LOCUS       sequence-99768-e        5723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Middle Gateway Entry Vector Containing QF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5723)
  AUTHORS   Subedi A, Macurak M, Gee ST, Monge E, Goll MG, Potter CJ, Parsons 
            MJ, Halpern ME
  TITLE     Adoption of the Q transcriptional regulatory system for zebrafish 
            transgenesis.
  JOURNAL   Methods. 2014 Apr 1;66(3):433-40. doi: 10.1016/j.ymeth.2013.06.012. 
            Epub 2013 Jun 20.
  PUBMED    23792917
REFERENCE   2  (bases 1 to 5723)
  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"; doi: 
            "10.1016/j.ymeth.2013.06.012"; journalName: "Methods"; date: 
            "2014-04-1- 1"; volume: "66"; issue: "3"; pages: "433-40"
COMMENT     Sequence Label: pME-QF
FEATURES             Location/Qualifiers
     source          1..5723
                     /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    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     intron          3214..3279
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3409..3429
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3701..3835
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3738..3757)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3792..3811
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3848..3947)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3964..3983)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3965..3983)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3988..4004)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3988..4004)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4106..4125)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4117..4926
                     /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(4184..4203)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5073..5661
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5562..5581
                     /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 tgtacaaaaa
      661 agcaggcttt ccaacatgcc gcctaaacgc aagacactca atgccgctgc cgaagccaat
      721 gcccacgctg atggccatgc tgatggcaat gctgatggtc atgtcgctaa cactgcagca
      781 agcagcaaca acgcccgttt tgcggacttg accaacattg acacacccgg cctcggccct
      841 accacgacga cgttacttgt cgagcccgct cgttcgaaac gccagagagt ctcgagggcc
      901 tgtgatcagt gtcgagctgc acgtgaaaag tgtgatggaa tccagccggc ttgcttcccc
      961 tgtgtgtcgc agggccggtc gtgtacctac caggccagtc ccaagaagcg aggagtccag
     1021 acgggctaca tccgcactct cgaactggct ctggcttgga tgttcgagaa cgttgcccgc
     1081 agcgaggacg ccctccacaa tcttttggtc cgtgatgctg gccagggcag cgctctcctg
     1141 gtcggcaaag actcgcctgc tgcagaacgc ctgcatgcaa gatgggcgac gagtcgagtc
     1201 aacaaaagca tcacccgtct tctctcaggt caggccgcac aagatccatc tgaagacggc
     1261 caatccccgt ccgaagacat aaatgtccaa gatgccgggg caaagacatc cgacttccct
     1321 catgcgcctc acttgacttt ctcggcgcct aagtctagta cagctgagac acgcactcta
     1381 ccaggcccgg tccgaccacc tatttcggca aacaccctgg aaaacaacct tcagccagat
     1441 ggtaccggga tagggaagct accacccaat cattggcgcc tgctggatat ctacttttcc
     1501 tacacgcatt cttggctccc tatcctcgag aagaaagaca tgtaccaagc attgtaccag
     1561 tactctgaac aaggctcgtt gcttccctct gcgaatgtcg agtctggcgt tcatgccgag
     1621 ctctggagcg cgctcgccct ggcgtccttc caggctgctg ctactgctgc atcgagtgct
     1681 acgggtccag cttcagctgc tcatggccat gacaatgcca tcaatccttc acctgcagac
     1741 atatctgaca cagcccgaaa gctcatacct ttggaaagcg ggccgttcca ggttcagcac
     1801 tgcagagcgt tgctgcttct ttgtctcgta agccttgggc gggatgattg ggagtctgct
     1861 tggttgctgg ttggctttgc ggtccgcgtc ctacttgttg ttcgcaccca gttgcctcct
     1921 gatgatgacc ggccacgacc aagaatgcgt gcgctgctcg tcgcgtgctt catcgtggat
     1981 accattgtgt ctatgagaca caacgtgccg gcccatctca agccagacga cattgcggat
     2041 ctgccgttac ctgaagacgg tcaagatcaa tgggagccgt ggacaccatg tgagggctta
     2101 ggcggtgaac acaccatgct gcaaatgttg aggaacccgg catacccttt aagcacattc
     2161 aaccacctat atggcgtgac caagctggtt gctttggagc ttctgccaag aatacgaaca
     2221 tcttcacaga acgctccctt ggagttcagg tcgcggttgc agcaggtaat cggccacaat
     2281 tctcccttca gcgtctttgt cctttcccag gatacagcat cggcttttgt gcctactgca
     2341 taccttaccc gtaccgttta tttatgggca gctgcctttt ctgagcctct caacgaacac
     2401 tactcgcatc ttctgatcga aactcttgat cagtatcaga agcggtttgg tacatatgca
     2461 atcccacctc tgatcccttc tcttctagac tcccttcttg ctttaaagaa acaatcacat
     2521 tcttcagagc ggcatcgaag gcacttggaa gagcttttcc ccgcctactc ctccatttgg
     2581 cctcggggag gccgacacag caatactggc ctccaaccca tacgacaact tgagcttccg
     2641 ccgactgcga ctgccactgc aagtatcatg ccccatgtca tggaacagcc cctgtcaacg
     2701 tcaataaatc cggtcaatga tcggtttaat ggaataccga atcctacccc ctacaatagc
     2761 gatgcggccc tagacgcaat tactcaaacc aatgactacg gatcagtcaa tacccatggc
     2821 attctcagta catatccacc acctgccacc cacttgaacg aggcctcggt ggcccttgcg
     2881 cctggaggtg ctcctcccag accacccccg ccatacgttg atagtacaac gaaccatcct
     2941 ccttaccata gtaacctcgt ccccatggcc aatttcggat attcgactgt agactatgat
     3001 gcgatggtgg atgatctggc atcgatcgag tacacggatg cggttgacgt tgatccgcag
     3061 ttcatgacga acctcgggtt tgtcccaggg tgtaatttta gcgatatcaa cacgtatgag
     3121 caataggacg tctagaggat ctttgtgaag gaaccttact tctgtggtgt gacataattg
     3181 gacaaactac ctacagagat ttaaagctct aaggtaaata taaaattttt aagtgtataa
     3241 tgtgttaaac tactgattct aattgtttgt gtattttaga ttccaaccta tggaactgat
     3301 gaatgggagc agtggtggaa tgcctttaat gaggaaaacc tgttttgctc agaagaaatg
     3361 ccatctagtg atgatgaggc tactgctgac tctcaacatt ctactcctcc aaaaaagaag
     3421 agaaaggtag aagaccccaa ggactttcct tcagaattgc taagtttttt gagtcatgct
     3481 gtgtttagta atagaactct tgcttgcttt gctatttaca ccacaaagga aaaagctgca
     3541 ctgctataca agaaaattat ggaaaaatat ttgatgtata gtgccttgac tagagatcat
     3601 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3661 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3721 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3781 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcggatc
     3841 tgcggccacc cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt
     3901 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat
     3961 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
     4021 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
     4081 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     4141 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     4201 gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca
     4261 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     4321 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     4381 cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta
     4441 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     4501 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     4561 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     4621 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     4681 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     4741 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     4801 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     4861 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     4921 ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg
     4981 gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     5041 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5101 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5161 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5221 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5281 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5341 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5401 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5461 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5521 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5581 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5641 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5701 tgctggcctt ttgctcacat gtt
//
