LOCUS       sequence-447206-        3148 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway compatible 5' entry clone containing a 3rd generation Tet-On
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3148)
  AUTHORS   Gillespie WB 3rd, Zhang Y, Ruiz OE, Cerda J 3rd, Ortiz-Guzman J, 
            Sherman M, Turner WD, Largoza G, Mosser LE, Fujimoto E, Chien CB, 
            Kwan KM, Arenkiel BR, Devine WP, Wythe JD
  TITLE     MultiSite Assembly of Gateway Induced Clones (MAGIC): a flexible 
            cloning toolbox for use in vertebrate model systems.
  JOURNAL   Development. 2025 Nov 15;152(22):dev204308. doi: 10.1242/dev.204308.
            Epub 2025 Nov 13.
  PUBMED    41230856
REFERENCE   2  (bases 1 to 3148)
  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: 
            "Development."; date: "2025-11-15"; volume: "152(22):dev204308. 
            doi"; pages: " 10.1242/dev.204308. Epub 2025 Nov 13"
COMMENT     Sequence Label: p5E TRE-3GV (JDW 1163)
FEATURES             Location/Qualifiers
     source          1..3148
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     73..90
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(249..276)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(368..454)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     503..525
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     517..534
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     518..534
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    574..669
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        671..689
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        716..1065
                     /label=TRE3GV promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, optimized for 
                     retroviral and lentiviral vectors"
     protein_bind    724..742
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    760..778
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    796..814
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    832..850
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    868..886
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    904..922
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    940..958
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1004..1028
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(1103..1119)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1103..1119)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1156..1174)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    1175..1298
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1371..1389)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1394..1410)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1394..1410)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1512..1531)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1523..2332
                     /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(1590..1609)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2479..3067
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2968..2987
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
       61 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
      121 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
      181 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta
      241 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
      301 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
      361 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
      421 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
      481 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag
      541 ctcgggcccg agttaacgct accatggagc tccaaataat gattttattt tgactgatag
      601 tgacctgttc gttgcaacaa attgataagc aatgcttttt tataatgcca actttgtata
      661 gaaaagttgg taatacgact cactataggg cgaattgggt accgggcccc ccctcgagtt
      721 tactccctat cagtgataga gaacgtatga agagtttact ccctatcagt gatagagaac
      781 gtatgcagac tttactccct atcagtgata gagaacgtat aaggagttta ctccctatca
      841 gtgatagaga acgtatgacc agtttactcc ctatcagtga tagagaacgt atctacagtt
      901 tactccctat cagtgataga gaacgtatat ccagtttact ccctatcagt gatagagaac
      961 gtatgtcgag gtaggcgtgt acggtgggcg cctataaaag cagagctcgt ttagtgaacc
     1021 gtcagatcgc ctggagcaat tccacaacac ttttgtctta tacttacgcg taagcttgat
     1081 atcgaattcc tgcagcccgg gggatccact agttctagag cggccgccac cgcggtggag
     1141 ctccagcttt tgttcccttt agtgagggtt aattcaagtt tgtacaaaaa agttgaacga
     1201 gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact
     1261 acataatact gtaaaacaca acatatgcag tcactatgaa ccaactactt agatggtatt
     1321 agtgacctgt agaattcgag ctctagagct gcagggcggc cgcgatatcc cctatagtga
     1381 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     1441 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     1501 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     1561 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     1621 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     1681 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     1741 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     1801 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     1861 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     1921 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     1981 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     2041 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     2101 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     2161 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     2221 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     2281 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     2341 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     2401 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     2461 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2521 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2581 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2641 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2701 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2761 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2821 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2881 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2941 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3001 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3061 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3121 tcacatgttc tttcctgcgt tatcccct
//
