tagaaaagtt
      421 gggtacaact cccactagac atatcctact ctatcattga tagagtgacc cttccataga
      481 catatcctac tctatcattg atagagtgac ccttccacta ggttcacttt tctctatcat
      541 tgatagggag tggtaaacaa ttgatggaca aaatctatca atgatagaga gtgcatgtta
      601 ctagtattgt tcttctctat cattgatagt gactttgcct aatcacttag caactctatc
      661 attgatagag ttctagtatt agcaaggcct ttcctctata taaggaagtt catttcattt
      721 ggagaagatc ccccaagttt gtacaaaaaa gttgaacgag aaacgtaaaa tgatataaat
      781 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
      841 catatgcagt cactatgaat caactactta gatggtatta gtgacctgta gaattcgagc
      901 tctagagctg cagggcggcc gcgatatccc ctatagtgag tcgtattaca tggtcatagc
      961 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     1021 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     1081 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     1141 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     1201 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     1261 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     1321 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     1381 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     1441 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     1501 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     1561 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     1621 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     1681 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     1741 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     1801 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     1861 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     1921 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     1981 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2041 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2101 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2161 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     2221 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2281 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2341 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2401 caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag
     2461 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2521 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2581 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2641 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2701 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2761 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     2821 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     2881 cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga
     2941 gtttgtagaa acgca
//
